Mapping of Italian research excellence in Neurodegenerative - Apre

Mapping of Italian 

research excellence in 

Neurodegenerative 

Diseases

1. MECHANISMS OF NEURODEGENERATION 

A. Macromolecular interactions and Neurodegeneration 

1. Public health. Therapeutic strategies for the 

prevention and tratment of neurodegenerative 

disease. Molecular basis of amyloid aggregation 

2. Protein conformational transitions that trigger the 

aggregation processes associated to 

neurodegenerative diseases : a nanotechnological 

approach 

Pier Luigi San Biagio 

pierluigi.sanbiagio@pa.ibf.cnr.it 

Bruno Samorì 

bruno.samori@unibo.it 

3. Myelin proteins and demyelinating deseases Eugenia Polverini 

eugenia.polverini@unipr.it 

4. Conformational transitions of proteins from native Maria Grazia Bridelli 

to amyloid form 

mariagrazia.bridelli@unipr.it 

5. Role of altered composition of plasma membrane Sandro Sonnino 

complex lipids in lipid-protein interactions, 

membrane organization and neurodegeneration 

Sandro.sonnino@unimi.it 

6. Alterations of proteolytic pathways in 

Angelo Poletti 

neurodegenerative diseases" 

Angelo.poletti@unimi.it 

7. Optical and biomolecular methods for the 

Antonio Malgaroli 

functional investigation of neural circuits in vivo Antonio.malgaroli@hsr.it 

Gian Giacomo Consalez 

Giacomo.consalez@hsr.it 

8. Molecular mechanisms in neurotransmission: Flavia Valtorta 

effects in neuron and glia phenotypes in synaptic 

efficiency and synapropathy 

Valtorta.flavia@hsr.it 

9. Presynaptic molecular machinery regulationg Maurizio Popoli 

glutamate release an pathophysiological 

mechanism in amyotrophic lateral sclerosis 

Maurizio.popoli@unimi.it 

10. Toward exploiting the screening potential of Daniela Tardito 

miRNome for the identification of genes and 

pathways involved in neurodegenerative disorders 

Daniela.tardito@unimi.it 

11. Identification of signaling pathways in 

Maria Penuto 

neurodegenerative proteinopathies 

Maria .pennuto@iit.it 

12. Neurotensin and neurotensin antagonists in an Tanganelli Sergio 

animal model of Parkinson disease: therapeutic 

perspectives and role of NMDA/neurotensin 

interaction. 

tgs@unife.it 

B. Neurotoxic stimuli 

1. CSF Glycomics in AD and other neurodegenerative 

disease 

2. Intracellular signalling during neurodegenerative 

events triggered upon APP overexpression 

3. Effect of Age and neurodegenerative disorders in 

central neurotrasmission: focus on neurotrasmitter 

relase from neuron and glia 

Domenico Garozzo 

domenico.garozzo@cnr.it 

Maurizio Taglialatela 

m.taglialatela@unimol.it 

Claudio Russo 

claudio.russo@unimol.it 

Mario Marchi 

marchi@pharmatox.unige.it

4. Developing competitive in “ vitro” and “vivo” 

models to study Alzheimer’s disease 

5. Evaluation of mesenchymal stell cells (MSCs) effect 

in Alzheimer’s disease rat models competitive in “ 

vitro” and “vivo” studies 

6. New Tratment strategies (Neurodegeneration and 

Neuroprotection) 

7. Pathways leading to tau pathology – Role of nuclear 

tau in neurodegeneration in fronto-temporal 

dementia 

8. Cholesterol and amyloid-beta: is there a relation? 

- Specific type 4 phosphodiestrerase inhibitors and 

their impact on the production of amyloid-beta 

- Cholesterol and amyloid-beta: is there a relation? 

- Role of non muscle myosin IIB in the processing of 

the amyloid precursor protein (APP) 

9. Basic research 

- Genetic susceptibility to Alzheimer disease and 

genome wide association studies 

- Biobanking 

Francesca Ruberti 

f.ruberti@inmm.cnr.it 

Mariarosaria Miloso 

mariarosaria.miloso@unimib.it 

Patrizia Hrelia 

Patrizia.hrelia@unibo.it 

Fabrizio Tagliavini 

ftagliavini@istituto-besta.it 

Roberta Ricciarelli 

ricciarelli@medicina.unige.it 

Dario Finazzi 

finazzi@med.unibs.it 

10. Neurotoxic stimuli, synaptic dysfunction Calro Sala 

c.sala@in.cnr.it 

11. Neuroprotection by chemokines Cristina Limatola 

Cristina.limatola@uniroma1.it 

12. Phatogenesis and therapeutic targets for SBMA Fabio Benfenati – Mara Pennuto – Chiara 

Scaramuzzino 

Fabio.benfenati@iit.it 

13. Malattia di Alzheimer e trasduzione del segnale Armando Genazzani 

legata al calcio 

14. Mechanisms of neurodegeneration and 

enhancement of adult neurogenesis to improve 

cognitive functions in down syndrome 

15. Pathological mechanisms of neurodegeneration 

triggered by tau and beta-amyloid 

genazzani@pharm.unipmn.it 

Andrea Contestabile 

Andrea.contestabile@iit.it 

Laura Gasparini 

Laura.gasparini@iit.it 

16. Oxidative stress and neurodegeneration Michele Mazzanti 

Michele.mazzanti@unimi.it 

C. Neuro inflammation 

1. Role of vascular inflammation and leukocyte Gabriela Constantin 

trafficking in neurodegenerative diseases 

gabriela.constantin@univr.it 

2. Regional vulmerability of the brain to 

Marina Bentivoglio 

neurodegeneration and immune regulatory 

mechanisms 

marinabentivoglio@univr.it 

3. Neurotoxic stimuli: role of astrocytes and microglia Michela Matteoli 

Michela.matteoli@unimi.it 

4. Gender and hormonal/endocrine signals in Elisabetta Vegeto 

neuroinflammation role in alzheimer’s disease and Elisabetta.vegeto@unimi.it

ain ageing 

5. Role of pro-and anti- inflammatory cytokines in the 

prognosis and progression of neurodegenerative 

diseases 

6. Study of the role of pro-inflammatory chemokines 

in neuroinflammatory disorders and analysis of the 

therapeutic potential of chemokine synthesis 

inhibitors 

7. Role of inflammation and microglial activation in 

neurodegenerative diseases 

8. Role of inflammation in Alzheimer’s disease: study 

of pro-inflammatory cytokines for the identification 

of new pathogenic and diagnostic targets 

D. Oxidative Stress and neurodegeneration 

1. Dopamine and oxidative stress in the pathogenesis 

of Parkinson's disease 

2. Mitochondria, apoptosis and neurodegeneration: 

characterization of mitochondrial dysfunction in 


- Characterization of the molecular mechanisms 

underlying aging, cell senescence and 


3. Structural organization of the mitochondrial 

respiratory chain, generation of reactive oxygen 

species and neurodegeneration 

4. Post-genomic biochemestry of neurodegenerative 

diseases 

Barbara Viviani 

Barbara.viviani@unimi.it 

Claudio Milanese 

c.milanese@angelini.it 

Luisa Minghetti 

luisa.minghetti@iss.it 

Paola Bossù 

p.bossu@hsantalucia.it 

Marco Bisaglia 

marco.bisaglia@unipd.it 

Antonella Bobba 

a.bobba@ibbe.cnr.it 

Giorgio Lenaz 

giorgio.lenaz@unibo.it 

Annalisa Santucci 

santucci@unisi.it 

E. Trophic factors involvement in the neurodegeneration 

process 

F. Endocrine and Metabolic Factors 

1. miRNA as sensors of neuronal stress and 

modulators of synaptic plasticità and memory 

deficits 

2. Developing competitive animal models to study 

Alzheimer’s disease 

3. Genetic susceptibility to Alzheimer’s disease (AD) 

in women: AD as a gender disease 

4. Genetic susceptibility to Alzheimer’s disease 

(Epigenetic maks of susceptibility to Alzheimer 

disese) 

5. Role of altered lipid and cholesterol metabolism in 

neurodegenerations 

Paola Fragapane 

paola.fragapane@uniroma1.it 

Roberto Rimondini 

roberto.rimondini@unibo.it 

Rosa Maria Corbo 

rosamaria.corbo@uniroma1.it 

Lucia Migliore 

l.migliore@geog.unipi.it 

Laura Colombaioni 

laura.colombaioni@in.cnr.it

6. Role of neuroactive steroids in the 

neurodegeneration process 

7. Selective Alzheimer Disease Indicator -1 (Seladinò1): 

a liker between cholesterol, oxidative stress 

and Alzheimer’s disease 

8. Nuclear receptors, coregulators, neuroactive 

steroids and epigenetics in neurodegenerative 

disease associated to metabolic disorders 

G. Cell interactions 

1. The Astrocyte, not just a Bystander in Alzheimer’s 

disease 

2. Synapse and Amyloid-beta: effects on astrocyte 

processes, nerve terminals and network synaptic 

activity 

Roberto Cosimo Melcangi 

Roberto.melcangi@unimi.it 

Roberto Maggi 

Roberto.maggi@unimi.it 

Donatella Caruso 

Donatella.caruso@unimi.it 

Ubaldo Armato 

ubaldo.armato@univr.it 

Manuela Marcoli 

marcoli@pharmatox.unige.it 

3. Alzheimer’s disease, Ageing Luciano Domenici 

domenici@in.cnr.it 

4. Synaptic deficits in neurodegenerative disease Evelina Chieregatti 

Evelina.chieregatti@iit.it 

H. Intracellular Traffiking 

1. Membrane traffic and neurodegenerative diseases: Cecilia Bucci 

role of Rab proteins and their effectors 

cecila.bucci@unisalento.it 

2. Basic research. Neurodegenerative diseases Paolo Macchi 

macchi@science.unitn.it 

I. Animal models 

1. Developing competitive animal models to study Adalberto Merighi 


Adalberto.merighi@unito.it 

2. Synaptic dysfunction in neurodegeneration: Monica DI Luca 

characterization of gene product in functional 

synaptic networks under physiological and 

patological conditions 

monica.diluca@unimi.it 

3. Study of early onset Alzheimer disease in a model Renata Bartesaghi 

for Down syndrome, the Ts65Dn mouse 

renata.bartesaghi@unibo.it 

4. Iron and Neurodegeneration Sonia Levi 

Levi.sonia@hsr.it 

5. Non neurotoxic activity of Beta amyloid in animal Stefano Govoni 

models 

Stefano.govoni@unipv.it 

6. Neurochemical and morphological in vivo and in Tiziana Antonelli 

vitro models in neurodegenerative diseases. ant@unife.it

2. PHARMACOLOGY 

A. Drug Design 

1. Elementary mechanisms of neurodegenerative 

diseases 

2. Innovative drug discovery strategies for Alzheimer’s 

disease 

3. Development of ligands specifically targeting the 

Translocator Protein 18 kDa in Alzheimer disease 

4. Structural studies of acetylcholinesterases and their 

inibitors: implicantions for the design of new antialzheimer 

drugs 

B. Drug screening 

1. Developemt of competitive animal models for AD- 

GLP grade animal facility and behavioural testing 

2. Yeast model to seek for molecules which can 

reduce mitochondrial mutability 

3. Innovative technologies for the therapy of the 

neurodegenerative diseases. 

C. Drug Delivery 

1. Nanotechnology for health: application in the brain 

diseases 

2. Hybrid drug containing reservoirs for long term 

release in brain diseases 

3. New nanodevices targeting beta-amyloid and 

overcoming the blood-brain barrier for the therapy 

of Alzheimer disease 

D. Novel Drugs 

Nino Russo 

nrusso@unical.it 

Carlo Melchiorre 

Carlo.melchiorre@unibo.it 

Federico Da Settimo 

fsettimo@farm.unipi.it 

Alberto Cassetta 

Alberto.cassetta@ts.ic.cnr.it 

Laura Calzà 

laura.calza@unibo.it 

Iliana Ferrero Fortunati 

iferrero@unipr.it 

Luca Ferraro 

frl@unife.it 

MA Vandelli 

Giovanni Tosi 

gtosi@unimore.it 

Angelo Montenero 

angelo.montenero@unipr.it 

Massimo Masserini 

massimo.masserini@unimib.it 

1. Lamberto Maffei 

maffei@in.cnr.it 

2. Synthesis and screening of new AChE/BuChE Aldo Andreani 

inhibitors 

Aldo.andreani@unibo.it 

3. Drug design and synthesis of therapeutic agent for Olga Bruno 

neurodegenerative disorders, particularly for 

Alzheimer disease 

obruno@unige.it 

4. Development of a novel therapeutic strategy for AD Fabrizio Tagliavini 

based on a natural variant of amyloid beta that ftagliavini@istituto-besta.it

hinders amyloidogenesis 

5. Second generation recombinant Abeta peptide 

immunogens as prototype vaccines for AD 

treatment 

Simone Ottonello 

s.ottonello@unipr.it 

6. Pediatric neurodegenerative diseases Maurizio Scarpa 

maurizio.scarpa@unipd.it 

7. Ocular NGF administration as a novel non invasive Paola Tirassa 

approach to protect brain-NGF target neurons that p.tirassa@inmm.cnr.it 

degenerative in Alzheimer’s disease 

8. New symptomatic and neuroprotective terapie for Micaela Morelli 

parkinsons’s disease 

morelli@unica.it 

9. Innovative neuro-reparative strategies via the Maria Pia Abbracchio 

implementation of endogenous neurogenesis and Mariapia.abbracchio@unimi.it 

gliogenesis: focus on the new P2Y-like GPR17 

receptor 

10. Pharmacological modulation of adult neurogenesis Maria Grazia Grilli 

grilli@pharm.unipmn.it 

11. Study of the relationship between the 

Fabrizio Chiti 

structure/morphology of amyloid beta peptide Fabrizio.chiti@unifi.it 

aggregates and their toxicity in cellular, neuronal 

and animal models" 

E. Non-Pharmacological Therapies 

1. Psychoeducational intervention for patients and 

care givers in ParKinson’s Disease and other 


Pio Enrico Ricci Bitti 

pioenrico.riccibitti@unibo.it 

2. Physical excercise and cognitive decline Maurizio Taglialatela 

m.taglialatela@unimol.it 

Alfonso Di Costanzo 

3. New tratment strategies (non pharmacologic 

therapeutic strategies in neurodegenerative 

diseases) 

4. Non invasive brain stimulation an integrated 

approach to neurorehabilitation in Alzheimer 

disease 

5. Early cognitive and electroencephalographic 

markers of normal and pathological ageing 

6. Transplantation of iPS-derived dopaminergic 

neurons for cell replacement in Parkinson's disease 

7. Neural re generation in the cerebellum: 

development of cell replacement strategies for the 

management of spinocerebellar ataxias 

8. Physical activity programs for elderly subjects with 

different grades of cognitive impairment 

9. Promotion of Adapted Motor Activity among 

patients with Parkinson’s disease 

Alfonso.dicostanzo@unimol.it 

Andrea Stracciari 

andrea.stracciari@aosp.bo.it 

Orazio Zanetti 

ozanetti@fatebenefratelli.it 

Patrizia Bisiacchi 

patrizia.bisiacchi@unipd.it 

Vania Broccoli 

Broccoli.vania@hsr.it 

Gian Giacomo Consalez 

Giacomo.comsalez@hsr.it 

Ferdinando Rossi 

Ferdianando.rossi@unito.it 

Federico Schena 

Federico.schena@univr.it 

Enrico Granieri 

Enrico.granieri@unife.it 

patrik.fazio@unife.it 

gino.granieri@unife.it

10. New treatment strategies in Neurodegenerative 

diseases with Deep brain Stimulation 

F. Drug Pharmacogenetics 

1. Role of pharmacogenetics to identify the 

personalized treatment of dementia and 

depression. 

G. Clinical studies 

Mariachiara Sensi 

mchiasen@gmail.com 

Alberto Pilotto 

Alberto.pilotto@operapadrepio.it 

1. Neurocognition in Alzheimer Disease Carlo Caltagirone 

c.caltagirone@hsantalucia.it 

3. HUMAN/CLINICAL RESEARCH 

A. Human –epidemiology/risk factors 

1. Pathogenic mechanisms, early diagnosis and 

prevention of neurodegenerative diseases 

following pesticide intake in animal model 

2. Effects of traffic-related particulate matter on 

transcription of genes implicated in 

neurodegeneration 

Gabbianelli Rosita – Cinzia Nasuti 

rosita.gabbianelli@unicam.it 

Micaela Caserta 

micaela.caserta@uniroma1.it 

3. Cronobiology and neurodegeneration Valerio Carelli 

Valerio.carelli@unibo.it 

chiaralamorgia@gmail.com 

4. Gene-metal interaction as determinant of 

Roberto Lucchini 


lucchini@med.unibs.it 

5. Using prevention to reduce the burden of 

Stefano Mattioli 


6. Construction and development of "Alzheimer-Web- 

GIS". 

Geographical correlation between Alzheimer's 

disease and environmental parameters. Method for 

mapping population-based case-control studies. 

7. Prospective evaluation of biomarkers of 

inflammation in Mild Cognitive Impairment 

subtypes, and their role in the conversion from 

cognitive impairment to dementia and Alzheimer's 

disease 

8. Epidemiology and risk factors for Mild Cognitive 

Impairment, Alzheimer's disease and dementia. 

Population-based 7-year follow-up study. 

s.mattioli@unibo.it 

Giuseppe Mele 

Giuseppe.mele@unisalento.it 

Calogero Caruso 

immunopatologia@unipa.it 

Roberto Monastero 

Roberto.monastero@ki.se 

9. [moved to B.5 Epidemiology genetics] Emilio Di Maria 

Emilio.dimaria@unige.it 

10. Preclinical Diagnosis Carlo Caltagirone 


11. Interactions between the products of the Herpes Manservigi Roberto

simplex genome and Alzheimer's disease 

susceptibility genes 

B. Epidemiology genetics 

1. Genetic susceptibility to Alzheimer’s disease and 

genome wide association studies 

2. Genetic susceptibility to Alzheimer’s disease: 

biobanking 

Roberto.manservigi@unife.it 

Luisa Benussi 

lbenussi@fatebenefratelli.it 

Maria Del Zompo 

delzompo@unica.it 

C. Biomarkers and early diagnosis and imaging 

1. Plasma biomarkers of oxidative stress and 

inflammation in early detection and follow-up of 

dementia. 

2. Vitamins and antioxidants in aging and 


3. Phenotypic and genotypic markers of early 


4. Prioritization and annotation of AD genes, their 

variants and their interaction networks 

5. Cerebrospinal fluid cytokine and chemokine levels 

as potential biomarkers for early diagnosis of 


6. Individual evaluation of cognitive decline: definition 

of low cost non- invasive examination battery and 

implementation of a predictive algorithm to 

identify Mild cognitive impairment 

7. Impiego dei markers biologici e neuro radiologici 

nella diagnosi differenziale precoce 

8. Biomarkers in Mild cognitive Impairment: genetics 

and biochemical and neuroimaging studies for 

early diagnosis. 

Patrizia Mecocci 

mecocci@unipg.it 

Salvatore Monaco 

Salvatore.monaco@univr.it 

Rita Casadio 

casadio@biocomp.unibo.it 

Elio Scarpini 

elio.scarpini@unimi.it 

Paolo Maria Rossini 

Paolomaria.rossini@afar.it 

Orazio Zanetti 

ozanetti@fatebenefratelli.it 

Sandro Sorbi 

sorbi@unifi.it 

9. A multimodality index for early diagnosis of Flavio Nobili 


flaviomariano.nobili@hsanmartino.liguria.it 

10. Lexical semantic skills, APOE and neuroanatomical Paolo Caffarra 

substrate of semantic memory in early diagnosis of 

Alzheimer's disease 

Paolo.caffarra@unipr.it 

11. Cognitive neuroscience of dementia: clinical and Stefano F. Cappa 

theoretical aspect 

Cappa.stefano@hsr.it 

12. Mapping structural and functional brain network Massimo Filippi 

damage in neurodegenerative diseases 

Filippi.massimo@hsr.it 

13. Studies are aimed at investigating whit PET the Daniela Perani 

brain functional parameters and neurotransmission 

systems in neurological and psychiatric diseases 

Daniela.perani@hsr.it 

14. Global proteomics of human fibroblast for the Maurizio Popoli 

identification of early biomarkers for dementia and 


Maurizio.popoli@unimi.it 

15. Parkinson disease and parkinsonisms Gianfranco Spalletta 

g.spalletta@hsantalucia.it 

16. Mild cognitive impairment Gianfranco Spalletta

17. Electrophysiological study of striatal physiology in 

normal conditions and experimental parkinsonism: 

use of genetic and pathogenetic animal models 

18. Celebrospinal fluid biomarkers in alzheimer’s 

disease and other neurodegenerative disease 

D. Diagnosis: biomarkers 

1. Proteomics of cognitive and movement disorders: 

Identification of molecular mechanisms associated 

with disease onset and phenotypic variability of 

frontotemporal lobar degeneration 

2. Peripheral markers of oxidative stress, 

excitotoxicity and abnormal protein metabolism in 

neurodegenerative disorders. 

3. Validation and search of CSF biomarkers for the 

early differential diagnosis of cognitive impairment. 

Development of new diagnostic approaches for 

prion diseases in humans. 


Paolo Calabresi 

calabre@unipg.it 

Lucilla Parnetti 

parnetti@unipg.it 

Roberta Ghidoni 

rghidoni@fatebenefratelli.it 

Carlo Ferrarese 

Carlo.ferrarese@unimib.it 

Piero Parchi 

Piero.parchi@unibo.it 

4. Analisi Biochimica di parametri di stress ossidativo Gabriele Siciliano 

gsicilia@neuro.med.unipi.it 

5. Cognitive decline in multiple sclerosis Laura Calzà 

6. Search of predictive markers of AD based on 

amplification of disease-specific amyloid-beta 

oligomers 

Laura.calza@unibo.it 


ftagliavini@istituto-besta.it 

7. Mild cognitive impairment and dementias Gianfranco Spalletta 


E. Diagnosis: neuropsychology 

1. Handwriting analysis as a novel diagnostic tool for 

Alzheimer’s disease in the clinical and forensic 

settings 

2. Amnesic Mild Cognitive Impairment: 

neuropsychological features and new diagnostic 

criteria (Dubois et Al., 2007) 

Maurizio Balestrino 

mbalestrino@neurologia.unige.it 

Angiola Maria Fasanaro 

Giola.fasanaro@virgilio.it 

3. Predictors of conversion from MCI to dementia Roberto Gallassi 

Roberto.gallassi@unibo.it 

4. Individual evaluation of cognitive decline; validation Ildebrando Appollonio 

of ultrafast screening tools. 

ildebrando.appollonio@unimib.it 

F. Biobanking Bioinformatics 

G. Ambient assisted living and socioeconomics 

1. Home-based Empowered Living for people with 

dementia and Parkinson’s disease 

Roberto Monastero 

roberto.monastero@unipa.it

2. Neuro-rehabilitation and brain functional imaging: 

rehabilitation customization driven by best 

exploitation of celebral plasticity and health system 

sustability. 

3. Role of information and communication technology 

(ICT) systems to improve quality of life, quality of 

care and safety of older patients with cognitive 

decline 

4. A transdisciplinary orented proposal from a social 

sciences perspective 

Alessandra Pedrocchi 

pedrocchi@biomed.polimi.it 

Alberto Pilotto 

alberto.pilotto@operapadrepio.it 

Vincenzo Maria Bruno Giorgino 

Vincenzo.giorgino@unito.it


A- Macromolecular interactions and 

Neurodegeneration

Nome Dr. Pier Luigi San Biagio 

Contatti e-mail: pierluigi.sanbiagio@pa.ibf.cnr.it 

phone: +39 091 6809311 

cell: +39 329 410 5564 

Istituto/Dipartimento Istituto di Biofisica - CNR, Via Ugo La Malfa, 153 , 90146 Palermo (Italy) 

Proposta di ricerca Many untreatable neurodegenerative human disorders are associated with the 

aggregation of misfolded or natively unfolded proteins into amyloid fibrils. Our 

research activity concerns Alzheimer’s Parkinson’s and Albers-Schönberg diseases 

with the shared perspective of relating the understanding of the pathway leading 

to the fibrils formation to the design of appropriate drugs capable on preventing 

the protein aggregation. 

Alzheimer’s disease. Recent results suggested that the main neurotoxic species in 

Alzheimer’s Disease (AD) would not be the 

insoluble protein aggregates, made of extracellular deposits of Beta-amyloid 

peptide (Ab) fibrils, but rather the soluble oligomeric 

species, including small globular structures, 2.7 to 6.0 nm in diameter, and 

“protofibrils”. Conformational constrains imposed by small 

organic molecules could play a key role in modulating the fibrillogenesis process, 

so providing effective therapeutic tools to target both 

oligomeric and fibrillar species. In this perspective, polycyclic aromatic molecules 

could be of special interest as they might disrupt the 

molecular architectures precursors of fibrils by means of aromatic interactions, 

like stacking interactions with tyrosine and 

phenylalanine residues of Ab. In particular, natural polyphenols, composed of one 

or more aromatic phenolic rings, are a class of 

phytochemicals found in high concentrations in wine, tea, nuts, berries, fruits, 

cocoa, and an ample assortment of other plants. Some of 

them have been demonstrated to have anti-oxidant and anti-amyloidogenic 

effect and it has been speculated that they could prevent 

the development of Alzheimer's disease, not only through scavenging of reactive 

oxygen species, but also through directly inhibiting 

the deposition of fibrillar beta-amyloid in the brain. In the present project, we’ll 

test some natural molecules, such as ferulic acid (FA), a 

phenolic anti-oxidant present in fruit cell walls, hypericin (Hyp), a natural pigment 

extracted from Hypericum perforatum, widely used as 

a mild antidepressant and epigallocatechin gallate (EGCG), one of the major 

flavonoids of green tea. The aggregation process of Ab in 

the presence of these natural molecules will be studied by light scattering, 

steady-state and time-resolved fluorescence, FTIR and CD 

techniques. The analysis of the correlations between the effects of the studied 

compounds on the various stages of amyloid fibril 

formation, and their known physicochemical properties, will provide novel 

insights on the specific role of hydrophobic and aromatic 

interactions. Encouraging results have already been obtained using hypericin 

which has been shown to affect and to interfere with the 

early stages of polymerization process. The furtherance of this type of study, 

extended to other small molecules like FA and EGCG, 

hopefully will meet the goal of designing effective therapeutic tools to ideally 

target both oligomeric and fibrillar species, thus 

contributing to devise AD prevention strategies.

Area di interesse 

identificata 

Finanziamenti ricevuti 

Titolo progetto 

Ente finanziatore 

Durata progetto 24 months, started 2001 

Parkinson’s disease. Alpha-synuclein (aS) is the main constituent of Lewy bodies, 

intraneuronal fibrillar inclusions which are present 

in all patients affected by Parkinson’s disease (PD). From genetic and biochemical 

studies it is believed that aS is involved in the 

disease. AS is a natively unfolded protein with the ability to acquire secondary 

structure upon interaction with membranes or with itself. 

It is linked to PD by two evidences: the accumulation of amyloid fibrils of the 

protein and the autosomal dominant forms of the disease 

(A53T, A30P and E46K mutants). Both the biological role of this protein and the 

mechanisms involved in the ethiopathogenesis of PD 

are still unknown. The protein is located at the presynaptic terminal of neurons in 

all the CNS, where it exists free in the citosol or 

bound to synaptic vesicles. The membrane binding causes the formation of an 

amphipatic alpha-helix in the first part of the protein, which lies at the membrane 

surface without crossing the bilayer. Recent evidences show that aS is able to 

bind and permeabilize cell 

membrane. Our investigation will be devoted (i) to the characterization, at the 

single molecule level, of the channel like ability of aS and its architecture. The 

pore-forming ability of monomers and aggregates of different sizes will be 

investigated. Secondly, the lipid specificity will be also investigated by means of 

high resolution NMR and MS (both maldi-tof and esi-ms). The ability of aS to 

provoke or protect unsaturated acyl chains of membrane lipids from oxidation 

will also be investigated, by studying with MS the products of oxidation. As 

possible therapeutic perspectives, the organic or peptidic inhibitors of proteinprotein 

interaction or able to clog the already formed pores will be investigated. 

Furthermore, if indications of any lipid specificity will be obtained, we will 

develop lipid formulates suitable for sequestering aS lowering its level in solution. 

Albers-Schönberg disease. Mutations in the gene coding for the lysosomal Cl-/H+ 

exchanger CLC-7 lead to recessive osteopetrosis 

and Albers-Schönberg disease. Apart from the osteopetrotic phenotype the 

recessive disease is characterized by severe generalized 

neurodegeneration. Also knock-out of the plasma membrane localized CLC-2 lead 

to neurodegeneration (retinal degeneration and 

leukoencephalopathy). The mechanisms underlying these phenotypes are 

unclear. We are studying the functional properties of these 

two proteins using electrophysiological and optical methods, combined with 

mutagenesis, in order understand how their dysfunction 

may lead to neurodegeneration. In particular, we consider these proteins as 

potential risk factors for more generalized forms of 

neurodegeneration. 

Public health. Therapeutic strategies for the prevention and treatment of 

neurodegenerative 

diseases. Molecular basis of amyloid aggregation 

Progetto strategico di ricerca finalizzata sulla malattia di Alzheimer (Art. 12/bis D.Lgs 

229/99), 

“Beta-amyloid deposit on the cell membrane: role of metal ions and free radicals” 

Italian 'Ministero della Salute”

Abstract del progetto 



Durata progetto 


















Characterization of some key molecular mechanisms for the neuronal damage connected 

to AD. Specific objectives: (i) to identify of the key steps responsible for the amyloid 

depost formation on membrane model systems in vitro; (ii) to evaluate the role of the 

membrane lipid composition on the deposit; (iii) to understand the role of metal ions and 

oxidative stress on 

the fibril growth both in solution and on membranes; (iv) to establish an in vitro test for 

evaluating the ability of drugs to inhibit bA aggregation. 

“Animal neuropathies: molecular and functional analysis of the prionic proteins in Sicilian 

bovine race ” 

Italian 'Ministero della Salute 

24 months, started 2003 

The main objective of the present proposal is to understand the mechanisms by which 

wild type and mutant prion proteins become misfolded in such a way as to provoke 

neurodegenerative diseases becoming transmissible agents. An extensive study of the 

aggregation process will be done by using static and dynamic light scattering and 

fluorescence techniques. 

Prin 2005 project: “The effects of metal ions on the protein aggregation processes” 

Italian 'Ministero Università e Ricerca' 


The project concerns the study of the effects of metal ions in solution on the protein 

aggregation processes. Main goals are (i) to understand the molecular mechanisms 

relevant for amyloid aggregation and to establish the relevance of metals in modulating 

solvent mediated protein-protein interactions. 

“Electrophysiological characterization of the alpha synuclein channels, a protein involved 

in Parkinson’s disease” 

Local Bank Foundation CARITRO 


The aim of the project is: i) to characterize the channel like activity of alpha synuclein (aS) 

and its role in the disease; ii) to distinguish the most active aS form; iii) to verify the ability 

of aS to bind to membranes and lipid requirements; iv) to investigate the role of aS in the 

prevention of the oxidation of unsaturated fatty acid chains 

Prin 2008 project: “Inhibition of beta-amyloid peptide aggregation by some natural 

compounds” 

Italian 'Ministero Università e Ricerca 


The aim of this project is to provide a molecular approach for preventing Abeta 

aggregation and toxicity through the use of some natural molecules potentially able of 

interact with Abeta by stabilizing or disrupt aggregates of different sizes. 

“Membrane transport of chloride and protons: mechanism of function of CLC family 

proteins involved in epilepsy and neurodegeneration” 

Compagnia San Paolo 


CLC-2 is a plasma membrane localized Cl- channel whereas CLC-7 is a lysosomal Cl-/H+ 

exchanger. Knock-out of CLC-2 in mice leads to retinal degeneration and 

leukoencephalopathy and knock-out of CLC-7 and human mutations in the CLCN7 gene

lead 

to generalized neurodegeneration. The aim of the project is to understand better the 

transport 

function of these two proteins in order to get insight into their causative role in these 

forms of 

neurodegeneration.

Nome Bruno Samorì 

Contatti E mail: bruno.samori@unibo.it 

Istituto/Dipartimento Laboratory of Nanoscience and Nanobiotechnology, Department of 

Biochemistry, University of Bologna (Italy) 

(http://biocfarm.unibo.it/samori/) 

Proposta di ricerca 

Area di interesse identificata Basic research 

Abstract According to an emerging view, the aggregation-prone proteins associated 

to neurodegenerative, Parkinson’s, Alzheimer’s, and Creutzfeldt-Jakob 

diseases, can switch back and forth between functional and 

amyloidogenic/prionogenic conformations. Their conformational 

polymorphism poses tremendous challenges to standard methods in 

structural biology, which mask the complexity of their behaviour by 

recording observables as time and ensemble averages. Single-molecule 

methodologies can instead follow the time trajectories of individual 

molecules or molecular assemblies, can map their energy landscapes, and 

obtain distributions of molecular properties. These capabilities can provide 

information about the monomeric state of an amyloidogenic protein and 

about the very early stages of its amyloid cascade. This information is 

crucially needed for the design of new drugs against Parkinson’s or 

Alzheimer’s disease that must be capable to target upstream of the 

formation of the most neurotoxic oligomeric species. In fact concerns are 

now growing over the possibility that more harm than cure can be created 

whenever the deposits in the central−nervous−system, are targeted 

instead: in this way their dynamic equilibrium with the oligomeric forms 

can be shifted back to the latter. 

We have recently shown that Atomic Force Microscopy-based Single 

Molecule Force Spectroscopy (AFM-SMFS) can single out the different 

monomeric conformers of α-synuclein (the intrinsically unstructured 

protein associated to Parkinson’s disease) and of its mutants associated to 

genetic PD. (1,2). So far AFM-SMFS has been the only methodology that 

made it possible to achieve this goal. 

Ongoing projects: 

a) The integration of AFM-SMFS with Optical tweezers (OT) (in coll. 

with Carlos Bustamante (UC Berkeley). 

b) The determination of alpha synuclein aggregation step targeted 

by different potential drugs or chaperons, by using an integrated 

approach based on AFM- and OT-SMFS, AFM-imaging, 

Fluorescence Polarization Spectroscopy ( in collaboration with



Luigi Bubacco, University of Padova) 

c) The conformational equilibria or prion proteins (in coll. with G. 

Legname SISSA Trieste, and Motomasa Tanaka, Riken, Japan) 

(1) Sandal, M., et al (2008). PLoS Biol 6, 999-1008. 

(2) Brucale, et al. (2009). Chembiochem 10, 176-83. 

Nanomanipolazione di singole molecole delle proteine coinvolte in 

malattie neurodegenerative 

Ente finanziatore FIRB Nanobiotecnologie per dispositivi e sensori innovativi (G.U. 

29.07.2005 n. 175) 

Durata progetto 2005-2008; 2005-2010 

FIRB Metodologie e tecnologie innovative per la farmaceutica. CINECA 

RBNE03PX83 

Abstract del progetto Sviluppo metodologie di nanomanipolazione di proteine intrinsecamente 

non foldate o disordinate 

Titolo progetto Meccanismi Neurodegenerativi nelle Malattie da Prioni (coordin. Catia 

Sorgato) 

Ente finanziatore PRIN 2009 

Durata progetto 2010-2012

Nome Dr. Eugenia Polverini 

Contatti 

eugenia.polverini@unipr.it 

Istituto/Dipartimento Department of Physics – University of Parma 


Our studies are - and will be - focused on myelin proteins, to the aim of clarify their structure to function relationship 

and the mechanisms involved in demyelinating diseases, including multiple sclerosis. Two main proteins are under 

investigation: the Myelin Basic Protein (MBP) and the P2 myelin protein. 

MBP is a highly post-translationally modified structural component of central nervous system myelin, the 

multilamellar membrane wrapped around nerve axons. MBP has a high net positive charge and its principal role 

seems to be the adhesion of the cytosolic surfaces of the compact myelin sheath, whose structural integrity 

determines the speed of transmission of action potentials along the axon. However, MBP seems to be a 

multifunctional protein, since it also interacts with other cytoskeletal and signalling proteins, tethering some of them 

to the oligodendrocyte plasma membrane. The three-dimensional structure of MBP in situ is still unknown, due to its 

intrinsic flexibility and dependence of conformation on environment. Therefore a neighbourhood-dependent 

structural charcterization of fragtments is attempted. In the classic 18.5 kDa MBP sequence, there is a highly 

conserved fragment, constituting two regions with different putative functionality. The first region seems to 

associate with the membrane, and comprises the primary immunodominant epitope in multiple sclerosis; the second 

one is a proline-rich segment, that has been predicted to be a ligand for protein kinase SH3-domains. Due to the 

functional relevance of this fragment of MBP, we’re trying a structural characterization - in the absence and in the 

presence of protein’s post-translational modifications - by means of molecular dynamics simulations in membrane 

and of spectroscopic techniques (in collaboration with the University of Guelph, Ontario, Canada). 

P2 myelin protein is a basic peripheral nervous system protein localized to the cytoplasmic spaces in the myelin 

lamellae. It was recognized as the factor inducing the Experimental Allergic Neuritis, an animal model for the study of 

the Guillain-Barrè syndrome, a demyelinating disease of the PNS in humans. The interest for the putative 

autoantigenic role of the P2 protein and the need to understand its function in the myelin sheath has led to the study 

of its structural characteristics and behaviour, basing on the solved crystal structures. By investigating its interaction 

with different kinds of lipids, on the one hand we want to clarify its transport role and on the other hand we want to 

verify its putative involvement in the compactness of PNS myelin sheat. Different approaches are carrying out: 

molecular docking simulations, spectroscopical investigations and, in collaboration with the University of Oulu, 

Finland, crystal X-ray diffraction and small-angle X-ray scattering. 

Area di interesse identificata 






myelin proteins and demyelinating deseases 

University local research funds (FIL)

Nome Prof. ROBERTO DE RENZI - Dr. MARIA GRAZIA BRIDELLI 

Contatti 

mariagrazia.bridelli@unipr.it 

Istituto/Dipartimento Physics department 

University of Parma 

Parco Area delle Scienze, 7/A, Parma 

ITALY 


The goal of our research is threefold: 

1) study of the nucleation rate and the growth rate of the amyloidogenic process by controlling and modulating the 

solution parameters such as temperature, pH, solvent composition, and peptide concentration, not only influencing 

the final fibril state but also the formation kinetics. The analysis of the aggregation process will be performed in 

solution by employing CD and Fluorescence spectroscopies by following the change of fluorescence emission of small 

hydrophobic molecules, called amyloid ligands, which do not bind to specific proteins but are selective for protein 

aggregates with a cross β-sheet conformation. Dynamic Light Scattering measurements will allow to monitor size and 

shapes of the fibrils during the aggregation process. 

2) study the conformational state of proteins in the native and amyloid form at very low hydration level by 

investigating the structural properties of the first hydration shell. It is well known that proteins are surrounded in the 

cell by a hydration shell formed by interacting water molecules that form patches locally distributed in dependence 

on the heterogeneous surface chemistry or occluded in the internal cavities. This bound water in the crowded 

cytoplasm may not be enough to fully cover the protein surface with a monolayer aqueous sheath, moreover about 

0.4 g of water/g of protein are enough to assure protein normal functionality. Two experimental techniques can be 

applied to investigate this structured water to gain information on the geometry of the protein surface and 

secondary structure. Thermally Stimulated Depolarization Currents (TSDC) technique, applied in the past to a variety 

of natural and synthetic biopolymers has recently been applied to the study of lysozyme. It has allowed recognizing 

the main features of the hydration as a function of secondary structure, revealing a clear dominance of the βstructure 

in the amyloid form. Fourier Transform Infrared Spectrometry as well has proven to be highly versatile in 

the analysis of protein conformation by monitoring the alteration in protein secondary structure as changes in the 

Amide bands. 

3) by means of the molecular dynamics simulation techniques, study the conformational changes that proteins 

undergo under the critical environmental conditions that in vitro cause the fibrils formation. This approach could be 

useful to investigate the pathway(s) of the transition and its molecular features. 


Finanziamenti ricevuti University local research funds FIL 2008 

Conformational transitions of proteins from native to amyloid form. 

Titolo progetto Hydration structure analysis of Lysozyme amyloid fibrils 

Ente finanziatore Ministero dell’Istruzione, dell’Università e della Ricerca 

Durata progetto 2 y 


Combined Thermally stimulated depolarization currents (TSDC) and FTIR 

techniques have been employed to investigate the conformation of hen egg 

white lysozyme in native and amyloid form, in the state of powder at very low 

hydration level. The spectra reveal specific features that can be attributed to 

water texture around the secondary structure adopted by the macromolecule.

Nome Sandro Sonnino 

Contatti 

Tel. 

E mail 

Via fratelli Cervi 93, 20090 Segrate (Mi) Italy, 

+39 0250330360, fax +39 0250330365, 

sandro.sonnino@unimi.it 

Dipartimento Medical Chemistry, Biochemistry and Biotechnology, University of Milan 


Role of altered composition of plasma membrane complex lipids in lipid-protein interactions, membrane 

organization and neurodegeneration. 

Area di interesse identificata 1. MECHANISMS OF NEURODEGENERATION 

F. Endocrine and Metabolic Factors

Nome Angelo Poletti 

Contatti 

02-5031.8215 

angelo.poletti@unimi.it 

Istituto/Dipartimento Dipartimento di Endocrinologia, Fisiopatologia e Biologia Applicata , Centro di 

Eccellenza sulle Malattie Neurodegenerative Universita' degli Studi di Milano 

via Balzaretti 9 - 20133 Milano 


Area di interesse identificata Macromolecular interactions and Neurodegeneration 





ROLE OF ALTERED PROTEOLYSIS IN THE DEGENERATION OF MOTOR NEURONS IN 

in vivo AND in vitro MODELS OF FAMILIAL AMYOTROPHIC LATERAL SCLEROSIS. 

INVOLVEMENT OF CONSTITUTIVE AND IMMUNO PROTEASOMES AND OF THE 

AUTOPHAGY−LYSOSOME PATHWAY 

TELETHON – Italy 

three years (end 2010) 


Amyotrophic lateral sclerosis (ALS) is a neurological disorder primarily affecting 

motor neurons, i.e. the neurons responsible for the contraction of muscles that 

control voluntary movements, speach, swallowing, respiration. The disease, 

whose causes are still largely unknown, is characterized by a progressive 

paralysis that leads to death for respiratory failure. As in some familiar forms of 

ALS there are mutations in the gene 

that codes for the enzyme superoxide dismutase 1 (SOD1), it has been possible 

to create experimental models of the disease to facilitate the research on the 

causes that lead to motor neurons death. Both in ALS patients and in the 

experimental models the vulnerable neurons contain aggregates of abnormal 

proteins, whose accumulation can be responsible for the degeneration. It is 

therefore possible to hypothesize that a role in the pathology is played by 

dysfunctions of the proteolytic mechanisms, i.e. of the mechanisms that normally 

degrade intracellular protein. We propose to investigate whether motor neurons 

degeneration in ALS is due to a failure of protein degradation, using two 

experimental models (transgenic mice and a motor neuron cell line, both 

expressing mutant human SOD1) and analyzing two different intracellular 

proteolytic systems: the proteasomes and the autophagosome−lysosome 


Motorneuron degeneration in Spinal and Bulbar Muscular Atrophy: from the 

molecular mechanisms to the potential therapeutical approaches 

Ente finanziatore TELETHON - ITALY 


Three Years (End 2011) 


CAG/polyglutamine (polyQ) diseases, the largest class of hereditary 

neurodegenerative diseases, include SpinoBulbar Muscular Atrophy (SBMA), a 

motorneuron disease linked to the androgen receptor (AR) mutation. SBMA is a 

valuable model of polyQ diseases, because ARpolyQ neurotoxicity is triggered by 

testosterone, the endogenous AR ligand, possibly via protein conformational 

changes. In SBMA animal models, the symptoms are reversed by testosterone 

removal suggesting that SBMA patients can be treated with drugs preventing 

testosterone action. The overall objectives of the study are to find the 

mechanism(s) by which ARpolyQ neurotoxicity can be modulated by 

testosterone with the aim to identify novel therapeutical approaches for SBMA 

and possibly indications useful for other polyQ diseases. Therefore, all the 

studies proposed will be carried out in absence or in presence of testosterone, 

and four different specific aspects, thought to be involved in SBMA, have been 

selected: 

− interactions of the two major intracellular degradative systems: the


ubiquitin−proteasome and the autophago−lysosome pathways in the removal of 

the neurotoxic ARpolyQ; analysis of ARpolyQ solubility and aggregation, and their 

modulation by selective AR modulators (SARM) and HSP−modulators. 

− analysis of the axonal antero−retrograde transport in presence of ARpolyQ 

activated by testosterone (and SARMs). This aim will include the expression 

analysis of some proteins related to axonal outgrowth and controlled by 

testosterone (neuritin or CGP15). 

− analysis of the possible toxicity of ARpolyQ in muscle cells, and the interaction 

between motorneuron and muscle cells in SBMA. 

− analysis of the AR promoter activity to identified factors downregulating the 

ARpolyQ levels in motor neuron. These studies will provide information of the 

mechanisms by which ARpolyQ becomes neurotoxic after testosterone 

interaction, that may be used to design novel therapeutical approaches for SBMA 

and related diseases 

Alterations of Axonal Functions and Neurodegeneration in Motor Neuron 

Diseases 

Ente finanziatore FONDAZIONE CARIPLO 

Durata progetto Three Years (end 2012) 


Motor neuron diseases (MNDs) are a group of sporadic or hereditary 

neurodegenerative diseases characterized by selective motor neuron death. They 

may affect children and adult patients with relentless progression rate leading to 

amyotrophic paralysis and death. Several genetic, molecular and cellular 

alterations have been reported in the different MNDs and interpreted as 

potentially relevant in the pathogenesis responsible for neuronal death. 

However, it is still unclear whether these alterations are cause or effects of initial 

insults capable to perturb cell functions. The clinical trials attempted with the 

aim to counteract these alterations have proven to be largely ineffective. A 

common aspect of most MNDs is the alteration of motor neuron axons, thus 

suggesting specific axonal function impairment. Axonal processes can reach a 

considerable length (up to 1 meter) and are fundamental for motor neuron 

functions and survival. Axons allow neuronal control of muscle contraction, but 

also collect survival/trophic factors released from the muscle, and required for 

motor neuron physiology. Nerve damages and axonal loss result in motor neuron 

death and muscle atrophy. The project will focus on three major human MNDs, 

spinal muscular atrophy (SMA), spino-bulbar muscular atrophy (SBMA) and 

amyotrophic lateral sclerosis (ALS), affecting similar motor neuron population 

with different rate of cell death. It will be based on advanced analytical 

techniques and specific expertise of five research units. The units will utilize 

complementary genomic, biochemical, molecular and cellular assays to study the 

mechanisms of axonal dysfunctions in the MNDs studied. Since gene mutations, 

gene expression impairment, and altered post-translational modifications of 

specific proteins are involved in these MNDs, the project will be dedicated to 

unravel such alterations at the genomic and molecular level with the final aim of 

identifying novel common therapeutical targets. Main objectives: i) 

development and/or improvement of motor neuron models of SMA, SBMA and 

ALS; ii) analysis at the (nuclear and mitochondrial) genomic level of alterations 

determined by the lack or the expression of proteins (FL-SMN/a-SMN, mutant 

ARpolyQ, mutant SOD1) responsible for the MNDs selected; iii) morphological, 

cellular, molecular and biochemical analyses of the effects of the considered 

proteins on axonal functions/dysfunctions in the MNDs selected.Five different 

research institutions (University of Milan, Center of Excellence on 

Neurodegenerative Diseases; IRCCS Foundation Neurological Institute C. Besta; 

IRCCS Italian Auxologic Institute; Mario Negri Institute for Pharmacological 

Research; National Institute of Molecular Genetics) with specific expertise in the 

basic mechanisms of neurodegeneration will be involved. The approval of this 

project would enable to establish a critical mass of researchers devoted to MNDs 

in Milan, allowing the recruitment and formation of young scientists.

Nome Antonio Malgaroli, Gian Giacomo Consalez 

Contatti 

02 2643 4886 

antonio.malgaroli@hsr.it, giacomo.consalez@hsr.it 

Istituto/Dipartimento Università Vita-Salute San Raffaele 

Proposta di ricerca Optical And Biomolecular Methods For The Functional Investigation Of Neural Circuits In Vivo. 

Area di interesse identificata Analisi della funzione sinaptica nei tessuti nervosi normali e in modelli di 

neurodegenerazione 





Optical And Biomolecular Methods For The Functional Investigation Of Neural 

Circuits In Vivo. 

Fondazione CARIPLO (pending) 

2 anni 

Every human behavior depends on an activity change in one or more brain areas, 

often with the contribution of very small subgroups of cells and synapses. 

Conversely, when a brain disease whatsoever, a tumor, a vascular disorder, a 

grey or white matter pathology, etc., modifies some behavioral aspect, inevitably 

this effect arises from a change in neural activity. Hence, viewing the fine 

structure of the brain by the most advanced imaging techniques is certainly 

important but it would be more important to find some effective way to detect 

changes in neuronal and synaptic activity during both physiological and 

pathological processes. Indeed, for many different body organs, the kidney, the 

heart, the liver, the pancreas, etc, the early detection of signs and symptomsand 

the choice of an optimized therapeutic approach often depend more on the 

availability of a large set of chemical and molecular markers of cell function 

rather than on our ability to visualize fine details of the tissue anatomy. 

Unfortunately techniques available today to monitor brain function, such as 

E.E.G., fMRI, fNIRS, etc., either sample brain activity very indirectly, through 

changes in metabolism and blood flow, or do not have the resolution required to 

pinpoint changes in small networks of neuronal cells and synapses. At present, 

anomalous states of brain activity can be detected only when they arise from 

the concerted activity of large numbers of neurons and synapses, such as in 

epilepsy, or when cell and/or synaptic and/or axonal damage becomes very 

profound. For all the above reasons, the demand for a methodology that could 

yield a more direct readout of in vivo neuronal and synaptic activity is extremely 

high. 

In this research project we are planning to further develop a methodology that 

was recently designed in one of our labs to provide a very direct and sensitive 

readout of brain activity. This is based on a family of recombinant molecules 

which were designed in our labs to detect synaptic activaton via the application 

of exogenous markers. This methodology is capable of revealing the physiological 

activation of brain synaptic networks with very high sensitivity and resolution. 

These striking results have prompted the generation of a series of transgenic

animal models and analytical tools that will be used for further in vivo validation 

and for the selective probing of specific subgroups of neuronal cells and 

pathways, both in physiological and pathological conditions. The final goal of this 

project is to test the applicability of this approach to the quantitative analysis of 

brain activity. We expect that this work will generate important information for 

the development of similar approaches to the human brain.

Nome Flavia Valtorta 

Contatti 

Tel. 

E mail 

Flavia Valtorta, Fabio Grohovaz, Daniele Zacchetti 

02-2643-4826 (Valtorta) 4811 (Grohovaz), 4817 (Zacchetti) 

valtorta.flavia@hsr.it, grohovaz.fabio@hsr.it, zacchetti.daniele@hsr.it 

Istituto/Dipartimento San Raffaele Scientific Institute, Division of Neuroscience 

Via Olgettina 58 

20132 Milano, ITALT 

Proposta di ricerca. 

Molecular mechanisms in neurotrasmission: effects of neuron and glia phenotypes in synaptic efficiency and 

synaptopathy 

Comprehensive goal of our research is the elucidation of the molecular mechanisms that underlie communications 

between cells of the brain, both in the adult and during development. This proposal involves the complementary 

expertise and the tight collaboration of two research units: the NEUROPSYCHOPHARMACOLOGY UNIT (Flavia 

Valtorta) and the CELLULAR NEUROPHYSIOLOGY UNIT (Fabio Grohovaz and Daniele Zacchetti). 

The main form of communication between neurons is achieved at the synaptic level through regulated secretion of 

neurotransmitter. This process is of paramount importance for information processing in the central nervous system 

and is known to undergo dysfunction in a number of pathophysiological states. Substantial advancements have been 

made concerning the identification of the proteins involved in the basic mechanism of neurotransmitter release. 

Much less is known about the molecules that participate to the assembly of the complex synaptic machinery during 

development. 

Research in Valtorta’s group will develop along two main lines: Aim 1: Diseases of synaptic origin. The main focus will 

be on the family of the synapsins and their link to epilepsy and autism. Synapsins are a family of brain 

phosphoproteins involved in neurotransmitter release through regulation of synaptic vesicle availability. Recent data 

from the lab show that synapsins have also an important function in the organization of synaptic contacts. Synapsin 

KO mice develop an epileptic phenotype, and mutations in the human synapsin genes have been recently associated 

with epilepsy and autism spectrum disorders. The central interest is to investigate the molecular roles of synapsin in 

synapse formation and function, and how these roles relate to the development of brain pathology. Another project 

concerns the identification of novel molecular interactors of synuclein and of their role in neurodegeneration 

associated with Parkinson disease. 

Aim 2: Membrane trafficking in neuronal development. The process that leads from undifferentiated, round neuronal 

precursors to fully differentiated, highly polarized neurons is extremely complex. Among the various phenomena 

driving this process, those associated with membrane trafficking are of fundamental importance. The group is 

interested in studying membrane trafficking in the very early steps of neuronal development. In this respect, they 

have identified a process of developmentally-regulated, high-capacity endocytosis as one of the earliest marker of 

neuronal polarity, and will investigate its function in neuronal commitment and axon pathfinding. This investigation 

will be extended to the phenotypical characterization of the early steps of neuronal commitment of neural 

stem/progenitor cells and iPS (induced-pluripotent stem cells). In the same cells, the late steps of neuronal 

development will also be studied, i.e. the formation of synapses and their functional refinement, which are essential 

phenomena for the proper integration of cells in neuronal networks. 

The work in Grohovaz’s research unit is focused on the general comprehension of the physiopathological 

mechanisms underlying neuroprotection or leading to neurodegeneration. The group mostly uses brain primary 

cultures and applys molecular biology and in vitro cellular approaches, including live cell imaging (calcium imaging 

and total internal reflection microscopy). Electrophysiology and animal behavioral studies are conducted in 

collaboration with other research groups. 

A main project deals with the generation of oxidative stress and the ensuing protection mechanisms in both neurons 

and astrocytes. In particular, the mechanisms of iron entry and its influence on the production of reactive oxygen 

species are investigated. On the other hand, in light of the importance of neuroinflammation, interest is also directed 

to the role of glia in the neurodegenerative processes. More specifically, the molecular mechanisms involved in the 

process of glia activation and the influence the activated phenotype has on neuronal survival are investigated. 

Along with the study of these general physiopathological mechanisms, the group addresses specific issues within the 

following disease-oriented projects: 

(i) Alzheimer’s disease pathogenesis: the regulation of BACE1/BACE2 expression; the implication of beta-secretase 

activity on amyloid-beta deposition; the cellular effects of the various amyloid-beta forms on different cell types from 

brain. 

(ii) Derangement in intracellular iron homeostasis and hereditary ferritinopathy: analysis of the mechanisms of iron

entry; cytotoxic effects of iron overload; cellular effects of ferritin light chain mutations. 

(iii) Pathogenesis of neuronopathic (type A) Niemann-Pick disease: role of sphingosylphosphocholine on changes in 

astrocytic phenotype; impairment of neuronal function. 

Area di interesse identificata Synaptic disfunction and neuron-glia interplay 


Titolo progetto Research unit in Molecular Neuroscience 

Ente finanziatore I.I.T. (Istituto Italiano di Tecnologia) 

Durata progetto 5 years (2008-2012); 600'000 EUR 


Neurotransmitter release from nerve endings is the main form of information 

transfer from one neuron to another or to an effector cell. Release occurs by 

exocytosis from storage organelles, the synaptic vesicles, and is triggered by the 

action potential-induced depolarization of the nerve terminal. The project pivots 

on the idea that synapses perform an electro-chemical computation of both local 

and environmental signals they receive. In this respect, a key role is played by 

astrocytes, the close partners of neurons that, upon stimulation, release a 

number of neurotransmitters and signaling molecules. The development of these 

processes is investigated by developing and exploiting innovative molecular 

imaging approaches.

Nome Maurizio Popoli 

Contatti 

Tel. 

E mail 

Tel: +390250318361 

E-mail: maurizio.popoli@unimi.it 

Istituto/Dipartimento Center of Neuropharmacology, Department of Pharmacological Sciences, 

CEND, University of Milan 


Area di interesse identificata 1A. Presynaptic molecular machinery regulating glutamate release as 

pathophysiological mechanism in amyotrophic lateral sclerosis 





Abstract del progetto

Nome Daniela Tardito 

Contatti 

Tel. 

E mail 

Tel: +390250318382 

E-mail: daniela.tardito@unimi.it 

Istituto/Dipartimento Center of Neuropharmacology, Department of Pharmacological Sciences, CEND, 

University of Milan 


Area di interesse identificata 1A. Toward exploiting the screening potential of miRNome for the 

identification of genes and pathways involved in neurodegenerative disorders 






The regulation of neuroplasticity in the response to therapeutic drugs for mood 

disorders. The role of micro RNAs 

Cariplo Foundation 

March 2010-february 2013 

Aim of this project is to investigate, by means of an integrated approach that 

merges preclinical and clinical studies with up-todate bioinformatic, 

pharmacological, genomic and genetic techniques, the role of the posttranscriptional 

regulators miRNAs in Mood Disorders aetiology and 

antidepressant response, with regard to neuroplasticity. 

Overall, the main objectives of the present project will be: 

1. to clarify the role of miRNAs in the molecular effects of different 

antidepressants on the posttranscriptional modulation of genes coding for 

modulators of neuroplasticity with an integrate approach based on preclinical 

and clinical studies. 

2. to discover new molecular targets for the development of innovative 

therapeutic strategies for the treatment of drug resistant depression 

3. to identify molecular markers associated to the antidepressant response in 

human peripheral tissues for the optimization of drug treatment in mood 

disorders. The integration of data obtained in clinical and preclinical studies will 

permit the validation of blood leucocytes as cellular models for the study of 

mental disorders 

4. to identify new susceptibility genes and pharmacogenetic markers for 

optimization of treatment in mood disorders.

Nome Maria Pennuto, PhD 

Contatti 

Phone: +39 010 71781793, Fax: +39 010 720321 

email: maria.pennuto@iit.it 

Istituto/Dipartimento Italian Institute of Technology, Department of Neuroscience and Brain 

Technology, Genoa 16163, Italy 





Mechanisms of Neurodegeneration 

Identification of signaling pathways in neurodegenerative propeinopathies 

Brain folding diseases, including Alzheimer’s disease, amyotrphic lateral sclerosis, 

and polyglutamine (polyQ) diseases, are neurodegenerative disorders 

characterized by the accumulation of toxic aggregation-prone proteins (Bertram 

and Tanzi, 2005; Ross and Poirier, 2004; Taylor et al., 2002). PolyQ diseases are a 

family of nine neurodegenerative disorders, including spinal and bulbar muscular 

atrophy (SBMA). These disorders are caused by expansion of glutamine tracts in 

nine genes, such as androgen receptor (AR). The mechanism through which the 

mutation causes neurodegeneration is not known. Despite the ubiquitous 

distribution of the disease proteins, neurons are especially vulnerable to 

expanded polyQ. Moreover, specific populations of neurons are affected in each 

polyQ disease. This suggests that the polyQ tract cannot be the sole determinant 

of disease pathogenesis. Among polyQ diseases, SBMA represents an example 

where the disease protein strikingly dictates disease features. SBMA is 

characterized by the loss of lower motor neurons and skeletal muscle atrophy. In 

SBMA, only males are fully affected. Gender-specificity is the result of the 

conversion of polyQ AR into a toxic species induced by binding to its natural 

ligand testosterone. AR is a hormone-activated transcription factor. The 

mechanism through which ligand binding converts polyQ AR into a neurotoxic 

molecule is unknown. Our long-term goal is to make a meaningful contribution 

to our understanding of the molecular mechanisms of neurodegeneration in 

brain folding diseases, so that treatment may be developed. Our objective is to 

characterize the impact of protein context, cell context, and native protein 

function on polyQ toxicity. We will test the central hypothesis that the toxicity of 

polyQ AR is influenced at the post-translational level by phosphorylation of 

specific residues in the disease protein, by tissue-specificity of disease protein 

expression, and by aberrant protein function, e.g. interaction with transcription 

co-factors and DNA. Greater insights into the molecular mechanisms underlying 

the pathogenesis of polyQ diseases is important to developing novel and 

effective therapeutic strategies for these diseases (reviewed by Pennuto and 

Fischbeck, 2009). 

TARGETING ANDROGEN RECEPTOR FOR DEVELOPMENT OF SBMA THERAPEUTICS 

Ente finanziatore Muscular Dystrophy Association (USA) 

Durata progetto 01/07/2008 to 30/06/2011 


The objectives of the MDA application are to identify post−translational 

modifications, such as phosphorylation, ubiquitylation, and SUMOylation of AR 

that reduce the toxicity of mutant protein, and to use this information to develop 

effective therapeutic interventions. We propose to accomplish these objectives 

by pursuing the following three specific Aims: 

To determine the impact of protein kinase A phosphorylation, ubiquitylation, and

SUMOylation on the toxicity of mutant AR. 

To evaluate the therapeutic efficacy of IGF−1 in vivo. 

To select an effective system to deliver IGF-1 to SBMA mice. 


POLYGLUTAMINE DISEASES: IMPACT OF PROTEIN AND CELL CONTEXT ON 

NEUROTOXICITY 

Ente finanziatore FP7−PEOPLE−2009−RG: Marie Curie International Reintegration Grant 



The objectives of this proposal are to characterize the impact of three 

components to polyglutamine toxicity: 1) protein context; 2) aberrant 

protein−protein interactions; and 3) cell context. We will test 

the central hypothesis that the toxicity of polyglutamine AR is influenced at the 

post−translational level by phosphorylation of specific residues in the disease 

protein, by aberrant interaction with the Akt effector FOXO, and by 

tissue−specificity of disease protein expression. 


INTERPLAY BETWEEN ANDROGENIC/ANABOLIC STEROID AND IGF-1 SIGNALING 

IN AMYOTROPHIC LATERAL SCLEROSIS 

Ente finanziatore Thierry Latran Foundation (France) 



The objectives of this study are to i) Determine the effect of androgens and the 

related anabolic steroid nandrolone in ALS pathogenesis; and ii) Characterize the 

molecular cross−talk between androgens and 

muscle−specific mIGF−1 in the maintenance of muscle phenotype. 


INSULIN−LIKE GROWTH FACTOR 1/AKT AND ANDROGEN SIGNALING CROSSTALK 

IN THE PATHOGENESIS OF SPINAL AND BULBAR MUSCULAR ATROPHY 

Ente finanziatore Telethon-Italy 



The objective of this application is to elucidate how phosphorylation of mutant 

AR by Akt is regulated. The central hypothesis of this application is that 

phosphorylation of mutant AR by Akt is altered by polyglutamine expansion, 

ligand binding and age and that this modification is coordinated with other 

post−translational modifications, including methylation and palmitoylation.

Nome Tanganelli Sergio 

Contatti 

tgs@unife.it 

Istituto/Dipartimento Dept. of Clinical and Experimental Medicine 

Pharmacology Section, University of Ferrara, Italy 








Mechanisms of Neurodegeneration : A. Macromolecular interactions and 

Neurodegeneration 

Neurotensin and neurotensin antagonist in an animal model of Parkinson’s 

disease: therapeutic perspectives and role of NMDA/neurotensin interaction. 

Sanofi-Aventis 

2007-2010 

The tridecapeptide neurotensin (NT) significantly enhances glutamatergic 

signaling in different brain areas and amplifies the NMDA-receptor signaling, 

probably by activating the high-affinity neurotensin receptor (NTS1), as shown by 

the NTS1receptor antagonist SR48692-induced counteraction of this effect. 

These findings suggest a reinforcing action of NT on several functions exerted by 

glutamate in the central nervous system, in particular on the glutamatemediated 

excitotoxicity. NT could be implicated in the pathogenesis of chronic 

and acute neurodegenerative disorders, such as Parkinson’s disease. To further 

investigate this hypothesis, the aims of the project, which will combine 

biochemical, functional, morphological and behavioral experiments, will be 

evaluate: 1) the putative neuroprotective effects of a pretreatment with SR48692 

(systemically administered) in an animal model of Parkinson’s disease [unilateral 

6-hydroxydopamine (6-OHDA) lesion]; 2) the functional and physio-pathological 

relevance of NMDA/NT receptor interactions in the striatum of naïve and 6- 

OHDA -lesioned rats.


B. Neurotoxic stimuli

Nome Domenico Garozzo 

Contatti 

domenico.garozzo@cnr.it 

0957338259 3473576202 

Istituto/Dipartimento CNR ICTP Catania 

Via Gaifami 18 95125 Catania 





Glycomics 

CSF Glycomics in AD and other neurodegenerative diseases 

Ente finanziatore MIUR (Italy) 

Durata progetto 3 years 

Abstract del progetto Protein glycosylation is the most common among post-translational 

modifications (PTMs), which are key to the regulation of functional 

activities of proteins. Quantitative and qualitative information about PTM 

stages of proteins is crucial in the discovery of biomarkers of disease. 

Macroheterogeneity of protein glycosylation depends on the extent of 

glycosylation site occupancy while microheterogeneity refers to 

differences in the sugar chain structures. 

Differences of protein glycosylation may be indicative of species 

synthesized in the central nervous system with respect to serum liverderived 

glycoforms, [as already demonstrated for protein Reelin, which is 

implicated in Alzheimer disease and in other neurodegenerative disorders 

(Botella-Lopez et al., 2006)]. Recent insights into neurodegenerative 

disorders caused by abnormal glycosylation of protein Seipin 

(Seipinopathy) show that modifications of Seipin glycosylation result in 

improper folding leading to accumulation of the abnormal protein in the 

endoplasmic reticulum, formation of ubiquitinated inclusion, and 

activation of UPR (Ito and Suzuki, 2009). 

Preliminary investigations based on lectin-blotting of CSF glycoproteins 

revealed quantitative modifications in Alzheimer disease and altered 

glycosylation of CSF transferrin (Taniguchi et al. 2008). We apply mass 

spectrometry based glycomics of CSF as potential tool for to detect 

possible macro-and microheterogeneity of CSF glycoproteins associated 

to Alzheimer disease. The availability of CSF depository from patients 

with AD and related disorders and established interactions with clinical 

platforms will make possible to translate the results to clinical 

proteomics. 

The objective is to find out specific protein posttranslational 

modifications related to glycosylation which in turn might be related to 

the analysed cellular phenomena associated to neurodegeneration.

Nome Maurizio Taglialatela, MD PhD – Professor of Pharmacology 

Claudio Russo, PhD – Associate Professor of Pharmacoloy 

Alfonso Di Costanzo MD – Associate Professor of Neurology 

Giovanni Scapagnini MD PhD – Associate Professor of Clinical 

Biochemistry 

Contatti Email: m.taglialatela@unimol.it Tel. +39-0874-404894/4851/4891 

Istituto/Dipartimento Dept. of Health Science, University of Molise, Via De Sanctis, 8610 

Campobasso - ITALY 



identificata 

Intracellular signalling during neurodegenerative events triggered upon APP 

overexpression. The overexpression and the aberrant processing of the amyloid 

precursor protein (APP) is a central event in the pathogenesis of Alzheimer’s Disease 

(AD). Recent data suggest that the neuronal death that occurs in different 

neurodegenerative conditions may arise from an aberrant signal related to cell cycle 

re-activation in postmitotic neurons. We are currently investigating the role of APP 

and of Aβ peptides as signalling molecules involved in MAPK and cell cycle 

activation. We propose to study a specific molecular signalling involving APP and its 

proteolytic processing by BACE1 and Presenilins, whose failure could explain at the 

same time the formation of Aβ, the induction of cell cycle and neuronal dysfunction 

observed in the development of AD. 

• Venezia V. et al., Amyloid precursor protein and presenilin involvement in cell 

signaling. Neurodegenerative Diseases. 2007, 4 (2-3): 101-111. 

• Nizzari M. et al., Amyloid precursor protein and Presenilin1 interact with the 

adaptor GRB2 and modulate ERK 1,2 signaling. The Journal of Biological Chemistry. 

2007, 282 (18): 13833-13844. 

Involvement of vascular factors as modulators of APP processing and signaling. The 

complex interaction between vascular components and tissue homeostasis is 

particularly relevant in the brain for the survival of vulnerable and irreplaceable 

postmitotic neurons. It is debated whether the presence of vascular deposits of Aβ 

peptides might represent a cause of oxidative stress and a predisposing condition 

toward the development of dementia, or rather a protective endothelial mechanism. 

In this context we are investigating the role of vascular components such as 

thrombin and apolipoproteinE as modulators of endothelial processing of APP, in Aβ 

formation and in Aβ-induced toxicity/protection. 

• Russo C. et al., Opposite roles of apolipoprotein E in normal brains and in 

Alzheimer’s disease. Proc. Natl. Acad. Sci, 1998, 95 15598-15602. 

• Russo C. et al. Presenilin-1 mutations in Alzheimer's disease. Nature. 2000; 405 

(6786):531-532. 

• Russo C. et al., The amyloid precursor protein and its network of interacting 

proteins: physiological and pathological implications. Brain Res Brain Res Rev. 2005 

48 (2): 257-264. 

Role of specific classes of ion channels in AD-related neurodegeneration. 

Intracellular potassium concentrations play a key role in cell survival. A decrease in 

cytoplasmic [K+]i, mainly caused by an increased activity of plasma membrane 

voltage-gated potassium channels, triggers cell death. Changes in K+channel activity

play a major pathogenetic role in many neurodegenerative disorders, including 

Alzheimer’s 

disease (AD). In fact, β-amyloid fragments (Aβ) alter the properties of K+ currents in 

mammalian neurons. We have described a selective modulation of a neuronal K+ 

channel (Kv3.4) upon Aβ exposure and we have shown that the inhibition of this 

channel prevents cell death triggered by the neurotoxic stimulus. We are currently 

investigating the potential neuroprotective role of Kv3.4 blockers in several models 

of AD-related neurodegeneration.. 

• Pannaccione A., et al. Nuclear factor-kappaB activation by reactive oxygen 

species mediates voltage-gated K+ current enhancement by neurotoxic betaamyloid 

peptides in nerve growth factor-differentiated PC-12 cells and 

hippocampal neurones. J Neurochem. 2005 Aug;94(3):572-86. 

• Pannaccione A, et al. Up-regulation and increased activity of KV3.4 channels and 

their accessory subunit MinK-related peptide 2 induced by amyloid peptide are 

involved in apoptotic neuronal death. Mol Pharmacol. 2007 Sep;72(3):665-73. 

Oxidative stress as a pathogenetic mechanisms and therapeutic target for 

Alzheimer Disease. Reduction of cellular expression and activity of antioxidant 

proteins leading to oxidative stress are fundamental causes for brain aging and 

neurodegenerative diseases. Deregulation of the antioxidant protein Heme 

oxygenase-1 (HO-1) has been associated with the pathogenesis of Alzheimer's 

disease, multiple sclerosis and brain aging. Moreover, a number of experimental and 

epidemiological studies have recently supported the beneficial effects of some 

commonly used natural products such as curcumin, possibly by increasing the 

activity of the HO-1 protein. Curcumin has been reported to decrease oxidative 

damage and amyloid deposition in a transgenic mouse model of Alzheimer's disease, 

and to reverse Aβ-induced cognitive deficits and neuropathology in rats. We are 

currently investigating the potential neuroprotective role and the underlying 

mechanisms, of HO-1-inducing natural compounds in various in vitro and in vivo 

models of AD. 

• Scapagnini G. et al., Caffeic acid phenethyl ester and curcumin: a novel class of 

hemeoxygenase-1 inducers. Mol Pharmacol. 2002, 61(3):554-61. 

• Scapagnini G. et al., Ethyl ferulate, a lipophilic polyphenol, induces HO-1 and 

protects rat neurons against oxidative stress. Antioxid Redox Signal. 2004, 6(5):811- 

8. 

• Scapagnini G. et al., Curcumin activates defensive genes and protects neurons 

against oxidative stress. Antioxid Redox Signal. 2006, 8(3-4):395-403. 

Physical exercise and cognitive decline. Epidemiological studies have shown that 

physical exercise can delay the occurrence and the progression of dementia, but also 

to improve physical, cognitive and psychological performances, with a positive 

impact on the quality of life. The mechanisms involved and the identification of 

potential biomarkers predictive of a positive response to physical activity are areas 

of intense investigation. In our group, we are currently investigating the potential 

preventive role of physical exercise programs on cognitive decline in subjects at risk 

to develop dementia. Our focus will be on subjects presenting with a subjective 

memory disturbance or affected by mild cognitive impairment (MCI). 

• Tedeschi G., et al. Brain atrophy and lesion load in a large population of patients 

with multiple sclerosis. Neurology. 2005 Jul 26;65(2):280-5. 

• Tedeschi G., et al. Correlation between fatigue and brain atrophy and lesion load 

in multiple sclerosis patients independent of disability. J Neurol Sci. 2007 Dec 

15;263(1-2):15-9.


Titolo 

progetto 

Ente 

finanziatore 

Durata 

progetto 

Abstract del 

progetto 

Titolo 

progetto 

Ente 


Durata 

progetto 

Abstract del 

progetto 

Regulation of K + channels by APP-interacting proteins: functional characterization and 

implications for AD-related neurodegeneration 

Regione Campania (2000-2003) 

3 years 

Recent experimental evidence suggest that K + channels play a major pathogenetic role in 

Alzheimer disease. In the CNS, phosphorylation of voltage-gated K+ channels by tyrosine 

kinases (PTK) is a fundamental process regulating their functional activity; its inhibition 

exerts a neuroprotective action in several experimental models of neuronal damage. On 

the other hand, the expression of several subclasses of K + channels can reduce cellular 

tyrosine kinase activity, thus suggesting the existence of a reciprocal interaction between 

PTKs/PTPases and K + channels. Given that the cytosolic domain of β-APP influences, 

through its adaptor proteins Dab (disabled), X11 and Fe65, the tyrosine kinase activity of 

Abl, in the present proposal we will: 

A. Evaluate the potential modulation of several classes of K + channels (Kv1.2, Kv1.3, Kv1.5, 

Kv2.1, ERG, KCNQ2+3) by adaptor proteins able to bind to β-APP (X11, Fe65, Dab). 

B. Verify the participation of tyrosine kinase activity (and in particular of Abl), in this 

modulation. 

C. Investigate the potential involvement of these channels in the cellular damage induced 

by the hyperexpression of β-APP adaptor proteins. 

D. Study the participation of K + channels and their regulation by oxidative stress in the 

neuronal death triggered by various cellular models of AD-related neurodegeneration. 

European Community contract N° LSHM-CT-2003-503330/Apopis : Abnormal proteins in 

the pathogenesis of neurodegenerative disorders 

Study on tyrosine phosphorylation of APP and its interaction with intracellular adaptors: 

role in cell signaling and in the generation of amyloidogenic fragments. 

European Community 

3 years (2004-2007) 

Within the WP 1 Generation and Turnover, our proposal is aimed to the definition of the 

role that posttrasductional modifications of APP, in particular its phosphorylation, may 

exert on its pathophysiologycal function. We will investigate both the role of 

phosphorylated APP/CTFs in cell signaling, and/or in the generation of amyloidogenic 

fragments and plaques through the interaction with intracellular adaptors, as well as the 

effect that site-specific tyr-phosphorylation would have on the direct amyloidogenic 

pathway of APP. These studies, which are focused in a completely new aspect of APP 

activity, would lead to the identification of early markers of the neurodegenerative process 

which occurs in AD, and to the definition of a potentially innovative therapeutical approach 

based on the findings obtained. Our proposal is aimed at: 

Aim 1) To study directly in human brain the tyr-phosphorylation of APP and its CTFs and 

their possible coupling with intracellular adaptors such as ShcA, and to correlate these data 

with the presence of neuropathologycal hallmarks of the disease. The comparative analysis 

in non-AD control, AD and/or Down’s syndrome subjects at different ages (from fetal to 

adult life) will allow a deeper characterization of the molecular determinants during the

progression of the disease. 

Aim 2) To study in transgenic mice carrying APP and PS1 mutations the tyrphosphorylation 

of APP and CTFs, their interaction with ShcA-Grb2 adaptors and the 

influence that such interaction could have on plaque formation, astroglial response, 

neuronal death. The use of a Tg model will allow at a deeper comprehension of the 

possible correlation between APP phosphorylation, its cleavage, signaling activity and 

progressive presence of the typical hallmarks of the disease. Moreover, in this model is 

possible also to determine the effect that APP and PS1 mutations, which are linked with a 

human familial phenotype, would cause in the above mentioned parameters, and, in a 

second step, would allow the application of targeted therapeutical pharmacological 

approaches following also the indications raised from the in vitro studies (aim 3). 

Aim 3) To study in neuronal and/or glial cell cultures the molecular mechanisms which 

regulate APP phosphorylation, influence on cleavage by secretases, coupling with 

intracellular adaptors, and signaling activity. Using APP mutants, ShcA mutants, and APP 

KO cells we will define the influence that phosphorylation and interaction with ShcA would 

cause in signaling activity, cell proliferation and death, and in the generation of 

amyloidogenic fragments. Moreover, the study of the effect that proliferative and 

apoptotic stimuli may cause on those parameters and/or with the study of the kinases 

involved in the activation of APP cleavage and signaling, this would also lead to the 

identification of pharmacological target for a therapeutical intervention.

Nome MARIO MARCHI 

Contatti marchi@pharmatox.unige.it 

Istituto/Dipartimento Department of Experimental Medicine (Di.Me.S.) 

Section of Pharmacology and Toxicology 

Viale Cembrano, 4 16147 Genova ITALY +39 010 3532657 e.mail: 


Effect of Age and neurodegenerative disorders on central neurotransmission: focus on neurotransmitter 

release from neurons and glia 

The effects of age and/or neurodegenerative disorders may differentially influence diverse cell types in 

the brain and the vulnerability of neurons may depend also on the transmitter system or the location in 

neural circuits. Therefore, it is of utmost importance to identify age-induced neurochemical changes in the 

brain in order to develop pharmacological strategies that can prevent, delay or counteract the cognitive 

decline. 

Cells may be differentially sensitive according to the function of signaling pathways that mediate cell 

specific processes involved in cognition (e.g. synaptic plasticity). Systemic influences of inflammation can 

also alter the local milieu to influence different cell populations. Our current researches are focused to 

assess the current status of synaptic function/dysfunction in aging and age-related neurodegenerative 

diseases in several regions of the brain in mammals, particularly in rodents .It is now well accepted that 

astrocytes integrate and process synaptic information and control synaptic transmission and plasticity 

being active partners in synaptic function, astrocytes are cellular elements involved in the processing, 

transfer and storage of information by the nervous system. Therefore our studies are aimed at 

investigating “ex-vivo” changes of neuronal and glial functions due to “in-vivo” aging or in the animal 

models of neurodegenerative disorders (i.eAlzheimer;) The pivotal concept of this approach assumes that 

“in-vivo” aging or the neurodegenerative disorders are able to modify the plasticity of chemical synapses 

which in turn could be retained, and then analyzed, in the “ex-vivo” tissue preparations. This “ex-vivo” 

memory has been already described to occur and studied in our laboratory to unmask neurotransmission 

modifications caused by “in-vivo” exposure to different stimuli.( i.e. chronic treatment with drugs, 

enriched environment, etc.). Our research is therefore focused to investigate whether and to what extent 

aging and age-related neurodegenerative diseases (i.e. Alzheimer) could modify some fundamental 

parameters, such as neurotransmitter release and uptake, receptor expression and functioning in plasma 

membranes, moreover we will investigate the effect of beta amyloid on these parameters by utilizing two 

“ex-vivo” functional preparations representative of neurons and astrocytes , i.e. the isolated nerve 

terminals (synaptosomes) and the glial subcellular particles (gliosomes). 




Plasticità dei recettori presinaptici nicotinici e glutammatergici che 

modulano la liberazione di neurotrasmettitori: studio delle variazioni 

delle loro risposte funzionali dopo trattamenti cronici o in seguito ad 

interazioni con altri sistemi recettoriali o altre condizioni sperimentali 

Role: Principal Investigator 

Ente finanziatore MIUR-prin 



24 MESI 


Studies on the effect of the chronic treatment with nicotine on some

neurochemical presynaptic mechanisms and on the function of 

presynaptic receptors in the CNS. 

Role: Principal Investigator 

Ente finanziatore COMPAGNIA DI SAN PAOLO 

Durata progetto 36 MESI 

Titolo progetto Les plant aromatique, entre environment et activitè productive 

Role . Principal Investigator 

Ente finanziatore ALCOTRA “AROMA” 

Durata progetto 24 MESI

Nome Dr. Francesca Ruberti 

Contatti 

f.ruberti@inmm.cnr.it; INMM- CNR Istituto di Neurobiologia e Medicina 

Molecolare Via del Fosso di Fiorano, 64 00143 ROMA ITALY Phone: + 39 

06 501 703 236 Fax: + 39 06 501 703 313 

Istituto/Dipartimento Institute of Neurobiology and Molecular Medicine CNR 


NEUROBIOLOGY OF ALZHEIMER’S DISEASE 

1) Basic research : developing competitive in vitro and in vivo models to study Alzheimer’s Disease 

a) Role of microRNAs in Alzheimer’s Disease onset and progression (C. Barbato, C Cogoni, F. Ruberti) 

Primary neuronal cell culture and Alzheimer’s Disease (AD) mouse models, including AD11 (and transgenic 

mice overexpressing pathogenic variants of human APP gene, will be instrumental to investigate the 

function of selected miRNAs (by their overexpression and/or inhibition) on disease pathogenesis. Studies 

will focus on recently identified microRNAs modulating, in neuronal hippocampus cultures (Vilardo et al. 

2010), APP production and on miRNAs dysregulated in human AD brain (Barbato et al 2009). 

Multidisciplinary approaches will be used to investigate the effect of microRNAs on molecular mechanisms 

underlying amyloidogenesis, neuronal degeneration, memory loss, synaptic plasticity impairment. 

Vilardo E, Barbato C, Ciotti MT, Cogoni C, Ruberti F. MicroRNAs regulate Alzheimer’s Amyloid Precursor 

Protein expression in hippocampal neurons. (2010) submitted. 

Barbato C, Ruberti F, Cogoni C. Searching for MIND: microRNAs in neurodegenerative diseases. J Biomed 

Biotechnol. 2009:871313. 

Barbato C, Ruberti F, Pieri M, Vilardo E, Costanzo M, Ciotti MT, Zona C, Cogoni C. MicroRNA-92 modulates 

K(+) Cl(-) co-transporter KCC2 expression in cerebellar granule neurons. J Neurochem. 2009 Dec 26. 

b) Mechanisms underlying the effect of N-terminal 26-230 tau fragment on synaptic dysfunction and 

neurodegeneration (N. Canu, in collaboration with V. Cestari, F. Tirone, E Mattei). The aim is is to decode 

the signal transduction pathway that links N-terminal 26-230 tau fragment (N-tau) expression to NMDAR 

activity (Canu et al 1998; Amadoro et al 2006) and their upstream and downstream effectors inducing 

synaptotoxicity and plasticity failure. Electrophysiological recording of NMDAR and AMPAR biochemical 

characterization of NMDAR complex, morphological, qualitative and quantitative analysis of synapse 

number, dendritic spines and behavioral analysis of conditional N-tau transgenic mice, will be performed 

to give an accurate and global view of the molecular events involved in N-tau modulation of neuronal 

plasticity. 

Canu N, Dus L, Barbato C, Ciotti MT, Brancolini C, Rinaldi AM, Novak M, Cattaneo A, Bradbury A, Calissano 

P. Tau cleavage and dephosphorylation in cerebellar granule neurons undergoing apoptosis. J Neurosci. 

1998 Sep 15;18(18):7061-74. 

Amadoro G, Serafino AL, Barbato C, Ciotti MT, Sacco A, Calissano P, Canu N. (2004) Role of N-terminal tau 

domain integrity on the survival of cerebellar granule neurons. Cell Death Differ. 2004 Feb;11(2):217-30. 

Amadoro G, Ciotti MT, Costanzi M, Cestari V, Calissano P, Canu N. NMDA receptor mediates tau-induced 

neurotoxicity by calpain and ERK/MAPK activation.Proc Natl Acad Sci U S A. 2006 103(8):2892-7 

c) Nerve Growth Factor deprivation pathways leading to Alzheimer’s Disease phenotype (C. Matrone, P. 

Calissano). An NGF-dependent hippocampal neuronal model in which the interruption in NGF supply 

quickly triggers amyloidogenesis and induces tau protein dysfunction and neuronal degeneration, 

mimicking essential features of in vivo AD pathology, has been recently charachterised (Matrone et al. 

2008; Matrone et al 2009; Amadoro et al 2009). The main goal is to to test whether the proteins whose 

expression or phosphoylation is affected by NGF deprivation may represent early molecular signature and 

potential targets of AD. To achieve these results, studies are in progress, in collaboration with prof. Moses 

Chao at NYU, to characterize the phosphotyrosine-associated signalling events due to NGF deficit in 

primary cultures from wild type and APPk/o mice by a systematic MS/SILAC.

Matrone C, Ciotti MT, Mercanti D, Marolda R, Calissano P. NGF and BDNF signaling control amyloidogenic 

route and Abeta production in hippocampal neurons.Proc Natl Acad Sci U S A. 2008 105:13139-44. 

Matrone C, Marolda R, Ciafrè S, Ciotti MT, Mercanti D, Calissano P. Tyrosine kinase nerve growth factor 

receptor switches from prosurvival to proapoptotic activity via Abeta-mediated phosphorylation. Proc Natl 

Acad Sci U S A. 2009 106: 11358-63. 

Amadoro G, Corsetti V, Ciotti MT, Florenzano F, Capsoni S, Amato G, Calissano P. Endogenous Abeta causes 

cell death via early tau hyperphosphorylation. Neurobiol Aging. 2009 Jul 21. 

d) Role of NH2-26-44 tau peptide in Alzheimer’s Disease (G. Amadoro P Calissano). A synthesized NH2-26- 

44 peptide of tau, impairs the mitochondrial oxidative phosphorylation and reduces the intracellular ATP 

bioavailability, probably acting on adenine nucleotide translocator (ANT)- mediated ADP/ATP exchange 

(Atlante et al., 2008). Studies in AD human tissues are focused to evaluate if tau peptide i) is correlated 

with the extent of neurofibrillary degeneration and amyloid neuropathology, especially Abeta pre-fibrillar 

species; (ii) is linked to the synaptic changes and to the mitochondrial functional alterations. The physical 

interaction between ANT and NH2-derived tau fragment, in pathologically relevant conditions, will be 

investigated. 

Atlante A, Amadoro G, Bobba A, de Bari L, Corsetti V, Pappalardo G, Marra E, Calissano P, Passarella S. A 

peptide containing residues 26-44 of tau protein impairs mitochondrial oxidative phosphorylation acting at 

the level of the adenine nucleotide translocator. Biochim Biophys Acta. 2008 1777, 1289-300 

e) Mitochondrial damage and genetic risk/susceptibility factors in Alzheimer’s Disease (M. Rinaldi, S. 

Iurescia, D. Fioretti). The aim of the research project is i) to identify and characterize early biomarkers and 

mechanisms of mitochondrial damage promoted by ROS/RNS; ii) to investigate the protective role of 

natural forms and synthetic analogues of vitamin E that can be tested in intervention studies in AD 

patients. Studies are in progress to evaluate post-transductional modifications (PTMs) in mitochondrial 

proteins, caused by ROS/RNS in in vitro model of senescence and human neuroblastoma as well as in 

peripheral cells from subjects with Mild Cognitive Impairment (MCI) and AD. Strong genetic 

risk/susceptibility factors for developing Alzheimer’s disease, such as ApoE4 that impacts on Abeta 

production, Abeta clearance, Abeta fibrillation and tangle formation as well as on mitochondrial functions 

leading to neuronal toxicity and synaptic damage, are also investigated. 

Iurescia S, Fioretti D, Mangialasche F, Rinaldi M.The Pathological Cross Talk Between Apolipoprotein E and 

Amyloid-beta Peptide in Alzheimer's Disease:Emerging Gene-Based Therapeutic Approaches. J Alzheimers 

Dis. 2010 In press 

2) Basic research : New treatment strategies based on Neurotrophins 

f) Ocular NGF administration as a novel non invasive approach to protect brain-NGF target neurons that 

degenerate in Alzheimer’s disease (L. Aloe, P. Tirassa). Nerve growth factor and (NGF) is a soluble protein 

that plays a protective role on brain neurons that degenerate in age-related brain disorders, including 

Alzheimer’s disease (AD). We have shown that conjunctively applied NGF can protect injured brain 

neurons and damaged retinal ganglion neurons suggesting that topical eye NGF application might be a 

novel alternative for delivering NGF into the brain and for protecting brain neurons and reducing memory 

deficits occurring during early events of AD and glaucoma. This hypothesis is currently actively under 

investigation in collaboration with other research groups. 

Lambiase A, Pagani L, Di Fausto V, Sposato V, Coassin M, Bonini S, Aloe L. Nerve growth factor eye drop 

administrated on the ocular surface of rodents affects the nucleus basalis and septum: biochemical and 

structural evidence. Brain Res. 2007;1127:45-51. 

Di Fausto V, Fiore M, Tirassa P, Lambiase A, Aloe L. Eye drop NGF administration promotes the recovery of 

chemically injured cholinergic neurons of adult mouse forebrain. Eye drop NGF administration promotes 

the recovery of chemically injured cholinergic neurons of adult mouse forebrain. Eur J Neurosci. 

2007;26:2473-80. Lambiase A, Aloe L, Centofanti M, Parisi V, Mantelli F, Colafrancesco V, 

Manni GL, Bucci MG, Bonini S, Levi-Montalcini R. Experimental and clinical evidence of neuroprotection by

nerve growth factor eye drops: Implications for glaucoma. Proc Natl Acad Sci U S A, 2009; 109: 13469- 

13474,. 

g) Electro-acupuncture and polyphenols as novel approaches to reduce neurodegeneration ( L Manni, M 

Fiore). Preliminary data indicate that diabetes induced in adult rats by injection with streptozotocin causes 

a decrease of brain NGF and a concomitant increase of tau phosphorylation, that were all counteracted by 

low-frequency electro-acupuncture (EA). Using molecular and behavioural approaches the research 

project will be aimed at: i) Investigate the possible link between diabetes, brain NGF presence/activity and 

the development of AD-associated pathological features; ii) Study the effect of EA on the above mentioned 

diabetes-associated brain dysfunction. Alcoholism is related to neurodegeneration and diabetes. The use 

of polyphenols extracted by olive oil, olive leaves and olive seeds or by grapes (Fiore et al 2009) to prevent 

or limit mouse brain neurodegeneration due to aging or ethanol consumption will be evaluated. 

Manni L, Aloe L, Fiore M. Changes in cognition induced by social isolation in the mouse are restored by 

electro-acupuncture. Physiol Behav. 2009;98(5):537-42 

Fiore M, Laviola G, Aloe L, di Fausto V, Mancinelli R, Ceccanti M. Early exposure to ethanol but not red wine 

at the same alcohol concentration induces behavioral and brain neurotrophin alterations in young and 

adult mice. Neurotoxicology. 2009;30(1):59-71. 

h) Human neural stem cells lentivirally transduced with human BDNF (C. Cenciarelli, P. Casalbore). A 

strong body of evidences show that BDNF result critically involved in Alzheimer’s and Huntington’s 

diseases. Recently, it has been shown that BDNF knockdown within NSC abolishes the cognitive benefits of 

NSC delivery. The aim is to use propagating human neural stem cells (hNSC) lentivirally transduced with 

human BDNF as cellular therapy for replacing degenerating neurons in disease, trauma and toxic insults. 

Cenciarelli C, Budoni M, Mercanti D, Fernandez E, Pallini R, Aloe L, Cimino V, Maira G, Casalbore P. In vitro 

analysis of mouse neural stem cells genetically modified to stably express human NGF by a novel multigenic 

viral expression system. Neurol Res. 2006, 28:505-12. 

Casalbore P, Barone I, Felsani A, D’Agnano I, Michetti F, Maira G and Cenciarelli C. Neural stem cells 

modified to express BDNF antagonize trimethyltin-induced neurotoxicity through PI3K/Akt and MAP kinase 

pathways . Accepted on J. Cell. Physiol. 2010. 

OTHER NEURODEGENERATIVE DISEASES 

Basic research, biobanking (DNA), sharing database, exchanging and pooling data 

i) Biobank compund by over 2000 DNAs from HD and ataxic patients and their families. Molecular basis 

of episodic ataxia (L. Veneziano). Research activity has been mostly devoted to CACNA1A gene, coding for 

the Alpha1A subunit of P/Q type Calcium channel expressed in brain, from its physical map to the 

refinement of the gene structure to the screening for mutations in Episodic Ataxia (EA) to the association 

genotype/phenotype (Veneziano et al 2009; Mantuano et al 2010). This research allowed the construction 

of a DNA biobank from about 100 patients affected by EAs. The project is focused on the use Comparative 

Genomic Hybridization (CGH) arrays, which can be used for a screening of CACNA1A gene rearrangement 

in a sample of 40 EA patients in which CACNA1A gene point mutations were excluded. The fulfilment of 

the above aim will improve the rate of molecularly diagnosed EA2 patients to be treated with known 

therapy or to be included in new therapeutic trials. 

Veneziano L, Guida S, Mantuano E, Bernard P, Tarantino P, Boccone L, Hisama FM, Carrera P, Jodice C, 

Frontali M. Newly characterised 5' and 3' regions of CACNA1A gene harbour mutations associated with 

Familial Hemiplegic Migraine and Episodic Ataxia. J Neurol Sci. 2009, 276(1-2):31-7 

Mantuano E, Romano S, Veneziano L, Gellera C, Castellotti B, Caimi S, Testa D, Estienne M, Zorzi G, Bugiani 

M, Rajabally YA, Barcina MJ, Servidei S, Panico A, Frontali M, Mariotti C.Identification of novel and 

recurrent CACNA1A gene mutations in fifteen patients with episodic ataxia type 2. J Neurol Sci. 2010 Feb 1. 

l) Role of the Thyroid Transcription Factor 1 (TTF-1) in neurodegenerative diseases (D. Civitareale). The 

Thyroid Transcription Factor 1 (TTF-1) expression persists in some post-mitotic basal forebrain neurons, 

and mutations of TiTF-1 have been associated with the benign hereditary chorea. Our transcriptome 

analysis confirms that TTF-1 is involved in neuron morphogenesis, differentiation and GnRH secretion. We 

predict that TTF-1 could be a modifier gene involved in the pathogenesis of the neurodegenerative 

diseases. Therefore, we propose to study TTF-1 down-regulation via RNAi and microarray analysis in

ST14A, select a mouse strain able to develop the neurodegenerative disease in absence of Titf-1, and 

characterize the genotype/phenotype association of TiTF-1 in a cohort of patient with neurodegenerative 

diseases. 

Provenzano C, Veneziano L, Appleton R, Frontali M, Civitareale D. Functional characterization of a novel 

mutation in TITF-1 in a patient with benign hereditary chorea. J Neurol Sci. 2008 264(1-2):56-62. 

Provenzano C, Pascucci B, Lupari E and Civitareale D. Large scale analysis of transcription factor TTF- 

1/NKX2.1 target genes in GnRH secreting cell line GT1-7. Molecular and Cellular Endocrinology. In press. 


Basic research : developing competitive in vitro and in vivo models to 

study Alzheimer’s Disease; new treatment strategies based on 

Neurotrophins. Biobanking 

(DNA), sharing database, exchanging and pooling data 

Finanziamenti ricevuti Role of microRNAs in Alzheimer’s Disease onset and progression (C. Barbato, C 

Cogoni, F. Ruberti,). 

Ongoing Research Support -3886 

SD/sd 2008.2404 Programma Neuroscienze “Compagnia di San Paolo” 9/01/09-9/01/11: 

‘MIND: MicroRNA In Neurodegenerative Diseases’ (to C. Barbato) 

-REGIONE LAZIO grant 2009-2011: ‘Studio delle basi molecolari della neurodegenerazione 

nella malattia di Alzheimer’- (to C.Cogoni). 

Completed Research Support - 

DG.RSTL.059.012 –CNR grant-01/2008-12/2009. “Study of APP regulation by microRNAs 

and implication in Alzheimer’s Disease” (to F. Ruberti) 

Finanziamenti ricevuti Mechanisms underlying the effect of N-terminal 26-230 tau fragment on synaptic 

dysfunction and neurodegeration (N. Canu). 

Ongoing Research Support - 

Dipartimento di Neuroscience, University of Tor Vergata 

Completed Research Support - 

MIUR-PRIN 2006-2008: “Molecular and behavioural analysis of tau neurotoxic effect in 

cellular and animal models” 

-Ministero della Sanità- Ex Articolo 56 Neurodegenerativo 533F/B/1 2005-2009 

“Identificazione di marcatori biologici precoci per la demenza di Alzheimer: genomica e 

proteomica nella neurodegenerazione” 

Finanziamenti ricevuti Nerve Growth Factor deprivation pathways leading to Alzheimer’s Disease 

phenotype (C. Matrone, P. Calissano). 


MIUR-FIRB Italy / 2009-2011 

“Italian Human Proteome Net” - 

MIUR-PRIN 2009/2011 

“Ruolo di Zn e Cu nella cascata amiloidogenica provocata da deprivazione di NGF” 

Finanziamenti ricevuti Role of NH2-26-44 tau peptide in Alzheimer Disease (G. Amadoro P. Calissano) 


MIUR-FIRB 24-09-08 al 24-09-11: “Italian Human Proteome Net” - 

MIUR 18-07-2007 to 18-07-2010: “Neurotrophic and neuroprotective drugs suitable for 

therapeutic applications in neuroscience: AD,HD,SLA” 

Finanziamenti ricevuti Mitochondrial damage and genetic risk/susceptibility factors in Alzheimer’s

Disease (M. Rinaldi, S. Iurescia, D. Fioretti). 

Ongoing Research Support MIUR- 

PRIN 2007-2009: “Oxidative and nitrosative damage of mitochondrial proteins in models 

of cell senescence and in subjects with Alzheimer's disease: a pre-clinical study on natural 

forms and new synthetic analogues of vitamin E. 

Finanziamenti ricevuti, Ocular NGF administration as a novel non invasive approach to protect brain- 

NGF target neurons that degenerate in Alzheimer’s disease (P Tirassa, L Aloe) 


Progetto PRIM- 2007-2010: “Sviluppo del NGF come farmaco in collirio: Studi di 

farmacocinetica e di efficacia terapeutica”. (to L. Aloe) - 

Progetto Strategico Q77, ISS- 2007-2010: “Indicazioni di indici neurobiologici di 

vulnerabilità a rischio di sviluppo di sindromi depressive” (to L. Aloe). - 

Fondazione G.B. Bietti 2007-2009: “Potenziale ruolo terapeutico del NGF nel glaucoma: 

Studi in un modello animale e nell’uomo” (to L. Aloe) - 

CNR funding (PORRSTL03) 2006-2008 (to P. Tirassa) 

Finanziamenti ricevuti, Electro-acupuncture and polyphenols as novel approaches to reduce 

neurodegeneration (L Manni, M Fiore) 

Ongoing Research Support Centro 

Alcologico Regione Lazio (to M. Fiore) 

Finanziamenti ricevuti, Human neural stem cells (hNSC) lentivirally transduced with human BDNF (C. 

Cenciarelli, P. Casalbore). 

Ongoing Research Support Funds 

from ATENA Onlus (2009-2010): “Transplantation of BDNF secreting-NSC in the 

hippocampus of adults rats treated with the neurotoxicant thrimetyltin.” 

Completed Research Support Grants 

from The Nando Peretti Foundation and ATENA Onlus (2006-2008) “Repair of 

lesions of Central Nervous System through cell replacement therapy by using of NSC 

induced towards to neuronal phenotypes.” 

Finanziamenti ricevuti, Biobank compund by over 2000 DNAs from HD and ataxic patients and their 

families and Molecular basis of episodic ataxia (L. Veneziano). 

Ongoing Research Support National 

Ataxia Foundation 2009: “Looking for gross rearrangements of CACNA1A gene in Episodic 

Ataxia type 2 (EA2) patients by Comparative Genomic Hybridization arrays.

Nome MARIAROSARIA MILOSO 

Contatti mariarosaria.miloso@unimib.it tel 0264488123 

Istituto/Dipartimento 


Dipartimento di Neuroscienze e Tecnologie Biomediche 

Area di interesse identificata EVALUATION OF MESENCHYMAL STEM CELLS (MSCs) EFFECT IN 

ALZHEIMER’S DISEASE RAT MODELS: in vitro and in vivo STUDIES 

Alzheimer’s disease (AD) is the most common cause of progressive 

decline of cognitive function in aged humans. At present, there are no 

effective therapies to treat this pathology. Recently it has been 

hypothesized an innovative therapeutic approach based on the use of 

stem cells. Different sources of stem cells are being explored for potential 

use in repairing neurodegenerative disorders. Among them, 

Mesenchymal Stem Cells (MSCs) represent a good cellular source for 

transplantation therapy. MSCs can be easily isolated and expanded, 

autologously used, present a low risk of tumorigenicity and are 

ipoimmunogenic. 

Aim of our project is to study the possible effect of human and rat MSCs 

in an in vitro and in vivo model of Alzheimer Disease (AD). 

In vitro model of AD is represented by rat cortical and hippocampal 

neuron primary cultures exposed to 2,5 µM Aβ (1-42) fibrils. To evaluate 

the capacity of MSCs to improve neuronal viability and AD 

neuropathological changes, we will perform: a) direct and indirect co- 

cultures of MSCs with Aβ-treated neurons; b) culture of Aβ-treated 

neurons in the presence of conditioned medium from MSCs. Neuronal 

survival will be evaluated by MTT assay, neuronal morphology and 

functionality will be assessed by morphological and immunofluorescence 

studies and electron microscopy. 

In vivo model of AD is represented by a transgenic rat model that develop 

spontaneously the AD pathology. The MSCs are prelabeled with Qdot 

nanocristal and than inoculated into the rat tail vein. After the MSCs 

injection cognitive functions will be assessed by the Morris water maze 

test (Morris 1984). After the behavioral test, the rats will be sacrificed 

and the brain will be obtained and further fixed. Sections of cortex and 

hippocampus will be cut in a freezing microtome and AD






neuropathological changes will be evaluate by morphological and 

immunofluorescence studies and electron microscopy. MSC localization 

and differentiation will be evaluated by immunofluorescence 

experiments.

Nome Patrizia Hrelia, Full Professor in Toxicology, Faculty of Pharmacy 

Contatti 

Via Irnerio 48, 40126 Bologna 

Tel.: +39 051 2091799 

e-mail: patrizia.hrelia@unibo.it 

Istituto/Dipartimento Department of Pharmacology , Alma Mater Studiorum - University of 

Bologna 

Nome Silvana Hrelia, Associate Professor in Biochemistry, Faculty of 

Pharmacy 

Contatti 

Via Irnerio 48, 40126 Bologna 

Tel.: +39 051 2091233 

e-mail: silvana.hrelia@unibo.it 

Istituto/Dipartimento Department of Biochemistry, Alma Mater Studiorum - University of 

Bologna 


Our project is aimed at the identification of cellular and molecular targets affected by 

phytochemicals, to delineate pharmacological and nutritional strategies for the reduction or the 

prevention of neurodegeneration and neuroinflammation associated to ageing and 

neurodegenerative diseases. The project is innovative since it represents a joint action with the 

attempt: 

1) to adequately understand the cellular and molecular mechanisms commanding cell death in 

neurodegeneration and behind neuroprotective effects of phytochemicals; 

2) to identify phytochemicals having diverse pharmacological activities, such as neuroprotection and 

antiinflammation; 

3) to confirm if these phytochemicals have a specific pharmacological effectiveness in the 

prevention/counteraction of one of the most important aging-related neurodegeneration and 

neuroinflammation hallmarks, oxidative stress, as a primary prevention mechanism; 

4) to understand if these phytochemicals can act as biomodulators in neuronal cells through 

modulation of survival pathways, suggesting a potential therapeutic use in regenerating injured 

neuronal cells; 

5) to establish the therapeutic window that allows an effectively impact of phytochemicals on 

specific targets; 

In longer terms, our studies will indicate whether pharmacological and nutritional strategies with 

phytochemicals should be effective and safe in counteracting cellular and molecular damage 

associated to human neurodegeneration and neuroinflammation.

Area di interesse identificata New treatment strategies (Neurodegeneration and 

Neuroprotection) 



Drug Research and Development (CHEM-PROFARMA-NET). 

Synthesis, biological and pharmacological characterization of 

organic molecules, bio-oligomers and natural compounds endowed 

with antidegenerative (neuro or cardiovascular) antiviral, and antiinfective 

activity. 

Ente finanziatore MIUR FIRB – Piattaforme/Reti (total funds euro9.768.680, 

euro1.274.097 assigned to P.Hrelia as node coordinator) 

Durata progetto 2007-2011 


The vision of the project is that of a strong collaborative efforts 

aimed at the identification of new cellular and molecular targets 

affected by synthetic or natural compounds to delineate 

pharmacological and nutritional strategies for the prevention of 

human diseases such as neurodegeneration, infectious disease 

cancer, immune and metabolic disorders. The activities proposed 

will be completely performed in the Network Operative Units. This 

project is aimed at identifying new drugs that may be useful in 

preventing and/or alleviating the consequences of specific diseases, 

but it will focus on new therapeutic approaches for such disorders. 

Titolo progetto New Bioactive Molecules from Natural Sources in Neuroprotection: 

Opportunities In Neurodegenerative Diseases 

Ente finanziatore MIUR COFIN 2007 (total funds euro 63.750 assigned to G. Cantelli 

Forti as Scientific Coordinator, and S. Hrelia as Responsible of 

Research Unit) 



The pathogenesis of ageing related neurodegenerative diseases, 

such as Alzheimer's (AD) or Parkinson's (PD), is multifactorial with 

the existence of a "domino" cascade of neurotoxic events, which 

can be initiated at any point in the cascade. Many lines of evidence 

suggest that oxidative stress, resulting in ROS/RNS generation, 

mitochondrial dysfunction and activation of death/survival 

pathways play a pivotal role in the age-associated neuronal loss in 

neurodegenerative diseases. Thus, promising future treatment of 

neurodegenerative diseases depends on availability of effective 

brain permeable, antioxidant neuroprotective drugs that would 

prevent the progression of neurodegeneration. An innovative 

approach to preventive/therapeutic intervention in oxidative stress 

based neurodegeneration may be the manipulation of endogenous

cellular defence mechanisms by chemical inducers, such as 

isothiocyanates from natural sources, among which sulforaphane 

(SF). Molecular and cellular mechanisms behind SF neuroprotection 

will be initially studied in vitro, through an experimental approach 

which let us to evaluate neuroprotective and/or neurorescue effects 

by SF. Neuronal damage parameters at the cellular level will 

comprise the induction of neuronal death in terms of 

apoptosis/necrosis and the activation of cellular death/survival 

biochemical pathways. The scientifically important expectation of 

this project, by providing data on the cellular and molecular 

mechanisms commanding cell death in neurodegeneration and 

behind neuroprotection by SF, is to provide a clear rationale for 

delineating innovative pharmacological strategies to counteract and 

rescue neurodegeneration.

The research group 

Fondazione IRCCS Istituto Neurologico “Carlo Besta” 

Division of Neuropathology – Neurology 5 

Director: Fabrizio Tagliavini 

The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the diagnosis and treatment of 

patients with degenerative dementias (over 2000 out-patients and 150 in-patients per year) and (ii) a 

Laboratory Unit dedicated to the analysis of genes and biomarkers associated with degenerative 

dementias, post-mortem characterization of the disease process and disease-specific protein, and 

experimental studies on disease pathogenesis and the development of therapeutic strategies. The course 

of action of this activity can be expressed as a “Bed to Bench to Bed” cycle, with the ultimate goal to 

identify disease-modifying drugs in preclinical settings and apply them to patients. In this regard, the 

Clinical Unit is currently coordinating a multicentre phase II clinical trial supported by AIFA, to test the 

effectiveness of an anti-amyloidogenic molecule identified in experimental models. This comprehensive 

approach has also the great advantage to collect biological samples (plasma, DNA, CSF and brain tissue) 

from fully characterized patients. In this regard the Laboratory Unit is part of the Network of Exellence 

“BrainNet” Europe devoted to biobanking, harmonization of assessment tools and standardization of 

diagnostic criteria of neurodegenerative diseases. 

The permanent staff of the Division is composed by 8 MD and two technicians. In addition, the staff 

comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1 technician 

who are committed to the research activities on degenerative dementias with different expertise and roles. 

On the overall, the team has large experience in a variety of techniques spanning from neuropathology 

(including immunohistochemistry, morphometry, electron microscopy and atomic force microscopy), to 

molecular genetics, biochemistry (purification and characterization of disease-specific proteins), cellular 

and molecular biology, and animal models. 

Research lines on Alzheimer’s disease 

A. Clinical research 

• Early diagnosis of Alzheimer’s disease (AD) 

We will characterize the phenotypic expression of AD and other degenerative dementias through 

multiplex analysis of a number of variables including neuropsychological and behavioural profile, 

neuroimaging, neurophysiological changes with special attention to sleep abnormalities, genetics, 

and plasma and CSF biomarkers. Furthermore, we will carry out longitudinal studies of 

presymptomatic carriers of gene mutations to detect early markers of conversion to the disease 

state. 

• Clinical trials 

We will carry out phase II and phase III clinical trials to assess the effectiveness of potentially 

disease-modifying drugs, including molecules targeted to Aβ and protein tau. 

• Identification of disease-specific biomarkers 

We are currently working on a peripheral marker of AD having high specificity and sensitivity, 

based on cyclic amplification of misfolded Aβ and Aβ oligomers from biological fluids, similar to 

the PMCA technique successfully developed by Claudio Soto for prion diseases. The same 

approach will be used to amplify and detect misfolded tau. 

• Biobanking and neuropathological and molecular characterization of patients

We will collect systematically plasma, DNA and CSF from patients with different types of 

dementia and non-demented individuals, and perform autopsies for neuropathological 

characterization and biochemical and molecular studies. 

B. Preclinical research 

• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic 

strategy for sporadic AD 

We have recently identified an APP mutation (A673V) that causes early-onset AD only in the homozygous 

state while the heterozygous carriers are not affected. This mutation strongly boosts the production and 

amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and wild-type peptides 

inhibits amyloidogenesis and Aβ-mediated neurotoxicity (Di Fede et al., Science 2009, 323:1473-7). These 

findings are consistent with the observation that the A673V heterozygous carriers do not develop disease 

and offer grounds for a novel therapeutic strategy based on modified Aβ peptides. 

A research priority of our lab is to unravel the molecular mechanisms of the opposite effects of the 

A673V APP mutation in homo- or heterozygous state on amyloidogenesis and to develop a lead 

compound for AD therapy based on A673V-modified Aβ peptides or peptido-mimetic molecules. 

To accomplish these objectives we have generated a panel of transfected cells and transgenic C. 

elegans expressing human APP or Aβ with the A673V mutation, respectively, and transgenic 

mouse lines expressing A673V-mutated APP in the homozygous or heterozygous state. 

Furthermore, we have identified a prototypic lead compound corresponding to a six-mer Aβ peptide 

with the A673V substitution and are currently working on brain delivery systems. 

• Pathways leading to tau pathology 

A key event in AD pathogenesis is the hyperphosphorylation, misfolding and aggregation of the 

microtubule-associated protein tau leading to formation of neurofibrillary tangles, disruption of the 

neuronal cytoskeleton and neurodegeneration. Following the “Aβ cascade hypothesis” this event is 

triggered by aggregated Aβ species, particularly oligomeric assembly intermediates. However, the 

pathways linking Aβ and tau pathology are unknown. This is largely due to lack of animal models able to 

reproduce these two central aspects of AD. We have developed a mouse model of prion disease that shows 

a secondary tauopathy following deposition of PrP amyloid (Giaccone et al., Neurobiol. Aging 2008, 

29:1864-73). This model will be used to investigate the molecular basis of tau pathology induced by 

deposition of an amyloid protein. 

• Role of nuclear tau in neurodegeneration in fronto-temporal dementia 

Tau is the major microtubule-associated protein of neurons and its primary role is to promote assembly and 

stabilization of microtubules required for morphogenesis and axonal transport. We have recently found that 

tau plays an important role also in chromosome stability, and mutations in the tau gene cause chromosome 

aberrations in peripheral cells in addition to formation of neurofibrillary tangles in neurons and glial cells. 

Our objective is to elucidate the molecular mechanisms underlying the genomic instability due to tau 

mutations using cellular models, and to investigate the contribution of the aneuploidy caused by mutated tau 

to neurodegeneration using a transgenic mouse model of tauopathy.

Relevant ongoing research support 

• Cariplo Foundation (01/01/07-30/06/10) 

Title: Genoproteomics of Age Related Disorders (GuARD). 

PI: Roberto Sitia 

Role: Project leader 

• Italian Ministry of Health (01/01/09-30/06/11) 

Title: Alzheimer's disease: development and validation of a multi-factorial protocol for the diagnosis and 

follow-up of disease in the prodromal and incipient phase. 

PI: Fabrizio Tagliavini 

• European Union/Italian Ministry of Health (01/02/09-31/01/12) 

Title: Molecular mechanisms underlying synaptic dysfunction in prototypic neurodegenerative diseases 

related to protein misfolding (nEUROsyn). 

Role: PI 

• Italian Agency of Drug (01/09/06-31/08/10) 

Title: A randomized, double-blind pilot study versus placebo for the evaluation of the efficacy of 

doxycycline administered by oral route in patients affected by Creutzfeld-Jakob disease. 

Role: PI

Nome Roberta Ricciarelli, Ph.D. 

Contatti 

E-mail: ricciarelli@medicina.unige.it 

Tel.: +39-010-3538838; Fax: +39-010-3538836 

Istituto/Dipartimento Section of General Pathology 

Department of Experimental Medicine (DiMeS) 

University of Genoa, Italy 


Dr. Roberta Ricciarelli’s ongoing research on Alzheimer’s disease: 

• Cholesterol and amyloid-beta: is there a relation? 

• Study of the nonmuscle myosin II involvement in the processing of the amyloid precursor protein (APP) 

• Specific type 4 phosphodiesterase inhibitors and their impact on the production of amyloid-beta 



-Basic research, 

-Multidisciplinary projects, 

-New treatment strategies. 

Titolo progetto Molecular mechanisms of N-truncated amyloid beta peptides 

generation 

Ente finanziatore MIUR (Ministry of Education, University and Research) 

Durata progetto Two years (2004-2005) 

Abstract del progetto The amyloid precursor protein (APP) is an integral membrane protein 

processed by several different proteases called secretases. β-Secretase is 

responsible of the N-terminal cleavage of amyloid-� (Aβ). The gene for βsecretase 

(also referred to as BACE) is located on chromosome 11, but no 

AD-related mutation in this gene has been identified so far. BACE mRNA is 

highly expressed in the brain and is also found in a variety of human 

tissues, consistent with the fact that Aβ is normally produced by many 

cell types. Recently, three new alternatively spliced transcripts of human 

BACE have been described, but the functional importance of the different 

BACE isoforms, and their implication in the pathogenesis of AD, has not 

yet been investigated. Our preliminary sequence analysis indicates that, 

as a cause of the alternative splicing, the DTG sequence, which is 

conserved at the active site of the enzyme, may shift to a different 

position within the catalytic site, affecting its substrate specificity. Since 

BACE is involved in the generation of the N-terminus of Aβ species, the 

existence of four BACE isoenzymes prompted us to investigate the 

possible correlation between the expression of selective BACE isoforms 

and the formation of neurotoxic N-truncated Aβ peptides. This

hypothesis is in line with the reported absence of AD-causing mutation in 

BACE gene. 

Titolo progetto In vitro effect of PPAR-γ2 Pro12Ala polymorphism on the deposition of 

Alzheimer's amyloid-β peptides 

Ente finanziatore Fondazione CARIGE 

Durata progetto One year (2007) 

Abstract del progetto Mounting evidence suggests that peroxisome proliferator-activated 

receptor-γ (PPAR-γ) is involved in the modulation of pathogenic events 

related to Alzheimer's disease (AD). Such events would include the 

cerebral deposition of amyloid-β (Aβ) and the consequent local 

inflammatory response. PPAR-γ has been shown to act on both fronts, 

reducing either the secretion of Aβ or the expression of pro-inflammatory 

cytokines. Recently, the relatively common Pro12Ala polymorphism in 

exon 2 of PPAR-γ has been associated with higher risk for late onset AD. 

The aim of the present project is to assess the impact of PPAR-γ and its 

genetic variant on the secretion of Aβ in neuronal cultured cells. 

Titolo progetto Study of physiological function of the amyloid precursor protein 

Ente finanziatore University of Genoa 

Durata progetto One year (2008) 

Abstract del progetto The Alzheimer’s disease (AD) brain pathology is characterized by 

extracellular deposits of amyloid-� (A��) peptides and intraneuronal 

fibrillar structures. These pathological features may be functionally 

linked, but the mechanism by which A�� accumulation relates to 

neuronal degeneration is still poorly understood. A�� peptides are 

fragments cleaved from the amyloid precursor protein (APP), a 

transmembrane protein ubiquitously expressed in the nervous system. 

Although the proteolytic processing of APP has been implicated in AD, the 

physiological function of APP and the subcellular site of APP cleavages 

remain unknown. The overall structure of the protein and its fast 

anterograde transport along the axon support the idea that APP functions 

as a vesicular receptor for cytoskeletal motor proteins. In the current 

study, we will test the hypothesis that myosin II, important contributor to 

the cytoskeleton of neuronal cells, may influence the trafficking and/or 

the processing of APP. Our preliminary results demonstrated that 

downregulation of myosin II-B, the major myosin isoform in neurons, is 

able to increase A�� deposition, concomitantly altering the subcellular 

localization of APP. These new insights might be important for the 

understanding of the function of APP and provide a novel conceptual 

framework in which to analyze its pathological role.

Nome Patrizia Guarneri 

Contatti guarneri@ibim.cnr.it 

Istituto/Dipartimento Istituto di Biomedicina e Immunologia Molecolare – Dip- di Medicina - 

CNR- Palermo 


Rare neurodegenerative diseases: basic research 

Area di interesse identificata Rare diseases 



Pathogenesis of the CLN8 form of the late-infantile neuronal ceroid 

lipofuscinosis 


1) MIUR “PRIN” Programmi di Ricerca Scientifica di Rilevante Interesse 

Nazionale (DM n. 1407 del 4 dicembre 2008). “IDENTIFICATION OF 

BIOLOGICAL, GENETIC AND CLINICAL MODIFIERS OF THE RATE OF 

PROGRESSION AND SURVIVAL IN AMYOTROPHIC LATERAL SCLEROSIS ”, 

prot. 20082XH2Z4-001 



2) FIRB-MIUR, n. RBAU01E3SL, “RETINAL DEGENERATION IN MND 

MOUSE, AN ANIMAL MODEL OF NEURONAL CEROID LIPOFUSCINOSES OR 

BATTEN’S DISEASE. (dec. n. 808/Ric) 

1) Two years 

2) Three years 

The neuronal ceroid lipofuscinoses (NCLs) are a group of 

neurodegenerative, lysosomal diseases with onset usually in childhood, 

being characterized by progressive visual loss, dementia, motor decline, 

therapy resistant epilepsy and early death. Our team has lately focused 

on the pathogenic mechanisms of the CLN8 variant form of the lateinfantile 

NCLs (vLNCL), which is present in Finland, Turkey and Italy. The 

CLN8 gene encodes a transmembrane protein which is located mainly in 

the ER and partially in the ER-Golgi intermediate compartment of both 

neuronal and non-neuronal cells, and it is supposed to have a role in the 

ceramide synthesis. However, its exact function remains undiscovered. 

We have developed a protein-protein interaction study throughout the 

screening of a human brain cDNA library with split-ubiquitin membrane 

two-hybrid system in yeast, which is the only technology developed thus 

far that can work effectively as a in vivo screening system to find 

interactors of ER membrane proteins. This approach along with 

coimmuno-precipitation and -localization assays of transfected cells has 

envisaged for the first time four major protein interactors with the CLN8 

protein. Through in vitro and in vivo studies, we are looking at the 

definition of the functional role of these interactions and particularly of 

the CLN8 interaction with two of the characterized proteins, which are 

known to have functions in synthesis and transport of lipids and are 

suggested to be involved in the amyothrophic lateral sclerosis. In an 

attempt to understand the mechanisms leading to neuronal cell death in 

the disease, we have been also using a spontaneous animal model of the 

CLN8/vNCL, the mnd mouse. We are currently investigating whether and 

how dysfunctions of ER/UPR system and autophagolysosomal pathway

cooperate in the neurodegeneration of the retina and brain structures of 

the mnd model.

Nome Dario Finazzi 

Contatti 

finazzi@med.unibs.it 

phone and fax ++39 030 302356 

Istituto/Dipartimento Dip. Materno Infantile e Tecnologie Biomediche 

Università degli studi di Brescia 








1) Basic research 

2) Genetic susceptibility to Alzheimer’s disease and genome wide 

association studies 

3) Biobanking 

1) IRON METABOLISM IN NEURAL CELLS AND IN 

NEURODEGENERATIVE DISORDERS 

2) ANALYSIS OF CANDIDATE GENES OF SUSCEPTIBILITY TO AD. 

University of Brescia 

Fondazione RSA Agostoni Lissone 

3 years 

Our group is interested in the study of iron metabolism in the brain and 

its possible role in neurodegeneration. The accumulation of iron is a 

phenomenon often detected in the brain from patients affected by 

Alzheimer’s disease and by other neurodegenerative processes. Even 

though such accumulation is not the primary cause of neural death, it is 

surely involved in the progression and deterioration of the 

neurodegenerative process. The excess of iron can be directly linked to an 

increase in the production of free radicals, leading to oxidative damage to 

proteins and membranes, signs commonly documented in tissues with 

neurodegeneration. We aim to a better comprehension of such processes 

by developing two different and complementary lines of research: 1) 

Analysis of the biological mechanisms involved in the maintenance of iron 

homeostasis in the brain, in normal and pathological conditions; 2) 

Analysis of genes linked to iron metabolism as possible modifiers of the 

individual risk to develop AD. 

We intend to perform a thorough analysis of the regulation of iron 

metabolism in neural cells and to assess the way its malfunction may lead 

to iron accumulation. The experiments will be initially carried out in SH- 

5YSY cells (either differentiated or not) and in primary hippocampal 

neurons with the goal to characterize the role played by the hepcidinferroportin 

axis in iron homeostasis of neural cells. We will determine the 

mechanism regulating ferroportin expression and function and describe 

the molecular machinery controlling hepcidin production in neuronal

cells and the role of the peptide in neuronal iron trafficking. The analysis 

will be then extended to wild type mice and murine model of AD 

(APP/PS1dE9), and eventually to autopsy sections of human brain from 

patients and controls. In particular we will investigate the relationship 

between brain inflammation and the regulation of the hepcidinferroportin 

axis. We have preliminary evidence that BMP6 and 

Interleukins intervene in the transcriptional regulation of hepcidin and 

could be of relevance in the progressive accumulation of iron in brain. 

The study take advantage of a significant collaboration with Prof Paolo 

Arosio, one of the major expert in the study of cellular iron metabolism. 

The genetic part of the project will investigate the possible association 

between polymorphisms in genes involved in iron homeostasis (hepcidin, 

ferroportin, DMT1, ceruloplasmin) and the individual risk to develop AD. 

Promoter regions, exons and exon-intron boundary regions will be 

scanned for the presence of sequence variations by DHPLC and real time 

genotyping. The study is granted by the availability of a large set of 

genomic DNA samples from patients and controls and by the 

collaboration with research groups and clinicians with expertise in AD and 

genetic studies (prof. Scarpini, Dott. Galimberti, Dott. Dominici). In 2005 

we set up a bank of different biological samples coming from AD patients 

and sex and age matched controls. The biobanking activity is continuing 

thanks to the collaboration with the Fondazione RSA Agostoni in Lissone 

and Dr Roberto Dominici. Our valid collaboration with the group of Prof 

Scarpini at Ospedale Maggiore in Milan allows us the access to a large and 

well characterized set of genomic DNA samples, as a second and different 

population for the confirmation of data obtained in the first set of 

patients and controls.

Nome Carlo Sala 

Contatti 

Tel. 

E mail 

Carlo Sala 

+390250317096 

c.sala@in.cnr.it 

Istituto/Dipartimento CNR Istituto di Neuroscienze 


Area di interesse identificata Neurotoxic stimuli, synaptic dysfunction 






The role of intraneuronal amyloid β(Aβ) peptide on the pathogenesis of 

the Alzheimer disease: functional and proteomic analysis. 

Fondazione CARIPLO 

2 anni 

La malattia di Alzheimer rappresenta la più comune forma di demenza 

per la quale non esiste ancora nessun intervento terapeutico in grado di 

prevenire, curare o ritardare in modo sostanzaile il suo sviluppo. Recenti 

studi hanno dimostaro che l’accumulo intracellulare di Abeta correla con 

la prima comparsa di deficit cognitivi. Questo progetto si pone 

l’obbiettivo di identificare come Abeta intrecelluare interfereisce con la 

funzionalità delle sianpsi neuronali e, con tecniche proteomiche avanzate, 

di individuare le proteine che interagiasono con Abeta. Infine ci 

proponiamo di studiare i meccanismi molecolari alla base della 

formazione di Abeta purificando e caratterizzando la composizione 

proteica delle vescicole di trasporto per APP. I risultati che ci proponiamo 

di ottenre con questo porgetto potranno contribuire alla conoscenza dei 

meccanismi patogenetici dell’AD e potrebbero consentire lo sviluppo di 

nuovi metodi diagnostici e la eventuale messa a punto di protocolli 

terapeutici

Nome Cristina Limatola 

Contatti 

Tel. 

E mail 

+39 06 49690243 +39 347 2924450 

Cristina.limatola@uniroma1.it 

Istituto/Dipartimento Dipartimento di Fisiologia e Farmacologia, Università Sapienza Roma 


Area di interesse identificata Tra le altre linee di ricerca di interesse del laboratorio, siamo da qualche 

anno coinvolti in studi di ricerca sulla sclerosi laterale amiotrofica (SLA). 

In particolare, in collaborazione con un gruppo di neurologi (Sabatelli, 

Neri) e genetisti (Zollino) dell’Università Cattolica di Roma, abbiamo 

descritto, in pazienti affetti da SLA, delle mutazioni a carico di diverse sub 

unità del recettore nicotinico neuronale, nel loop regolatorio 

citoplasmatico (Sabatelli et al., 2009). Queste mutazioni, in buona misura 

a carico di potenziali siti di fosforilazione da parte di protein chinasi, 

determinano delle alterazione funzionali specifiche di alcune sub unità 

per recettori-canale per l’acetilcolina, con una generale “gain of function” 

recettoriale. L’aumentata responsività alla stimolazione con acetilcolina a 

livello dei terminali glutamatergici potrebbe determinare una liberazione 

eccessiva di glutammato eccito tossico, mentre la potenziata attività dei 

recettori nicotinici potrebbe aumentare l’ingresso di calcio intraneuronale 

a livelli tossici, contribuendo all’inizio o alla progressione della malattia. 

Stiamo attualmente continuando questi studi molecolari, analizzando gli 

effetti dell’adenosina e dell’ATP, i cui livelli sono alterati nella risposta 

infiammatoria cronica caratteristica della SLA, sull’attività dei recettori 

nicotinici neuronali mutati individuati rispetto alla controparte normale. 

Altro punto di interesse riguarda il coinvolgimento delle cellule gliali, 

astrociti e microglia, nello sviluppo e nella progressione di questa 

patologia. Abbiamo sviluppato a tal proposito dei sistemi di co-coltura 

motoneuroni-cellule gliali e di fettine ottenute dal tronco cerebrale 

(regione del nucleo ipoglosso) per studiare il contributo della 

componente gliale nella alterazione dell’attività elettrica ritmica, nelle 

proprietà dei recettori per neurotrasmettitori, sensibilità 

all’eccitotossicità da glutammato, e ci proponiamo di applicare questi 

studi a modelli cellulari che overesprimano forme mutate di proteine 

associate alla SLA, come FUS, TDP-43 o SOD (in collaborazione con un 

gruppo di biologi molecolari della nostra università, Prof. Bozzoni). 



Studio funzionale di mutazioni che aumentano la suscettibilità alla 

Sclerosi Laterale Amiotrofica su calciatori amatoriali o professionisti. 

Ente finanziatore Fondazione Milan 


Abstract del progetto La sclerosi laterale amiotrofica (SLA) è una malattia devastante che 

distrugge in maniera selettiva una particolare categoria di cellule nervose: 

i motoneuroni. Questi sono i neuroni che controllano la contrazione dei 

muscoli scheletrici, per cui la loro morte provoca una paralisi progressiva 

dei soggetti affetti da SLA e, nel giro di 2-6 anni dall’insorgenza della 

malattia, la loro morte (solitamente per arresto respiratorio). 

Solo il 5% dei casi di SLA ha chiaramente un’origine ereditaria, nel 

restante 95% la malattia ha un origine incerta (SLAs). Tra i giocatori di 

calcio, caso unico tra tutte le categorie di sportivi professionisti, il rischio 

di sviluppare una SLAs è ben 6 volte superiore a quello della popolazione 

generale. Non è ad oggi nota la ragione di questo fenomeno. Si ritiene

che i principali fattori di rischio per lo sviluppo delle SLAs, oltre all’età e al 

sesso maschile, siano i traumi meccanici, l’esposizione ad erbicidi, 

un’attività fisica molto intensa e il fumo di sigarette. Da un punto di vista 

cellulare, una delle cause che si ritiene essere alla base di questa malattia, 

è il danno prodotto dal massiccio aumento dello ione calcio all’interno del 

neurone. Siccome la stimolazione dei recettori della nicotina (la sostanza 

contenuta nelle sigarette) che sono localizzati sulla membrana dei 

motoneuroni consente l’ingresso del calcio all’interno del motoneurone si 

è pensato di studiare la frequenza delle mutazioni dei geni che codificano 

i recettori nicotinici neuronali. Queste mutazioni sono presenti nel 6% di 

pazienti affetti da SLAs e inducono tutte un’alterazione nel 

funzionamento dei recettori nicotinici neuronali che provoca un aumento 

dell’ingresso del calcio all’interno del motoneurone aumentando la 

probabilità che esso muoia. Data l’anormale frequenza di questa malattia 

nelle persone che praticano l’attività calcistica a livello agonistico 

(amatoriale o professionale), ci proponiamo di valutare, mediante analisi 

del DNA estratto con semplici prelievi di sangue, la presenza di mutazioni 

dei recettori nicotinici in calciatori in buone condizioni di salute e/o di 

coloro che si sono già ammalati di SLAs. Siamo convinti che lo studio delle 

modalità precise delle modificazioni del funzionamento dei recettori 

nicotinici neuronali dovute a queste mutazioni genetiche, possa essere il 

primo e fondamentale passo per sviluppare una terapia innovativa mirata 

a ripristinare il corretto funzionamento dei recettori con la accelerazione 

della loro desensibilizzazione, diminuendo significativamente il rischio 

dell’insorgenza della malattia, cioè la suscettibilità alla malattia. A questo 

scopo ci proponiamo di caratterizzare dal punto di vista funzionale le 

mutazioni trovate in sistemi eterologhi ricostituiti in vitro (linee cellulari 

trasfettate); inoltre intendiamo sviluppare un modello di topo 

transgenico per una delle mutazione dei recettori nicotinici associata a 

SLA, e utilizzarlo come modello in vivo per studiare la fisiologia del 

muscolo e la fisiologia neuronale. 


Molecular and functional approaches to investigate the 

physiopathological role of the chemokines and their receptors in the 

central nervous system. 

Ente finanziatore Fondazione Cenci Bolognetti 


Abstract del progetto Chemokines and their receptors are widely expresses in CNS where they 

play important roles both under physiological and pathological 

conditions. Chemokines are implicated in the control of neuronal and glial 

development, both in terms of migration and maturation, modulate 

synaptic transmission and have trophic and protective effects on neurons 

and glia. The spectrum of biological activities of chemokines is broadened 

by their involvement in the pathogenesis and maintaining of several 

neuropathologies, when their expression may be altered. Aim of the 

proposed research project is to investigate the physiopathological 

activities of chemokines as modulators of neuronal functions, identifying 

the molecular pathways involved. In particular the effects of chemokines 

will be investigated in brain slices, primary neuronal cultures, cell lines 

and expression systems as concern synaptic transmission, short-term 

neuromodulatory activities and long-term plasticity, trying to correlate 

these events to the activation of specific molecular effectors. These 

investigations will be correlated with studies of chemokine-induced

communication between different cell types, like glial cells and neurons. 

The effects of chemokines as modulators of cell migration will be 

investigated in glial cells and neurons freshly isolated from rodents and 

the signal transduction pathways involved will be investigated. The 

activities of chemokines as neuroprotective substances will be 

investigated in vitro in neurotoxicity models and in vivo in animal models 

of cerebral ischemia. Overall, this study aspire to help to improve the 

understanding of neurogenesis and neurorepair, yielding possible clues to 

therapeutical strategies.

1. Exposing tau pathology at its earliest 

IIT DEPARTMENT OF NEUROSCIENCE AND BRAIN TECHNOLOGIES 

ONGOING PROJECTS ON NEURODEGENERATION 

Fabio Benfenati, MD 

Team: Laura Gasparini; Axel Blau, Paolo Medini; in collaboration with M. G. Spillantini, M. Goedert 

Funding: Compagnia di San Paolo and IIT + pending. 

The study is aimed at revealing functional changes typical of initial stages of tauopathy, and elucidating 

the underlying molecular mechanisms. This knowledge will reveal markers suitable for early clinical 

detection of tau-driven degeneration and identify key pathogenetic mechanisms to be targeted by new 

therapies. 

2. Effects of β-amyloid aggregates on neuronal viability and microglial function 

Team: Denise Ferrera, Claudio Canale, Fabio Benfenati, Laura Gasparini 

Funding: IIT + pending 

We have demonstrated that Ab42, but not Ab40, reproducibly aggregates into stable oligomers and 

protofibrils and reduces the viability of mouse hippocampal neurons. We are now investigating how 

distinct Aβ aggregates affect microglia viability and activate microglial neuroinflammatory response 

and the latter response on neuronal physiology. 

3. β-amyloid aggregates and astrocytic function in Alzheimer pathophysiology 

Team: Tommaso Fellin, Laura Gasparini, Nadia Mazzaro 


We will use an interdisciplinary approach including two-photon microscopy, Ca2+ imaging, 

biochemistry and pharmacology to test the hypothesis that b-amyloid (Ab) oligomers induce reactive 

gliosis through the overactivation of astrocytic NMDA receptors and we will evaluate whether the 

NMDA open-channel blocker memantine antagonizes Ab-induced astrocytic reactive response. 

4. Molecular and structural determinants for β-amyloid mitochondrial accumulation and toxicity 

Team: Mounia Chami, Fabio Benfenati, Dolores Del Prete 

Funding: IIT + pending

Mitochondrial dysfunction is emerging to play a pivotal role in Alzheimer’s disease (AD). Our project is 

aimed at: (i) studying ER-mitochondria communication and APP and Aβ distribution in AD models; (ii) 

modulating ER-mitochondria communication by pharmacological (Ca2+-drugs) and genetic/molecular 

biology through expression of mitochondrial fission and fusion proteins and/or S1T and consequently 

analyse mitochondrial function, and the intracellular distribution of APP and Aβ; (iii) determining the 

APP/Aβ partners in the MAM fraction using a proteomic approach. 

5. Calcium modulating drugs as a therapeutic tool for Alzheimer’s disease 

Team: Mounia Chami, Fabio Benfenati, Dolores Del Prete 


The ability of neurons to regulate Ca2+ signals is compromised during aging. Importantly, alteration of 

Ca2+ regulating systems has been shown to occur early in Alzheimer’s disease (AD). We will 

investigate the potential use of Ca2+ drugs as a mean to reduce Aβ production in different AD models 

in vitro, analyze the potential effect of Ca2+ drugs on beta and gamma secretase activity and apply Ca2+ 

drugs on animal models in vivo and consequently analyze their effect on Aβ plaque formation, synaptic 

activity and cognitive development and/or progression. 

6. Pathogenesis and therapeutic targets for SBMA 

Team: Mara Pennuto, Fabio Benfenati, Chiara Scaramuzzino 

Funding: MDA, Marie Curie, IIT + pending 

Polyglutamine diseases belong to the broad family of brain folding diseases. We aim at using Spinobulbar 

muscular atrophy (SBMA) as a model of polyglutamine disease to identify signaling pathways 

that impact brain folding diseases. Recent evidence shows that the IGF-1/Akt signaling pathway impact 

brain folding diseases. However, the mechanisms underlying the effect of Akt signalling remain to be 

elucidated. By putting polyglutamine AR downstream of Akt with our previous research, we 

established SBMA as a good model to tease apart the molecular details of the effect of IGF-1/Akt in 

polyglutamine disease. 

7. Interactions of Aβ and α-synuclein with the cytoskeleton 

Team: Evelina Chieregatti, Giuseppe Ronzitti, Anako Tsushima 

Funding: IIT, Cariplo Foundation + pending

Both Aβ and α-synuclein are implicated in actin dynamics as well as microtubule stability through 

their interactions with a microtubule-binding protein, tau. Our projects concern: (i) the study of their 

effects on the morphology and dynamics of actin microfilaments and intermediate filaments; (ii) the 

possible effect of the two proteins on cytoskeletal-regulated processes, such as polarization, growth 

cone formation and regeneration after axotomy; (iii) the identification of proteins of interest such as 

enzymes or molecular motors that may be found in the APP-containing endocytic vesicles. 

8. Mechanisms of neurodegeneration and enhancement of adult neurogenesis to improve 

cognitive functions in Down syndrome 

Team: Andrea Contestabile, Diego Ghezzi, Fabio Benfenati, Laura Gasparini 

Funding: Lejeune Foundation + IIT + pending 

Down syndrome (DS) patients develop neuropathological lesions typical of Alzheimer disease (AD) by 

the fourth decade of life with cerebral accumulation of amyloid-beta (Ab) deposits and intraneuronal 

inclusions made of hyperphosphorylated tau protein (neurofibrillary tangles, NFTs). Mice models of 

DS recapitulate many aspects of the pathology, including: APP-dependent cholinergic dysfunctions 

during aging and tau hyperphosphorylation. This research line aims to better characterize the 

neurodegeneration process driven by APP overexpression and tau hyperphosphorylation in a DS mouse 

model and evaluate potential therapeutic interventions to alleviate the pathology. Down syndrome is 

also characterized by decreased neurogenesis both during development and adulthood. Adult 

neurogenesis in the DG of rodents has been associated to memory formation and hippocampaldependent 

learning. This research line aims to elucidate the impact of adult neurogenesis on learning 

and memory in a mouse model of DS and assess the effect of pharmacological treatments able to 

enhance the neurogenesis process.

Nome Armando Genazzani 

Contatti 

Tel. 

E mail 

DiSCAFF, Via Bovio 6 - Università del Piemonte Orientale, Novara 

0321-375827 

genazzani@pharm.unipmn.it 

Istituto/Dipartimento DiSCAFF, Università del Piemonte Orientale 


Area di interesse identificata Malattia di Alzheimer e trasduzione del segnale legata al calcio 



Role of calcium-dependent gene expression and protein trafficking in 

shaping post-synaptic activity: implications for neurodegenerative 

diseases 

Ente finanziatore Fondazione Cariplo 

Durata progetto 2 anni 

Abstract del progetto Alzheimer disease (AD) is a progressive neurodegenerative disease 

characterized by a progressive cognitive and memory decline, increasingly 

frequent with advancing age until it affects as many as 50% of individuals 

85 years of age and older. There is no remission in the progression of the 

disease nor are there any truly effective pharmacological interventions 

available at present. In the framework of an ageing society a better 

understanding of pathogenic mechanisms and identification of new 

pharmacological targets are key strategies. 

For many years, AD was attributed to neuronal death induced by deposits 

of fibrillar Amyloid b. The production of amyloid ß (Aß) is mediated by the 

concerted action of two different secretases, namely ß-secretase (BACE) 

and gamma-secretase, showing a proteolytic action on the amyloid 

precursor protein (APP). Alternatively, APP can be subjected to the 

proteolytic cleavage by a--secretase (ADAM10), which occurs within the 

sequence of Aß, thus precluding the formation of the amyloidogenic 

fragments. Yet, a recent view suggests that Abeta production has an 

impact primarily on the post synaptic compartment of the excitatory 

glutamatergic synapse (this preceding cell death and correlating with 

clinical features of the disease), as demonstrated by in vitro studies, thus 

indicating Alzheimer disease as a synaptopathy. 

An additional biochemical parameter which has been linked to both 

synaptic plasticity events and neurodegeneration is Calcium. The link 

between calcium and Alzheimer has been strengthened by a number of 

independent groups. Calcium dyshomeostasis appears to be an integral 

part of the pathology, either because it can influence Abeta production or 

because it can be altered by amyloid beta overproduction. 

This project will focus primarily in dissecting the contribution of calcium 

dyshomeostasis to the AD synaptopathy. Thus, the main goal of the 

project is to investigate the link between dys-regulation of calcium 

signalling and early stages of AD – both at cellular and molecular level - 

working on the hypothesis that this neurodegenerative disorder is a 

synaptopathy. In particular we will firstly characterize calcium dependent 

processes in an innovative animal model of Alzheimer disease obtained 

by uncoupling ADAM10 from its cargo protein SAP97 which regulates 

enzyme activity. Secondly, we will address the role of calcium in both 

trafficking and gene expression of key synaptic proteins in vitro and in in 

vivo models of altered amyloid cascade.

The investigation of these objectives would provide novel insights in the 

understanding of the molecular basis of Alzheimer’s disease in early 

stages and would possibly contribute in identifying new targets for 

therapeutic intervention

Nome Dr. Andrea Contestabile 

Contatti 

Email: andrea.contestabile@iit.it 

Phone: ++39-010-71781785 

FAX: ++39-010-71781230 

Istituto/Dipartimento The Italian Institute of Technology (IIT) 

Department of Neuroscience and Brain Technologies 

Via Morego 30, 16163 Genova, Italy 


Area di interesse Down syndrome (DS) or trisomy 21 is the leading cause of genetically-defined 

identificata mental retardation and is characterized by decreased cognitive functions, 

developmental defects, impaired neurogenesis and Alzheimer (AD)-like 

neurodegeneration during aging. The progression of Alzheimer neuropathology in 

DS individuals appears to be analogous to sporadic cases of AD with cerebral 

accumulation of amyloid-beta (A�) deposits (i.e., A� plaques) and intraneuronal 

inclusions made of hyperphosphorylated tau protein (i.e., neurofibrillary tangles, 

NFTs). However, its appearance is anticipated by at least twenty-thirty years with 

respect to the general population. The presence of an extra-copy of the 

A��precursor protein (APP) gene, located on human chromosome 21 (HSA21), is 

believed to be directly responsible for early deposition of A� in DS brain. The gene 

encoding Dyrk1A is also located in the triplicated region of HSA21 and is 

overexpressed in DS brains. Recently it has been proposed that Dyrk1A may be 

involved in tau hyperphosphorylation and NFTs formation typical of DS and AD. 

Dyrk1A phosphorylates tau at several sites, and such sites are 

hyperphosphorylated in adult DS brains. In addition, phosphorylation by Dyrk1A 

primes further phosphorylation of tau by Glycogen Synthase Kinase-3� (GSK-3�� 

suggesting that the extra copy of the Dyrk1A gene on HSA21 contributes to the 

early onset of AD neuropathology seen in DS. In fact, a role for Dyrk1A in AD 

neurodegeneration has been recently identified, therefore suggesting a functional 

link between some of the triplicated genes found in Down syndrome and Alzheimer 

disease. 

The Ts65Dn mouse is the best characterized animal model of DS and carries an 

extra copy of the distal segment of mouse chromosome 16 which is synthenic to 

human chromosome 21. The Ts65Dn mouse recapitulates many aspects of DS, 

including: APP-dependent cholinergic dysfunctions during aging; Dyrk1A-mediated 

tau hyperphosphorylation and impairment of hippocampus-dependent memory 

function. 

Ts65Dn mice also display reduced adult neurogenesis in the two known adult 

neurogenic niche in mammalian brain, i.e, the subventricular zone (SVZ) of the 

lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus (DG) in the 

hippocampus. Adult neurogenesis has been also found to be reduced in several 

mouse models of AD, further supporting the notion of a functional connection 

between DS and AD pathological mechanisms. Adult neurogenesis in the DG of 

rodents has been associated to memory formation and hippocampal-dependent 

learning such as spatial and contextual recognition. Decreased adult neurogenesis 

observed in DS and AD mouse models is therefore believed to play an important 

role in the impaired performance of these mice in hippocampus-depended 

memory tasks. 

GSK-3� inhibition has been identified has a possible therapeutic target for AD 

neurophatology. Accumulating evidence also indicates that adult neurogenesis is 

negatively regulated by the activity of GSK3� through the inhibition of the �- 

Catenin pathway, and that adult DG neurogenesis can be stimulated by GSK3� 

inhibitors.

In order to identify effective pharmacological treatment to improve learning and 

memory in mouse models of DS that could be translated into human therapy for 

both AD and DS we are currently evaluating the usefulness of GSK-3� inhibitors to 

stimulate adult DG neurogenesis and alleviate tau hyperphosphorylation in Ts65Dn 

mice. 


Titolo progetto Effects of lithium on cognitive functions, neurogenesis and Tau pathology in 

Ts65Dn mice. 

Ente finanziatore Fondation Jerome Lejeune, France 


Abstract del 

progetto 

Compelling studies indicate that impairment of adult neurogenesis and 

degenerative mechanisms driven by hyperphosphorylation of Tau are the main 

determinants in reducing cognitive and memory function in animal models of 

Down syndrome (DS). Indeed, adult neurogenesis is impaired in the dentate gyrus 

of the hippocampus in the Ts65Dn mouse, a DS model reproducing most 

neuropathological and cognitive deficits of the disease. We have recently shown 

that administration of lithium to Ts65Dn mice is able to restore neurogenesis in the 

subventricular zone of the lateral ventricle, another neurogenetic area of the adult 

brain. Lithium is an extensively used mood stabilizer and acts as an inhibitor of 

Glycogen Synthase Kinase-3��(GSK-3�). GSK-3� activity enhances tau 

phosphorylation and is primed by dual-specificity tyrosine phosphorylated and 

regulated kinase 1A (Dyrk1A), which is encoded by a gene triplicated in DS and 

Ts65Dn mice. Aim of the present project is to test whether lithium long-term 

administration ameliorates cognitive performance in Ts65Dn mice by reducing 

accumulation of hyperphosphorylated tau and by stimulating neurogenesis, 

differentiation and integration of newly-born neurons. Lithium (2.4 g/kg of Li2CO3 

in the chow) will be administered to Ts65Dn for 4 or 8 weeks starting at 6 months 

of age. Hippocampus-dependent memory functions will be evaluated through 

novel object recognition, spontaneous T-maze alternation and geometric task tests 

performed at the end of lithium treatment. After the treatment, adult 

neurogenesis in the dentate gyrus of Ts65Dn mice will be assessed by BrdU labeling 

and evaluation of proliferation markers. The expression of neuronal differentiation 

markers will be analyzed to assess the maturation of newly generated neurons. The 

accumulation of hyperphosphorylated Tau will be quantitatively monitored by 

immunohistochemical and biochemical techniques. Our experimental approach will 

allow testing a novel therapeutic approach for DS by simultaneously targeting two 

known pathological features affecting cognitive functions.

Nome LAURA GASPARINI 

Contatti 

Laura.gasparini@iit.it 

Istituto/Dipartimento Fondazione Istituto Italiano di Tecnologia, Dipartimento di Neuroscienze 

e Neurotecnologia, Via Morego 30, 16163 Genova 





The lab focuses on studying key neuropathological molecular mechanisms 

of neurodegenerative diseases, with the view of understanding the 

disease processes and to translate basic knowledge into potential novel 

therapies. 

Tau-driven degeneration and neurotrophic response: searching disease 

markers for drug screening. Accumulation of intraneuronal incusions 

made of tau protein is a pathological hallmark of the brain of patients 

with tauopathies, including frontotemporal dementia and parkinsonism 

linked to chromosome 17, parasupranuclear palsy, corticobasal 

degeneration and Alzheimer Disease (AD). Our research is aimed at 

providing insight on alterations of neuronal activity associated with the 

onset and progression of tau pathology in transgenic disease models of 

tauopathy in vitro and in vivo, to reveal key tau-driven pathological 

mechanisms, and identify potential molecular targets for drug discovery. 

Neuroinflammatory mechanisms in Alzheimer disease: role of β-amyloid 

aggregates in microglia activation. Neuroinflammation is one of the 

neuropathological features of Alzheimer Disease (AD) and is characterized 

by activation of inflammatory cells (i.e., astrocytes and microglia) in areas 

of the brain affected by β-amyloid (Aβ) plaques and tau pathology, with 

production of altered levels of inflammatory mediators. Our lab 

investigate the pathogenic mechanisms underlying AD-associated 

neuroinflammatory processes, with the main focus on understanding the 

role on Aβ aggregated species in microglia activation. 

Pathogenic mechanisms driven by laminB1 overexpression in adult 

onset leukodystrophy. Lamin B1 (LMNB1) is a component of the nuclear 

lamina. It has been recently discovered that the presence of an extra copy 

of the gene encoding this protein causes autosomal dominant 

leukodystrophy (ADLD). ADLD is an invariably fatal disease characterized 

by white matter loss, movement disorders and dysautonomia. The 

pathogenic mechanisms by which LMNB1 gene duplication leads to 

disease are unknown. Our research aims at clarifying such mechanisms 

using in vitro and in vivo experimental models. 

Tau-driven degeneration and neurotrophic response: searching disease 

markers for drug screening 

Ente finanziatore Compagnia di San Paolo 



Background: Neurofibrillary tangles (NFTs) are neuropathological 

hallmarks of human tauopathies. Reduced levels of neurotrophin TrkB 

receptor have been found in NFT-bearing neurons. Rationale and 

objectives: Initial findings showed that retinal ganglion cells (RGCs) of 

human P301S tau transgenic (TG) mice develop NFTs and have impaired

axon outgrowth upon stimulation with neurotrophins (CNTF/BDNF). We 

propose to investigate the mechanisms by which tauopathy affects 

neurotrophin signalling and neuronal activity to identify disease markers 

and provide a test-bed for evaluating potential neuroprotective drugs in 

vitro. Expected results and relevance: Reveal characteristic signatures of 

tau pathology, suitable to investigate potential tau-targeting 

therapeutics. 


Clinical, neuroradiological and molecular investigation of Adult-onset 

Autosomal Dominant LeukoDystrophy (ADLD): dissection of Lamin B1mediated 

pathophysiological mechanisms in cellular and mouse models. 

Ente finanziator TELETHON 


Abstract del progetto Overall objectives: To clarify Lamin B1 (LMNB1)-mediated pathogenic 

mechanisms of adult-onset autosomal dominant leukodystrophy (ADLD) 

and provide tools to study future therapies. Background/Rationale: ADLD 

is a rare, progressive and fatal genetic disease generally characterized by 

autonomic failure at onset. We have collected two ADLD families with 

LMNB1 gene duplication, and a third with a variant ADLD and an 

unknown mutation affecting LMNB1 expression. Expected results: The 

project will enhance our knowledge on genetic, clinical and 

neuroradiological features of ADLD. It will shed light on its 

pathophysiology and on LMNB1-mediated pathogenic mechanisms. It will 

generate an ADLD mouse model and provide tools (i.e., clinical 

biomarkers, expression constructs, gene-reporter assays, cellular models) 

which will be key elements for further studies of the disease and its 

therapy.

Nome Michele Mazzanti 

Contatti 

michele.mazzanti@unimi.it 

+39 2 50314958 +39 2 50314959 

Istituto/Dipartimento Dept. of Bimolecular Science and Biotechnology 

Proposta di ricerca Investigation on early markers during neurodegenerative process 


D. Oxidative Stress and neurodegeneration 






Beta-amyloid effect on retina cells. 

Investigation on Chloride Intracellular Channel 1 (CLIC1) protein modulation to use as an 

early marker during neurodegenerative process induced by amyloid compounds. 

Merz Biotech, Germany 

Italian Research Ministery 

1 year 

2 years 

One of the major apprehensions of contemporary society is population aging. 

Paradoxically, scientific discoveries applied to medicine have significantly increased the 

life expectation. However, older populations create new medical problems that, once 

episodic, are now common in a growing number of aging individuals. 

Neurodegenerative diseases play a predominant role in this new scenario. A major 

neurodegeneration disorder is Alzheimer Disease (AD), but this is poorly understood 

and not only we are without a successful therapy but also a proper identification of the 

disease in living humans is, at the moment, problematic. The appropriate diagnosis of 

an AD patient can be obtained only from histological analysis of post-mortem brain 

tissue. AD is characterized by an accumulation in neuritic plaques of beta-amyloid (A�) 

protein, infiltrated by reactive microglia. Although A� and in particular its oligomeric 

forms has been shown to exert a toxic effect on neurons, the role of microglia in 

neuronal injury remains unclear. Microglia activation is likely to be a key element in the 

pathophysiology of the local inflammation that accompanies AD. Activation of microglia 

by A� protein triggers the production of toxic free radicals and pro inflammatory 

cytokines which are responsible for the neuronal degeneration. Previous studies 

demonstrated that A� protein stimulation of neonatal rat microglia specifically lead to 

the increase of Chloride Intracellular Channel 1 (CLIC1) expression after 48 hours of 

stimulation. This was not the case when microglia was activated by phorbol myristate 

acid (PMA), Lipopolysaccharide, colony stimulating factor or basic fibroblast growing 

factor, thus quoting CLIC1 as specifically up-regulated by amyloid compounds. The 

cellular and molecular characterization of the protein has revealed that CLIC1 is mainly 

cytoplasmatic and is able to shuttle between the aqueous and the membrane phase 

depending on the activation state of the cell. In the membrane, the protein acts as a 

chloride conductance, allowing ionic flow to compensate the negative charges extruded 

by the membrane NADPH oxidase during reactive oxygen species (ROS) production. We 

have recently showed that suppression of CLIC1 functional expression using (i) inhibition 

of the CLIC1 current, (ii) replacement of extracellular Cl - with impermeant anions, (iii) 

transfection of microglia cells with specific siRNA, or (iv) addition of an antibody against 

the channel protein, significantly reduced Aβ-induced ROS production. In a triple 

mutant Alzheimer mouse model, CLIC1 protein is up-regulated and show a predominant 

membrane localization in microglia cells of cerebral cortex. Furthermore, in human 

brains of Alzheimer patients, CLIC1, like several other proteins, is drastically up 

regulated. To explore the idea that CLIC1 protein could work as a therapeutic target as 

well as specific early marker for amyloid based diseases, we will investigate its 

distribution and modulation in two other neurodegenerative processes. Prion Protein 

syndrome (PrPsc) and Spinocerebellar Ataxia 1 (SCA-1) share with AD the predominant 

�-sheet structure of the presumed pathological protein. As typical amyloid compounds, 

A�, PrP and Ataxin-1 are characterized by insoluble deposits. However, while the action 

mechanism of the first two takes place in the extracellular environment, Ataxin-1 

accumulates in the cell nucleus.

The goal of our study is to pursue CLIC1 as a molecular target for early diagnosis and 

possibly for a therapy of amyloid induced neurodegeneration. Since its specific reaction 

to amyloid peptides, CLIC1 could represent an unambiguous marker of several amyloid 

based neurodegenerative process. It is reasonable to think that its marked increase in 

the brain immune system occurs in the early stage of the pathological process following 

a chronic state of oxidative stress induced by the different amyloids accumulation. We 

will use two different strategies to assess this possibility. It is well known that during 

neurodegenerative states the central nervous system (CNS) is colonized by systemic 

macrophages recruited from the blood stream. It is likely that the up regulation of CLIC1 

protein could be a general feature of all macrophages. This will be tested both in mouse 

models of AD, PrPsc and SCA-1 as well as in blood samples from patients. Alternatively, 

we will explore the possibility to detect CLIC1 increment in the most accessible part of 

the CNS: the retina. Activation of the CNS immune system and neurodegenerative 

process were described in the eyes of AD, PrPsc and SCA-1 patients. Degeneration of 

retinal ganglion cells (RGCs) follows the same pathways of neuronal cell death in 

neurodegenerative diseases. Thus, CLIC1 protein increase should be easily detected in 

the retina immunocompetent cells system, following a chronic condition of oxidative 

stress that is belive to anticipate RGC apoptosis.


C. Neuro Inflammation

Nome GABRIELA CONSTANTIN, M.D., Ph.D. 

Assistant Professor of Pathology 

Contatti 

Email: gabriela.constantin@univr.it 

Tel: 39-045 8027102; 8027120; Fax: 39-0458027127 

Istituto/Dipartimento Department of Pathology, University of Verona, Strada le Grazie 8, 

37134, Italy 



Role of Inflammation mechanisms in neurodegenerative diseases 

In recent years growing evidence show that blood-brain barrier (BBB) breakdown and inflammatory 

responses, may initiate and/or contribute to a ‘‘vicious circle’’ of the disease process, resulting in 

progressive synaptic and neuronal dysfunction and loss in disorders such as Alzheimer’s disease (AD), 

Parkinson’s disease, amyotrophic lateral sclerosis, multiple sclerosis, and others (Zlokovic, Neuron 2008). 

In fact, it is clearly emerging that a better understanding of AD inflammatory and immunoregulatory 

processes may lead to the development of anti-inflammatory approaches that may help to slow-down or 

block the progression of this devastating disorder. 

Our group has established expertise in neuroinflammation/neuroimmunology field and will perform an 

interdisciplinary study to explore the role of vascular inflammation mechanisms and the role of 

leukocytes and leukocyte trafficking in the pathogenesis of AD. 

The following main research lines will be developed: 

1. Characterization of vascular inflammation and leukocyte-endothelial interactions in animal models of 

AD. Our team has previously shown that leukocyte adhesion to brain endothelium leads to BBB damage 

and contribute to neuronal loss (Fabene et al., Nat. Med. 2008). We will first study BBB leakage and 

vascular inflammation by identifying adhesion mechanisms able to mediate leukocyte-endothelial 

interactions in brain vessels. Then we will study the selectins, mucins, integrins and chemokines involved 

in rolling and firm arrest of leukocyte subpopulations such as neutrophils, monocytes, Th1 and Th17 cells, 

which were able to interact with endothelium in brain microcirculation of AD mice in preliminary studies 

performed by our team. To this aim, we will use our established expertise in: in vivo staining, in vivo 

Imaging System IVIS ® 200 (Caliper Life Sciences) and intravital microscopy in cortical microcirculation 

(Constantin et al., Immunity, 2000; Battistini et al., Blood 2003; Pluchino et al., Nature 2005; Fabene et al., 

Nat. Med. 2008; Bolomini-Vittori et al., Nat. Immunol., 2009; Deban et al., Nat. Immunol., in press). Finally, 

we will study the effect of adhesion mechanism blockade whether by inhibitory antibodies or genetic 

deficiency on the induction and evolution of AD and will identify new potential therapeutic targets for AD. 

2. Study of neuro-imuno interactions by using two-photon laser scaning microscopy (TPLSM) in AD 

cortex. Leukocyte motility behavior inside the cortex of transgenic animals with AD will be studied in 

relation to the vicinity of blood vessels, amyloid deposition and presence of neurofibrillary tangles. Taking 

into account our preliminary confocal microscopy studies showing that monocyte, Th1 and Th17 cells 

migrate into the cortex of AD mice, we will study the cell-cell contacts between immune cells and neural 

cells using transgenic mice expressing fluorescent neurons or glial cell populations. This research line will 

be developed based on our TPLSM expertise (Rossi et al., submitted) and our in vivo confocal microscopy 

facility, which consists of a customized upright Leica TCS SP5 AOBS system formed by: microscope DM6000 

equipped with a complete set of PlanApo objectives, ultrafast scanning head SP5 AOBS coupled to Ar-

HeNe sources and a mode-locked Ti:Sapphire Chameleon Ultra II laser (Coherent Inc). 

Overall these studies represent a novel and original contribution to the understanding of AD pathogenesis 

and have the potential to identify new therapeutic targets in AD. 


Titolo progetto Leukocyte recruitment: molecular mechanisms and pathological aspects 


Fondazione Cariverona 


February 1, 2009 - January 30, 2011 

Abstract del progetto The research activity includes 3 tasks: Task 1. Mechanisms involved in the 

control of granulocyte and monocyte recruitment by Src tyrosin kinases; 

Task 2. Role of Rap1, cdc42 and PIP5KC in the control of the modalities of 

integrin activation by chemiotactic factors in T and B lymphocytes; Task 3. 

Role of Src-family tyrosin kinases in the migration of leukocytes in 

inflamed brain vessels and in the induction of neurodegenerative diseases 


Adipose-derived stem cells (ADSC) therapy in experimental autoimmune 

encephalomyelitis 

Ente finanziatore National Multiple Sclerosis Society, New York, USA 

Durata progetto July 1, 2009- June 30, 2011 

Abstract del progetto This project has two objectives. The first objective is focused on the study 

of the clinical and neuropathological effect of ADSC on experimental 

autoimmune encephalomyelitis (EAE), which represents the animal model 

of multiple sclerosis, a neurodegenerative disease. The second objective 

is to study the immunomodulatory mechanisms controlling the effect of 

ADSC on EAE.

Name Marina Bentivoglio, MD, Professor 

Contacts 

Office: +39-045-8027158; fax: +39-045-8027163; e-mail: 

marina.bentivoglio@univr.it 

Istituto/Dipartimento Department of Neurological, Neuropsychological, Morphological and 

Motor Sciences – Medical Faculty – University of Verona 

Strada Le Grazie 8, 37134 Verona, Italy 

Research proposal 

Regional vulnerability of the aging brain to neurodegeneration and immune regulatory mechanisms 

Background and aims: A wealth of evidence in the last years has indicated that normal aging is hallmarked 

by low-grade chronic inflammatory activity, with increased production of proinflammatory cytokines both 

peripherally and in the brain, and decreases of anti-inflammatory mediators. Data from our laboratory 

(e.g. Deng et al 2006; Sadki et al., Neurobiol. Aging, 28, 296-305, 2007. Xu et al., Brain Behav. Immun. 1, 

138-152, 2010) have shown in mice an increase of the immune responsiveness of the aging brain (at the 

molecular and cellular levels, the latter including glial cells and T-cell recruitment). This is in contrast with 

the known peripheral decline of innate immunity during senescence, and could be involved not only in 

cognitive decline but also in the vulnerability of the aging brain to neurodegeneration. On the other hand, 

and importantly, not only aging is the primary risk factor for neurodegenerative diseases, including 

Alzheimer’s disease (AD), but also these diseases are hallmarked by regional vulnerability in the brain, 

which is still elusive. It is, therefore, of primary interest to assess whether immune regional changes in the 

aging brain are implicated in its susceptibility to neurodegeneration. To this purpose, the current proposal 

aims at investigating: i) regional immune-regulatory changes in the aging brain, and ii) the significance of 

such changes in terms of vulnerability of different brain regions to neurodegenerative insults during 

senescence. 

Plan of work: Step 1: Analyses of the following parameters in mice of young age versus an advanced age, 

and in transgenic mice which provide AD models: gene profiling (by means of microarrays and quantitative 

real-time RT-PCR) of immune-regulatory genes (including adhesion molecules, complement, cytokine and 

chemokine receptors, MHC antigens, Toll receptors, etc.), and immunophenotyping of cells (microglia, 

astrocytes, perivascular macrophages, T-cell subtypes, dendritic cells) in different dissected brain regions 

(neocortex, hippocampus, hypothalamus, midbrain, etc); magnetic resonance imaging with ultra-small 

superparamagnetic particles of iron oxide (USPIO) to visualize regional cellular infiltration. Step 2: A: 

Application of challenges with different potentially beneficial or detrimental approaches (proinflammatory 

stimuli, like lipopolysaccharide chronic administration; anti-inflammatory therapy; enriched environment, 

which is known to exert a beneficial effect; chronic sleep restriction -frequent during normal aging and 

known to affect cognitive performance), and test the outcome of these challenges on immune-regulatory 

gene expression, neurodegeneration and behavior/cognitive performance. B: Investigate the eventual 

occurrence of neurodegeneration during aging in mice with targeted deletion of immune molecules (e.g. 

STAT-I signaling pathway knockout, interferon-gamma or its receptor knock-out, etc.) versus wild-type 

littermates. 

Identified area of interest 

Current funding 

Project title 

Prediction of cognitive properties of new drug candidates for 

neurodegenerative diseases in early clinical development (Acronym:

Funding agency European Commission 

Project duration 

01/24/2010- 01/23/2014 

Project abstract 

PHARMA-COG). IMI/115009 - IP (Integrated project) 

(note: murine models of AD are available in the framework of this project) 

Recently the EU Council of Ministers for Health underlined the 

importance of generating novel therapeutic agents both for symptomatic 

and disease modifying treatment of Alzheimer’s disease (AD). However, 

despite the increase in translational medicine activities attrition rates still 

remain high and progress in bringing these biomarkers and models to a 

state of readiness as effective decision making tools is slow as each 

academic and pharmaceutical company work in isolation. 

Bringing together European experts in technologies fully translatable 

from animal to human, experts in translational medicine, drug discovery 

and mathematical modelling, PHARMA-COG proposes to accelerate this 

validation using a ‘MATRIX’ approach i.e. conducting parallel experiments 

in animals and human using a comprehensive and standardised battery of 

behavioural, neurophysiological, morphological/functional imaging, and 

biochemical endpoints to: 

o develop models with greater predictive capacity for the clinics 

o develop and validate translatable pharmacodynamic markers to 

support dose selection 

o develop challenge models to support early hint of efficacy studies 

o identify and validate of markers of disease progression and patient 

stratification.

Nome Michela Matteoli 

Contatti 

Tel. 

E mail 

02 50317097 

Michela.matteoli@unimi.it 

Istituto/Dipartimento Farmacologia Chemioterapia e Tossicologia medica 


Toxic effects of beta-amyloid mediated by microglial cells 

Area di interesse identificata Neurotoxic stimuli 






Nome Elisabetta VEGETO 

Contatti 

Tel. 

E mail 

Via Balzaretti, 9 

02 50318263 

elisabetta.vegeto@unimi.it 

Istituto/Dipartimento Dept. Pharmacological Sciences, University of Milan 


Area di interesse identificata Gender and hormonal/endocrine signals in neuroinflammation: role in 

Alzheimer’s disease and brain aging 

Publications from the laboratory demonstrated a key role for estrogens in 

regulating microglia cells reactivity in an experimental model of 

Alzheimer’s disease and in other neuroinflammatory conditions. 

Accordingly, we showed that hormonal manipulations, achieved through 

experimental models of menopause and estrogen therapy, impinge on 

neuroinflammatory signs in the aging female brain. Considering that 

menopause is associated with a higher risk to develop AD in women and 

that estrogens are consistently associated with neuroprotective effects, 

our research is focused on the identification of the cellular and molecular 

targets of estrogens that modulate the susceptibility to 

neuroinflammation. Since estrogens action is mediated through the 

interaction with specific intracellular receptors, our studies are aimed at 

identifying selective estrogenic drugs and appropriate biomarkers of 

hormone signaling in neuroinflammation. 



Estrogen activity in microglia as a potential pharmacological target for AD 

therapy (GP0127Y01) 

Ente finanziatore Telethon; 126,000 Eu 

Durata progetto 3 yrs (2001-2003) 

Abstract del progetto This study was aimed at evaluating estrogens activity on the activation of 

microglia cells that is evoked by beta-amyloid deposition in the brain of 

APP23 mice, an animal model of AD. Results show that withdrawal of 

circulating estrogens through ovariectomy leads to an increased number 

of amyloid deposits that show reactive microgliosis, whereas estrogens 

replacement reduce this event to control levels. These results support 

the hypothesis that sicrulating estrogens are beneficial signals that acting 

also on the neuroinflammatory component of AD delay pathological signs 

of this disease. 

Titolo progetto Estrogens and Women Ageing (Contract no. 518245) Specific Targeted 

Research or Innovation Project 

Ente finanziatore European Commission; 510,000 Eu 

Durata progetto 3 yrs (2005-2007) 

Abstract del progetto This project investigated on the efficacy of estrogenic drugs in reducing 

the neuroinflammatory response in the brain of aging female mice, with 

particular emphasis on the timing of drug administration in relation with 

menopause induction. Results show that menopause potentiates the 

neuroinflammatory response in brain and that the anti-inflammatory 

activity of several estrogenic compounds is reduced in parallel with 

increasing time of hormone deprivation. These results implicate that 

hormone replacement therapy might benefit from the anti-inflammatory 

activity of estrogenic compounds in brain only when therapy initiates

closer in time with the onset of menopause. 

Titolo progetto MADRI (Menopause: decreased response to increasing inflammation) 5 

R01 AG027713-02 

Ente finanziatore NIH; 200,000 Eu 


Abstract del progetto This project investigates more directly on the effect of menopause and 

aging on the inflammatory response of the brain. Preliminary results 

show that menopause modifies the susceptibility of brain inflammatory 

cells that is already altered by the aging process. 


DIMI (Diagnostic Molecular Imaging ) A Network of Excellence for 

Identification of new imaging markers for diagnostic purpose 

Ente finanziatore European Commission; 376,000 Eu 


Abstract del progetto This project is aimed at challenging imaging techniques for improving 

studies on brain reactivity, including inflammation. Several reporter and 

transgenic animal models were generated under this program; the main 

results from this project show that it is now feasible to trace estrogen 

transcriptional activity in brain 


Role of estrogens in brain vulnerability to neuroinflammatory signals 

associated with aging and neurodegenerative diseases 

Ente finanziatore Ministero della Salute; 20,000 Eu 

Durata progetto 2009 

Abstract del progetto Aim of the study was to analyse the reactivity of macrophage cells from 

male and female mice in response to pathological signals that arise in the 

aging brain.

Nome Dr. Barbara Viviani 

Prof. Marina Marinovich 

Dr. Mariaserena Boraso 

Prof. Corrado L. Galli 

Contatti Tel: +39 (0)2 50318356 

Mail: 

barbara.viviani@unimi.it 

mariaserena.boraso@unimi.it 

marina.marinovich@unimi.it 

corrado.galli@unimi.it 

Istituto/Dipartimento Department of Pharmacological Sciences, University of Milan 

Proposta di ricerca Role of pro- and anti-inflammatory cytokines in the prognosis 

and progression of neurodegenerative diseases 


identificata 






MOLECULAR MECHANISMS OF INFLAMMATION AND REPAIR IN STROKE: 

NEURORADIOLOGY AND BIOCHEMICAL ENDPOINTS TO DESIGN A NOVEL 

THERAPEUTIC AND PROGNOSTIC APPROACH 

Regione Lombardia 

24 mesi 

Stroke remains a leading cause of adult death and disability, which still demand an 

appropriate therapeutical approach. Some degree of spontaneous behavioral recovery is 

usually seen in the weeks after stroke onset. In general, the best outcomes are associated 

with the greatest return toward the normal state of brain functional organization. 

Reorganization of surviving central nervous system elements supports behavioral 

recovery. In this framework, the development of therapeutic approaches whose goal is to 

promote repair and restoration of function after stroke could be strategic. Two of the 

most prominent molecular events occurring in brain ischemia are inflammation and 

overactivation of the NMDA subtype of glutamate receptors (excitotoxicity), which lead to 

the progression of neuronal damage. Although this, an endogenous neuroprotective 

response, at least partially mediated via induction of neuroprotective factors, is also 

elicited by ischemic injury. Actually, the great potential of the brain for reorganization, 

plasticity and repair after injury is a critical issue to be taken into consideration, that has 

previously been understimated. 

Knowing the pathophysiological mechanisms that follow ischemia and their time course is 

essential in order to be able to identify and use biomarkers in monitoring the patients 

with acute stroke. Although still in the research phase, some biomarkers have been 

shown to correlate with the clinical evolution and could be important in foreseeing the 

clinical response to the treatment of the acute phase and to the different rehabilitation 

strategies in the post-acute and chronic phases. In addition, the inflammatory and 

neuroprotective response during the evolution of stroke can be monitored in the patient 

through suitable techniques such as neuroimaging. 

Up to now research on molecular targets in cerebral ischemia for developing novel 

therapeutics has mainly focused on single pathways, being inflammation or excitotoxicity 

or oxidative stress. The step forward, to go beyond the current state of the art, of the 

present project is to try to consider a group of different concurrent factors elicited during 

stroke. The project aims at investigating the cross-talk between inflammation, 

excitotoxicity, and endogenous neuroprotective/neuroregenerative pathways. In 

collaboration with the Neurological Institute C. Mondino, levels of selected biomarkers 

representative of inflammation and neuroprotection will be monitored in the serum of 

patients with stroke at different time points and then will be correlated with 

neuroimaging data and the clinical outcome. This will

provide a clinical basis to finalize the basic research focussed on the 

evaluation of the molecular mechanisms recruited in the cross-talk 

between inflammation, excitotoxicity, and endogenous 

neuroprotective/neuroregenerative pathways. To this purpose we will 

evaluate in glial and neuronal population (i) the impact of inflammation 

on expression of neuroprotective factors relevant to brain ischemia (i) 

the effect of neuroprotective factors on inflammation, (ii) the effect of 

cytokines and neuroprotective factors on excitotoxicity driven by the 

NMDAR 

The project is designed to identfy new prognostic markers and new 

targets for therapy, aimed at boosting or superinducing the endogenous 

neuroprotective response directly or interfering with the mechanisms 

that blunt this response in conditions of injury.

Nome ANGELINI - ACRAF SpA 

Contatti 

Tel. 

E mail 

Claudio Milanese 

06-91045290 

c.milanese@angelini.it 

Istituto/Dipartimento Angelini Research Center- S.Palomba (Rome), Italy 


Area di interesse identificata Study of the role of pro-inflammatory chemokines in 

neuroinflammatory disorders and analysis of the therapeutic potential 

of chemokine synthesis inhibitors 

Abstract 

Chemokines and their receptors play a crucial role in the trafficking 

of leukocytes and other immune system cells and are of particular 

interest in the context of the unique immune responses elicited in 

the central nervous system (CNS). Within this particular district, 

most data focus on the role of CCL2 and the related factors CCL7 

and CCL8, since they have been implicated in a wide range of 

neuropathologies, including trauma, ischemic injury and multiple 

sclerosis (Semple BD et al., 2010, J Cerebral Flow & Metabolism 

30:459). Furthermore, members of this chemokine family and their 

receptors recently emerged as key modulators in nociceptive influx 

transmission in neuropathic and inflammatory chronic pain models 

(Gosselin RD et al., 2008, Curr Med Chem 15:2866), providing 

evidence of the involvement of chemokines as important mediators 

of the initiation and maintenance of pain hypersensitivity. 

Angelini has a large experience in the study of chemokine synthesis 

inhibitors. 

A small original Angelini molecule able to inhibit the synthesis of 

CCL2, bindarit, has recently successfully concluded a phase II clinical 

study in diabetic nephropathy and it is currently undergoing 

another phase II study in restenosis. The aim of the present project 

is the identification and development of new molecules able to 

inhibit the synthesis of chemokines and endowed with physicochemical 

and ADME characteristics useful to target CNS disorders. 

The evaluation of the pharmacological activity of such compounds 

in appropriate models will allow to assess their potential as 

therapeutic agents useful for the treatment of CNS disorders and 

chronic pain. 






ISTITUTO SUPERIORE DI SANITA’ (Referente, Luisa Minghetti) 

The Istituto Superiore di Sanità is the leading technical and scientific body of the Italian National Health 

Service. Its activities include research, clinical trials, control and training in public health. The Institute 

conducts scientific research in a wide variety of fields and also serves as a major source of information 

relating to public health and biomedicine in Italy through online connections to national and international 

scientific databases and data banks. 

The institute has been actively involved in research in ageing and related neurodegenerative diseases over 

the last decade. It is partner in the European Research Area in Ageing network (ERA-AGE and ERA-AGE2) 

funded under the Sixth and the Seventh Framework Programme. 

The research on Alzheimer’s disease in ISS takes advantage of multidisciplinary expertise and resources 

including proteomics, molecular and cellular biology, neuroimmunology, behavioral analysis, in vitro and in 

vivo models, genetic, clinical and epidemiological studies, advanced statistical analysis, data and bio-banks. 

Main topics of research are: in vitro toxicity of amyloid proteins; excitotoxicity and free radical-related 

alterations of signaling pathways and post-translational modifications of post-synaptic machinery; analysis 

of protein aggregate characteristics responsible for the induction of amyloid formation; characterization of 

relevant rodent models by pre-clinical imaging with MRI scanning; characterization of behavioral and 

cognitive alterations in relation to neurochemical/neuroanatomical markers in AD murine models; effects 

of diet, pharmacological agents, or neurotoxicants on disease susceptibility and progression in AD murine 

models; early environmental factors in modulating onset and progression of neurodegenerative diseases; 

self antigens and autoantibodies in the pathogenesis of AD and as biomarkers of the disease; peripheral 

oxidative stress and antioxidant defense related biomarkers as prognostic and diagnostic tools; 

identification of mutations and polymorphisms affecting clinical phenotype and progression; descriptive, 

analytical, clinical epidemiological studies; longitudinal data bases for cohorts of elderly persons with 

cognitive deficits (IPREA and ILSA). 

Research projects currently active include: 

• Role of Genetic Determinants on the Progression of Clinical Phenotype from MCI to Alzheimer’s Disease. 

PI: A.Confaloni (National coordinator F.Tagliavini), Italian Ministry of Health .Eur 60,000 

• Determinanti genetici e fattori modulatori nelle malattie neurodegenerative: modelli clinici ed animali - 

P.I. A. Confaloni (National coordinator A. Bruni), Italian Ministry of Health, Eur 70,000 

• Risk Assessment of iatrogenic transmission of neuro-AMYloidoses (RAAMY). PI: M.Pocchiari (in 

collaboration with E.Comoy, CEA, France), Alliance BioSecure, Fondation de recherche reconnue d’utilité 

publique. 

• Early diagnosis of Alzheimer’s diseases through the combination and correlation of molecular, functional 

and clinical biomarkers PI: M.Pocchiari, Italian Ministry of Health. Eur. 364,000 

• Innovative approaches to the study of induction/repair of DNA oxidative damage in models of 

neurodegenerative diseases: molecular basis and identification of possible therapeutic targets. PI: 

P.Popoli, Italian Ministry of Health. Eur 364,000 

• Effects Of Dog-Assisted Therapies On Physical And Psychological Well Being In The Institutionalized 

Elderly. PI: F.Cirulli, ISS and Nando Peretti Foundation, Eur 100,000 

• Therapeutic effects of human-animal interactions in a rural environment on neuropsychiatric diseases 

and brain aging. PI: F.Cirulli, INEA Eur 62,000 

• Nuovi farmaci ad azione neurotrofica e neuroprotettiva per applicazioni terapeutiche nelle neuroscienze: 

malattia di Azheimer, malattia di Huntington e Sclerosi Laterale Amiotrofica. PI: D. Merlo, (National 

coordinator A. Cattaneo). MIUR – FIRB “Idee Progettuali RBIP063ANC” 2007, Eur 32,000

• Ruolo neuroprotettivo della Timosina beta4 in modelli sperimentali in vitro di eccitotossicità, ischemia e 

Alzheimer Disease”. PI: D. Merlo, Tecnogen Spa, Eur 125,000 

• Role of recurrent herpetic infections in neuronal damage. PI D. Merlo (National coordinator 

A.T.Palamara), Ministero della Salute, Eur 70,000 

• An integrated approach to identify functional, biochemical and genetic markers for diagnostic and 

prognostic purposes in the elderly, in the centenarians and in people with dementia, Alzheimer’s disease, 

mild cognitive impairment. PI: N.Vanacore, Italian Ministry of Health, Eur 291,000 

• Alzheimer's Disease (AD) and antipsychotics: a long term, multicentre, double blind, randomised clinical 

trial. PI: R.Raschetti, Italian Agency of Drug, Eur 1,950,000. 

• EC 7th Framework Programme “ERA AGE 2 Project – European Reasearch Area in Ageing Extension” . 

E. Scafato. 

EC 7th Framework Programme “ FUTURAGE Project – A Roadmap for Ageing Research” . E. Scafato 

• EC 2nd Programme of Community Action in the Field of Health “VINTAGE Project – Good Health into 

Older Age”. E. Scafato. 

• EC 5th Framework Programme “DESCRIPA Project - Development of screening guidelines and criteria for 

predementia Alzheimer’s disease”. E. Scafato.

Nome Paola Bossù 

Contatti 

Tel. 

E mail 

Via Ardeatina 306, I-00179, Rome 

06-51501520 

p.bossu@hsantalucia.it 

Istituto/Dipartimento IRCCS Fondazione Santa Lucia/Clinical and Behavioral Neurology 


Role of inflammation in Alzheimer’s Disease: study of pro-inflammatory cytokines for the 

identification of new pathogenic and diagnostic targets 


Inflammation and neurodegeneration 

identificata 






Genetic Risk factors and peripheral biological markers of conversion 

from Mild Cognitive Impairment to Alzheimer’s Disease 

Italian Ministry of Health 

01/2009 - 01/2011 

Several studies have demonstrated that inflammation plays an 

important role in mediating glial alterations and neurodegeneration 

that occur in AD patients. This phenomenon includes the accumulation 

of reactive microglia and astrocytes in damaged regions of AD brain 

and increased pro-inflammatory cytokine expression that may further 

contribute to the development and progression of pathological state. 

Accordingly, in several studies pro-inflammatory cytokines such as 

Interleukin (IL)-1, IL-6, Tumor Necrosis Factor (TNF)-α and IL-18 

have been found to be peripherally increased in AD patients and 

substantially impaired in the late stage of the disease. However, the 

temporal definition of their implication in respect to disease 

progression, as well the extent to which inflammatory factors 

participate to the neuronal damage are issues of fundamental 

importance, which are to date still unclear. Recent studies performed 

by this participant unit have identified a possible role for the proinflammatory 

cytokine IL-18 in AD, by means of genetic data 

suggesting that IL-18 gene promoter polymorphisms can predict risk 

and outcome of AD, and by demonstrating that IL-18 is overexpressed 

by blood cells of AD patients and correlates with their cognitive 

decline. In the present study, by using a longitudinal clinical approach, 

the previous results regarding IL-18 will be expanded also to other 

inflammatory mediators with potential role in the disease progression 

and pathogenesis. In fact, this project is aimed to investigate the 

impact of pro-inflammatory and anti-inflammatory parameters during 

disease progression in patients at the higher risk of developing AD, i.e. 

MCI patients. These objectives are relevant for the identification of 

new diagnostic markers of the AD preclinical form and for the 

comprehension of inflammatory mechanisms, likely participating in 

the earliest pathogenic processes of AD.


D. Oxidative Stress and neurodegeneration

Nome Elisa Greggio, Marco Bisaglia, Luigi Bubacco 

Contatti 

Via Ugo Bassi 58/b, 35129 Padova, ITALY 

Tel. +39 049 8276244 

Istituto/Dipartimento Dipartimento di Biologia, Università di Padova 


Area di interesse identificata Indagine dei meccanismi molecolari alla base della malattia di Parkinson 



Mitochondrial dysfunction and axonal transport in genetic models of 

Parkinson’s disease 

Ente finanziatore MIUR 


Abstract del progetto Mitochondrial dysfunction and oxidative stress have been implicated 

in the pathogenesis of Parkinson’s disease (PD). Defective axonal 

transport, including abnormal accumulation of misfolded proteins and 

organelles and damage of transport of cargoes through axons, is also 

observed in many neurodegenerative diseases. Few lines of evidences 

suggest that both alpha-synuclein and LRRK2, proteins involved in 

dominantly transmitted forms of familial PD, may play a role in 

mitochondrial function, microtubulo stability/dynamics and axonal 

transport. The aim of this project is to use cellular and, in part, animal 

models of !-synuclein and LRRK2 to explore the 

physiological and pathological role of this two genes in mitochondrial 

functionality and in the transport of mitochondria and synaptic vesicle 

along the axons. 


Biochemical characterization of full length human recombinant LRRK1 

and LRRK2 

Ente finanziatore Michael J Fox Foundation 

Durata progetto 1+1 years 

Abstract del progetto Mutations in one gene, LRRK2, are a common cause of Parkinson’s 

disease. However, the protein product of this gene is large and complex 

and, to date, no one has been able to make highly pure, functional LRRK2 

that can be easily used for biochemical experiments. The primary 

objective of this project is to make purified protein by combining 

expertise in multiple technologies from each of our collaborating 

laboratories and to share our methods with other researchers. If 

successful, our secondary objective is then to perform an exhaustive 

search for other proteins that may interact with LRRK2. 


Molecular mechanisms that involve the oxidation product of dopamine 

in the pathogenesis of Parkinson disease. 

Ente finanziatore Prin- MIUR 

Durata progetto 1+1 years 


Several hypotheses have been proposed in the literature to describe 

the mechanisms that lead to neurodegeneration in PD. This project will 

focus on the ossidative stress associated to the redox chemistry of 

dopamine. The rational in this choice is that dopamine oxidation may 

reasonably account for the specificity observed for dopaminergic neuron

degeneration in PD. Although the oxidation of DA and DOPA lead to the 

formation of toxic ROS, the main cytotoxic species which could potentially 

explain the specific damage to dopaminergic neurons remain the highly 

reactive quinone species (DAQ) that have many potential protein targets 

for chemical modifications. The identification of several proteins, whose 

functions are affected by the mutations associated to familiar PD, 

triggered the hypothesis that late onset idiopathic forms of PD may 

derive from similar functional effects induced by chemical modifications 

produced by DAQ. This project will be focus on the definition of the 

damages induced by DAQ on two cytoplasmatic (DJ-1 and α-Synuclein) 

and two mitochondrial (PINKI and Superoxide Dismutase 2) possible 

protein targets.

Nome Antonella Bobba 

Contatti Tel: 080 5442412 Fax: 0805443317 E-mail: a.bobba@ibbe.cnr.it 

Istituto/Dipartimento Istituto di Biomembrane e Bioenergetica (IBBE), CNR 

Via Amendola 165/A 70126 Bari, Italy 



identificata 



MITOCHONDRIA, APOPTOSIS AND NEURODEGENERATION: CHARACTERIZATION OF 

MITOCHONDRIAL DYSFUNCTION IN NEURODEGENERATIVE DISEASES 

There is compelling evidence for the direct involvement of mitochondria in 

neurodegenerative disorders. In most cases, increased oxidative stress appears to 

be the crucial initiating event that affects respiratory chain function and starts a 

vicious cycle finally leading to neuronal cell death via apoptosis or necrosis. In 

addition a functional interaction exists in the cells between cytoplasm and 

mitochondria and the cross-talk ability of these cellular components is involved in 

cell homeostasis and death. In the light of this, the goal of this topic is to ascertain 

whether and how, during neurodegeneration, the energy metabolism in the 

cytoplasm changes as a result of mitochondrial alterations and vice-versa and the 

mechanism by which certain compounds can prevent cell death. 

CHARACTERIZATION OF THE MOLECULAR MECHANISMS UNDERLYING AGING, CELL SENESCENCE 

AND NEURODEGENERATIVE DISEASES 

Aberrant modulation of cell signaling pathways involved in cell death regulation 

underlies several human pathological conditions, such as neurodegeneration. 

Although the mechanistic aspects of cell death and growth have been extensively 

investigated, there is still lack of knowledge about the early events that lead to 

deregulation of cell homeostasis and about the mechanisms of cell sensitization to 

growth and death signals in response to changes in cell environment. Thus, it is 

worthwhile to characterize the progression of the biochemical events and in 

particular of those occurring in the early phase of neurodegeneration with the aim 

to identify the critical steps that once blocked could allow for the restore, even if 

partial, of the impaired cellular function/activity. The characterization of the main 

signalling pathways that are activated and/or impaired during neurodegeneration 

and the close definition of the cause-effect relationship between them are powerful 

tools which are potentially usable to design new diagnostic, prognostic and 

therapeutic approaches. To this aim it is of great relevance an approach in which 

functional genomics and whole-genome expression profiling are combined in order 

to efficiently identify key players in biological response pathways. This is possible if 

different cellular systems are complementarily used in the experimental approach. 

Neurotrofine e meccanismi relativi a malattie neurodegenerative. 

UO3 - Stress ossidativo e bioenergetica mitocondriale nella patogenesi delle 

malattie neurodegenerative 

Ente finanziatore MIUR (Progetto CNR/MIUR - Fondo FISR) 

Durata progetto 2 years (11/06/2003 – 1/10/2005) 

Abstract del progetto Studies on the involvement of mitochondria and role of oxidative stress in the 

ethiopathogenesis of neurodegenerative diseases by using primary neuronal cell 

cultures as in vitro model. The main results can be summarized as follow: i) early 

impairment of the energetic metabolism; ii) released cytochrome c exerts a crucial 

role by acting both as a ROS scavenger and an electron donor other than a caspase 

activator; iii) ATP level increases in the early phase of the death process and iv) ROS 

and thereafter activated caspases induce the progressive impairment of the

mitochondrial ANT. 


Malattie neurodegenerative come conseguenza di un alterato processamento di 

proteine neuronali, modelli animali e di colture cellulari in vitro 

Ente finanziatore MIUR (Progetto FIRB cod. RBNE01ZK8F) 

Durata progetto 3 years (08/01/2003 – 10/01/2007) 

Abstract del progetto Both animal and in vitro cultured cellular models have been used to study the 

molecular mechanisms which are at the basis of some neurodegenerative diseases 

and to identify new, potential therapeutical tools. Studies have been mainly focused 

on a thorough analysis of the mitochondrial function impairment with collapse of 

oxidative phosphorylation and increased production of reactive oxygen species. 


Bioenergetic and apoptotic systems of mitochondria. Genomics, proteomics, cellular 

homeostasis and physiopathology. 

WP2- Mitochondria role in animal, plant and microorganism cell apoptosis 

Ente finanziatore MIUR (Progetto MIUR N. 157) 

Durata progetto 3 years (2006-2009) 

Abstract del progetto The aims of the research project are concerning with: a)The role of mitochondria in 

the progression of neuronal apoptosis; b)The role of mitochondria in the apoptosis 

of a plant model system; c)The role of mitochondria in yeast apoptosis; d)The role of 

mitochondria in the induction/resistance to apoptosis in neoplastic cells 

Titolo progetto Molecular bases in ageing-related degenerative syndromes 

Ente finanziatore MIUR - Progetto MERIT - Protocollo: RBNE08HWLZ 

(UR n 11: Protocollo: RBNE08HWLZ_012) 

Durata progetto 3 years (approved but not yet funded) 

Abstract del progetto The aim of this project is to discover specific molecular pathways and/or targets for 

the developing of new therapeutic strategies. Research activities are grouped into 

four workpackages: 1) Aggregation states and cytotoxic activity: molecular and in 

vivo/in vitro biological studies. 2) Oxidative stress and ageing: biochemical 

processes and regulation of gene expression. 3) Dismetabolic factors and 

neurodegeneration: molecular, biophysical and biochemical studies. 4)Exogenous 

and endogenous molecules against ageing and neurodegeneration: application for 

the diagnosis and therapy. 


Biotechnology applied to the study and utilization of Apulian extra-virgin olive oil 

and its minor components in aging and in diseases associated to oxidative stress 

Ente finanziatore Regione Puglia (Progetto Reti di Laboratorio Pubblici di Ricerca) 

Durata progetto 3 years (approved but not yet funded) 

Abstract del progetto Constitution of a Laboratories Network aimed to identify the nature and efficacy of 

minor components of Apulian extra virgin olive oil in the prevention of oxidative 

stress-associated pathologies

Nome Lenaz Giorgio and Giancarlo Solaini 

Contatti 

051-2091229; 333-2383218; fax: 051-2091217 

giorgio.lenaz@unibo.it 

051-2091215 ; 339 1948613 ; fax 051-2091224 

giancarlo.solaini@unibo.it 

Istituto/Dipartimento Dipartimento di Biochimica 


The involvement of mitochondria in neurodegenerative diseases, including Alzheimer disease (AD) is 

widely documented, although no clear causal relationships are available for a great number of 

scattered observations. Both a decrease of mitochondrial respiration, in particular of cytochrome 

oxidase and an enhanced production of Reactive Oxygen Species (ROS) has been reported in autoptic 

samples of AD patients and in neuronal cells of animal models of AD. Most researchers in the field 

believe these findings being the consequence of toxic amyloid peptides (A-beta) interacting with 

mitochondrial structures, and several studies have recently shown that A-beta and tau proteins, 

characteristic of AD, interact with mitochondrial respiratory complexes, opening novel hints to 

understand the pathogenesis of the disease. A significant investigation on the above issue has been 

carried out by some of us, but it’s noteworthy that our research group has a world wide reputation 

on the study of mitochondrial structure and function and has already been involved in research on 

aging and neurodegenerative diseases including AD. In particular we have shown the functional 

consequences of the recently discovered super-association of respiratory complexes and 

demonstrated that these super-complexes are easily dissociated by a number of changes in proteinprotein 

and lipid-protein interactions. We intend to propose and test the following working 

hypothesis. Under conditions likely present in brain regions of AD patients, the interaction of A-beta 

and tau with mitochondria at the level of respiratory chain is able to disrupt super-complex 

associations with consequent destabilization of the individual complexes, in particular Complex I 

(NADH Coenzyme Q reductase). This would necessarily determine an enhancement of production of 

ROS, thus explaining the observed oxidative stress in AD mitochondria. Loss of efficient electron 

transfer and altered assembly of Complex I would determine decreased oxidative phosphorylation 

and at the same time the oxidative stress could represent a sufficient stimulus for triggering cell 

death by necrosis or apoptosis. Our laboratory has the expertise and technology for all investigations 

required to test this hypothesis, and we are currently setting conditions considered likely to occur in 

brain regions of AD patients. The study will be performed on animal model systems and in human 

cells cultures. On the other hand, a better understanding of AD pathogenesis might establish the 

basis for attempts of therapeutic interventions, including metabolic therapy as recently shown in our 

laboratory for diseases featuring alterations of ATP synthesis. 

Area di interesse identificata Basic research/new treatment strategies 

Finanziamenti ricevuti


Ente finanziatore MIUR (PRIN 2008) 



Ruolo dell’organizzazione sopramolecolare della fosforilazione 

ossidativa mitocondriale nella produzione di specie reattive 

dell’ossigeno e in alterazioni patologiche in sistemi modello e in 

cellule umane. 

In this project we will test the working hypothesis that an initial 

enhanced ROS generation due to different possible reasons and 

originating in different districts of the cell besides mitochondria 

would induce disorganization of the supercomplex assemblies of the 

oxidative phosphorylation system, eventually leading to instability 

of Complex I quaternary structure; both the lack of efficient electron 

channelling and the decrease of Complex I would cause a fall of 

NAD-linked respiration and ATP synthesis, leading to pathological 

changes. This hypothesis will then be tested in two pathologies in 

which a major pathogenic factor is oxidative stress, i.e. the NARP 

syndrome (neuropathy with ataxia and retinitis pigmentosa) due to 

mitochondrial DNA mutations in the ATP6 gene, and diabetic 

peripheral neuropathy, a common and severe complication of 

diabetes mellitus. (Details follow)

Nome Annalisa Santucci 

Contatti 

Tel. +390577234958 

Fax +390577234903 

Email santucci@unisi.it 

Istituto/Dipartimento Department of Molecular Biology, University of Siena, ITALY 


Amyloid β peptide (Aβ) is directly involved in the pathogenesis of Alzheimer Disease (AD) or tightly 

correlated with other primary pathogenic factors. The predominant forms of Aβ in the human brain are 

Aβ(1-40) and Aβ(1-42), but Aβ(25-35) fragment, physiologically present in elderly people [1], is the more 

toxic region and has been recently found to play a relevant role in AD, due to its peculiar aggregation 

properties. In fact, actually, the ability to affect cognitive processes is typical not only of the full-length 

peptide, but also of several Aβ fragments, in particular just the undecapeptide Aβ(25-35). This peptide is 

regarded as the functional domain of Aβ, responsible for its neurotoxic properties and represents the 

actual biologically active region of Aβ. In vivo, Aβ(25-35) is present in neurons of subiculum and entorhinal 

cortex of AD brains, and it was also observed in Inclusion-Body Myositis (IBM) muscle. Self-assembly of 

Aβ(25-35) is a key player in AD, since it could nucleate the assembly of Aβ monomers and a bent in this 

region could be the rate-limiting step in Aβ fibril formation. In virtue of this, there is increased interest in 

the role of Aβ(25-35) fragment in free radical-associated neurotoxicity in AD brain. Age-dependent 

racemisation of Ser residue at the 26th position in Aβ(1-40) is critical in AD pathogenesis. In vivo 

conversion of non-toxic [DSer26] Aβ(1-40) to toxic [D-Ser26]Aβ(25-35) has been finally demonstrated and 

the presence of truncated and toxic [DSer26] Aβ(25-35) in AD brains, but not in age-matched control 

brains, adds a major value to in vivo neurodegeneration mechanism. Given the suggested centrality of 

Aβ(25-35) to the pathogenesis of AD, this peptide should deserve major attention also as a therapeutic 

target. In fact, diagnosis and monitoring of sporadic AD have long depended on clinical examination of 

individuals with end-stage disease. However, upcoming anti-AD therapies are optimally initiated when 

individuals show very mild signs of neurodegeneration. Our research group provided for the first time an 

exhaustive overview about Aβ(25-35) [2] and during the last few years it has deepened the aggregation 

properties of this peptide according to solution and environmental conditions [3]. Actually, we are 

dedicating attention to this peculiar peptide in the light of its great oxidative stress generation capacity 

and extreme toxicity in neuronal cells and synaptosomes. The evidence points toward Aβ(25-35) being 

primarily responsible for neurodegeneration observed in AD. The question remains on how this can induce 

such degenerative effects. Strong of our experience in the field of the study of oxidative stress [4-9], we 

propose ourselves to estimate the mechanisms been involved in the ROS generation Aβ(25-35)-mediated 

in different cell models. We can use SELDI technology to study [DSer26] Aβ(25-35) as a core biomarker for 

the mild cognitive impairment stage of AD. From a longer perspective, establishment of the most effective 

combinations of different biomarkers and other diagnostic modalities may be foreseen. Moreover, due to 

our well established experience in Proteomics [10-15], we will apply proteomics techniques to study cellspecific 

early markers of brain aging-related degeneration in human astrocytes and neurons that may 

contribute to neurodegenerative damage. The protein expression patterns of the AD neuron and astrocyte 

cultures will be compared with those obtained from healthy people. Differentially expressed spots will be 

identified by matrix-assisted laser desorption/ionization-time of flight peptide map fingerprinting and 

database search. A number of the protein alterations have been previously reported in the brain tissue 

proteome of animal models, aged brain or AD brain, but are still quite unexplored in neuron and astrocyte 

proteome and our project could help to clarify if both cell types are involved in the brain degenerative 

changes as well as to study neuronal changes having a greater influence in the pathology progress. These 

findings will be discussed in reference to the effect of specific protein oxidation and changes of expression

on potential mechanisms of abnormal alterations in metabolism and neurochemicals, as well as to the 

learning and memory deficits in AD. 

References 

[1] Kubo, T.; et al. A possible mechanism of neurodegeneration in AD - Involvement of racemized βamyloid. 

Soc. Neurosci., 1999, Abstr. 25, p. 838. 

[2] Millucci L, et al. Conformations and biological activities of Amyloid Beta Peptide 25-35. Curr Protein 

Pept Sci. 2009 Sep 15. 

[3] Millucci, L., et al. Rapid aggregation and assembly in aqueous solution of Aβ(25-35) peptide. J. Biosci. 

2009, 34(2), 293-303. 

[4] Braconi D, et al. Proteomics and redox-proteomics of the effects of herbicides on a wild-type wine 

Saccharomyces cerevisiae strain. J Proteome Res. 2009;8(1):256-67. 

[5] Braconi D, Possenti S, Laschi M, Geminiani M, Lusini P, Bernardini G, Santucci A. Oxidative damage 

mediated by herbicides on yeast cells. J Agric Food Chem. 2008 May 28;56(10):3836-45. 

[6] Braconi D, et al. Comparative analysis of the effects of locally used herbicides and their active 

ingredients on a wild-type wine Saccharomyces cerevisiae strain. J Agric Food Chem. 2006 19;54(8):3163- 

72 

[7] Desideri G, et al. Effects of bezafibrate and simvastatin on endothelial activation and lipid peroxidation 

in hypercholesterolemia: evidence of different vascular protection by different lipid-lowering treatments. J 

Clin Endocrinol Metab. 2003;88(11):5341-7. 

[8] Desideri G, et al. Angiotensin II inhibits endothelial cell motility through an AT1-dependent oxidantsensitive 

decrement of nitric oxide availability. Arterioscler Thromb Vasc Biol. 2003;23(7):1218-23. 

[9] Trabalzini L, et al. Proteomic response to physiological fermentation stresses in a wild-type wine strain 

of Saccharomyces cerevisiae. Biochem J. 2003;370(Pt 1):35-46. 

[10] Bernardini G, et al. Postgenomics of Neisseria meningitidis for vaccines development. Expert Rev 

Proteomics. 2007;4:667-77. 

[11] Bernardini G, et al. The analysis of Neisseria meningitidis proteomes: Reference maps and their 

applications. Proteomics. 2007;7(16):2933-46. 

[12] Spreafico A, et al. A proteomic study on human osteoblastic cells proliferation and differentiation. 

Proteomics. 2006;6(12):3520-32. 

[13] Mini R, et al. Comparative proteomics and immunoproteomics of Helicobacter pylori related to 

different gastric pathologies. J Chromatogr B Analyt Technol Biomed Life Sci. 2006;833(1):63-79. 

[14] Bernardini G, et al. Proteome analysis of Neisseria meningitidis serogroup A. Proteomics. 

2004;4(10):2893-926. 

[15] Magnani A, et al. Two-step elution of human serum proteins from different glass-modified bioactive 

surfaces: a comparative proteomic analysis of adsorption patterns. Electrophoresis. 2004;25(14):2413-24. 

Area di interesse identificata Post-Genomic Biochemistry of Neurodegenerative Diseases 


Titolo progetto Characterization of peptide oligomers in Alzheimer Disease 

Ente finanziatore Siena Biotech s.p.a. 

Durata progetto 1/07/2006 - 30-04-2009 

Abstract del progetto Aβ(25-35) is the shortest peptide sequence that retains biological activity 

of full-length Aβ(1-42) and exhibits large β-sheet aggregated structures. 

Because of these features, it has often been chosen as a model for fulllength 

Aβ in structural and functional studies but kinetic and structural 

studies of this peptide are indeed strongly hampered by its tendency to 

quickly assemble into insoluble aggregates. The aim of the project was to 

study the uncommon rapidity with which Aβ(25-35) self-assembles at 

different pH. The obtained results support the hypothesis that the early 

Aβ(25-35) aggregated forms, that develop at pH 3, may not enhance the 

growth of dangerous fibrils generated at pH 7.4. We conclude that the

mechanism by which Aβ forms toxic fibrils is initiated in vivo and probably 

does not involve low pH compartments, confirming the hypothesis of Kubo 

et al. (2002) on the origin and toxicity of Aβ(25-35).


E. Trophic factors involvement in the 

neurodegeneration process


F. Endocrine and Metabolic Factors

Country: Italy 

Contact person: Paola Fragapane, Cecilia Mannironi (CNR) 

Date: 28/01/2010 

I) Strategic Issues 

Stress is a term to describe experiences that are challenging both emotionally and physiologically. 

A feature of the stress response is the activation of the autonomic nervous system and 

hypotalamus-pituitary-adrenal (HPA) axis with the production of glucocorticoids hormones. This 

response is needed to survive dangerous situation but excessive adrenocortical and autonomic 

function is deleterious for health and survival. The central nervous system in response to such 

activation elaborates the appropriate behavioral and physiological responses. As regards to the 

neuroanatomical substrate the hypothalamus and the brain stem are essential in the response to 

stressors, however, other brain regions are also well known targets of stress hormones. Among 

such regions, research has focused its attention on the prefrontal cortex, the hippocampus and the 

amygdala. Interestingly these brain structures have been related also to functions such as 

memory, and decision making but also to anxiety and other psychiatric diseases. 

Acute (single stress) and chronic (repeated) stress induce prominent changes in neuronal activity 

and synaptic functions. These changes are thought to be due to neuronal remodelling such as 

dendrites shortening and debranching (Mc Ewen, 2007) in the hippocampus and in the medial 

prefrontal cortex. These stress-induced changes are mediated by modification of gene expression 

at transcriptional levels. However, recent studies are pointing out the contribution of 

posttranscriptional regulation to stress associated responses (De Rijk et al., 2003). An important 

post-transcriptional mechanism of gene regulation involves microRNA (miR). A range of evidence 

suggests that these small RNAs form complex networks involved in the regulation of differentiation 

and development, via regulation of chromatin modification, transcription, translation and RNA 

stability. They are small RNA molecules (about 22 nucleotide long) that inhibit mRNA translation or 

induce mRNA degradation by sequence specific pairing with mRNA 3' untranslated regions 

(UTRs). In the vertebrate nervous system miR have been shown to play a role during development 

and neurogenesis (Kosik, 2009; Lagos-Quintana et al., 2001). MiR are also abundantly expressed 

in the adult brain and appear to regulate the maintenance of mature trait and synaptic plasticity 

(Mehler and Mattick, 2006). MiR134 was reported to be a regulator of dendritic spine development 

in brain (Schratt et al., 2006). Perturbation in miR are associated with a number of neuronal 

diseases as X-linked mental retardation, Parkinson’s disease and schizophrenia. 

II) Priority areas 

The molecular mechanism underlying brain response to stress are still incompletely understood. 

The aim of this project is to identify region-specific stress-induced changes in miR expression. The 

expression profiles of miRs in brain regions known to be involved in stress response such as the 

hippocampus, the amygdala and the prefrontal cortex will be analyzed after the exposure to 

aversive stimuli. The analysis will be performed in rodents exposed to single or repeated restraint

stress and gene expression in each brain regions will be compared with that of matched control 

mice. LNA (locked nucleic acid) microarrays will generate a limited set of candidate miRNAs, which 

will be validated by Northern blots and qPCR. In order to have a better neuroanatomical definition 

of miR expression selected miR will be analyzed by in situ hybridization. mRNA targets it will be 

predicted through the use of freely available algorithms and will be validated by reporter construct 

expression in cell system. Moreover bioinformatic analysis of miR upstream promoter sequences 

will allow us to identify transcription factors involved in their modulation in response to stress 

stimuli. 

III) Impact 

The hippocampus, the amygdala and the prefrontal cortex are interconnected and influence each 

others via direct and indirect neuronal activity. It is well established their involvement in the stress 

response and in the plastic changes occurring in response to acute and repeated stress exposure. 

Our analysis of miR expression pattern will allow us to understand possible region specific 

expression pattern and/or common molecular networks. 

Translation of the information on stress effects from animal models to the human organism is an 

important challenge. Indeed maladaptive response to stress is considered causal to many 

psychiatric diseases such as major depression, anxiety as well as schizophrenia. The 

understanding of the contribution of non coding RNAs might allow on the one hand a better 

understanding of the neural mechanisms at the basis of stress response on the other hand might 

help identify possible targets for the development of new future therapies.

Nome Roberto Rimondini-Giorgini 

Contatti roberto.rimondini@unibo.it 

Istituto/Dipartimento Department of Pharmacology 


Via Irnerio 48 

40126 Bologna 

Italy 

Area di interesse identificata Developing competitive animal models / basic research 



MECHANISMS BEHIND THE ASSOCIATION OF APOE4 GENOTYPE 

AND LIFE STYLE RELATED FACTORS IN ALZHEIMER`S DISEASE 

Ente finanziatore Fondazione Cassa di Risparmio di Bologna 



Alzheimer`s disease (AD) is a devastating neurodegenerative disease 

and the most common cause of dementia. The current estimate 

number of people with dementia in the world stands at 24.3 million, 

with an estimated 4.6 million new cases every year in the world. 

There are no definitive diagnostic tests, biological markers of AD or 

pharmaceutical treatments. At central nervous system (CNS) level, 

the pathological signs include deposits of extracellular amyloid-β 

peptide (Aβ) in plaques. Loss of neurons and synapses is also 

widespread. Whilst some cases have a genetic component (familiar 

AD) the majority of cases (90-95%) have unknown cause and are 

named sporadic AD. Familiar AD seems to be correlated to 

mutations in key genes such as in the amyloid-precursor-protein 

(APP) gene and in the presenilin genes (1 and 2). On the contrary, 

sporadic AD seems to be consequence of a complex interaction 

between both genetic and environmental risk factors. The major 

genetic risk factor is an allelic variant of apolipoprotein E (apoE) 

called apoE4. Nowadays, no specific environmental risk factor has 

been definitively identified as being associated with AD. Anyway, AD 

is associated with a history of depressive illness, traumatic head 

injury, cardiovascular disease, smoking and stroke. In addition, 

several evidences highlight the correlation between high alcohol 

consumption, saturated fats and cholesterol intake. The 

apolipopoteins are cholesterol transporters of high importance for 

neuronal plasticity, CNS glucose utilization and mitochondrial

functions. The apoE isoforms (2, 3 and 4) differ in their abilities to 

accomplish these critical tasks. Approximately 20% of the 

population carries one or two apoE4 alleles. ApoE4’s involvement in 

neuropathology is well documented factor for AD in many 

populations. In some, 40–80% of patients with AD possess at least 

one apoE4 allele. While ApoE3 and apoE2 are effective in 

maintaining brain homeostasis, apoE4 seems to have an opposite 

rule. Impaired cognition in non-dement individuals carrying the 

apoE4 allele worsens with age, suggesting a global detrimental 

effect on the CNS. Although apoE4 is strongly linked to AD 

pathology, its mode of action is unknown. Insights into the role of 

apoE in neuropathology have come from studies of transgenic mice 

expressing human apoE3 or apoE4 in neurons or astrocytes. 

Features of AD pathology in these animals include reduced numbers 

of presynaptic terminals in mice expressing apoE4 with or without 

expression of human amyloid precursor protein (APP), increased 

plaque deposition in apoE4 mice expressing APP, increased 

phosphorylation of tau, impaired learning and memory, and altered 

long-term potentiation. No studies are available on the correlation 

between life style and apoE4 genotype in AD incidence. Aim of this 

project is to study the impact of different life conditions in relation 

to the apoE genotype on neurodegeneration. Wild and transgenic 

mice expressing human apoE3 or apoE4 are exposed to different 

diet conditions such as a normal diet, high saturated fats and 

cholesterol intake. After 6 months of diet mice will be tested to 

evaluate cognition and memory impairments. In the same subjects, 

gene expression (cDNA microarrays) will be studies in discrete brain 

areas. RT-PCR assay will be used to confirm the results. Moreover, 

selected genes will be further analyzed with classical biochemical 

procedures. In addition, other AD proteins such as Aβ, APP and Tau, 

and several markers for apoptosis, necrosis and neurodegeneration, 

will be investigated with biochemical methods in order to determine 

their quantity, their activity and their distribution in the brain. The 

combination of high throughput methods with the classical 

biochemical assays will allow a deep understanding of the molecular 

mechanisms behind the studied risk factors involvement in 

neuropatology in relation with apoE genotype. The obtained results 

will provide important information to define life guidelines for 

people carrying the apoE4 allele, as prevention for AD and 

neurodegenerative disorders. This could be of relevance in public 

health strategies. Furthermore, results could help to identify new 

therapeutic targets and early diagnostic markers. Moreover, this 

new experimental approach will develop a new animal model for AD 

that has a more natural onset of the pathology.

Nome 

Rosa Maria Corbo 

Contatti Rosamaria.corbo@uniroma1.it 

Istituto/Dipartimento Dip. Genetica e Biologia Molecolare, Università La Sapienza, Roma 


Sporadic Alzheimer’s disease (AD) is a complex neurodegenerative disease caused by 

the interaction of genetic and nongenetic factors. The e * 4 allele of the apolipoprotein E gene 

(APOE) is the only well-established genetic susceptibility factor for sporadic AD, though 

numerous risk alleles in other genes are also thought to be involved in AD development. 

Gender is a relevant AD risk determinant and there is evidence for a higher prevalence of AD 

in women. However, whether this is due to the longer life expectancy of women or to 

biological gender- specific risk factors for the disease is unclear. It has been hypothesized that 

a declining estrogen level following menopause and subsequent reduction in its 

neuroprotective action may contribute to the development of the disease in women. In line 

with this hypothesis, the possible role of estrogen nuclear receptor genes (ESR1 and ESR2) as 

AD risk factors has been largely investigated. In addition there is evidence that past fertility is 

associated with a higher AD risk and a reduced AAO of the disease, probably because of the 

lifelong decrease in estrogen levels that parity seems to produce in women. 

The aim of the present investigation is to extend previous investigations (1-3) and in 

particular 1) to collect data on the reproductive life of AD women to identify those 

parameters which could influence the risk of AD and/or AD onset age; 2) To study the genetic 

variability of genes involved in estrogen metabolism or women fertility. 

- Corbo RM, Gambina G, Ruggeri M, Scacchi R. Association of estrogen receptor α (ESR1) 

PvuII and XbaI polymorphisms with sporadic Alzheimer’s disease and their effect on 

apolipoprotein E concentrations. Dement Geriatr Cogn Disord, 22:67-72, 2006- 

- Combined effect of Apolipoprotein E (APOE) genotype and past fertility on age at onset of 

Alzheimer’s disease in women. Dement Geriatr Cogn Disord, 24: 82-85, 2007 

- Genetic variation of CYP19 (aromatase) gene influences age at onset of Alzheimer’s 

disease in women. Dement Geriatr Cogn Disord. 2009, 27, 513-518. 




Genetic susceptibility to Alzheimer’s disease 

Alzheimer’s disease in women: AD as a gender disease 

A. Basic research 

Investigation on common and rare variation of genes involved in sporadic 

Alzheimer’s Disease susceptibility 

Ente finanziatore Università di Roma La Sapienza 



At present the genetic component of the sporadic 

late-onset form of Alzheimer disease (LOAD) has been only 

partially elucidated, since only the apolipoprotein E (APOE) 

gene, specifically the e*4 allele, has been reportedly 

associated with the disease. Since the APOE polymorphism 

alone does not entirely explain the genetic predisposition to

sporadic AD, investigation into whether any variation in 

further candidate genes could account for the remaining part 

may offer important clues. However, investigations on AD 

candidate genes have usually examined only common 

polymorphisms in accordance with the hypothesis “common 

disease /common variants” . The additional hypothesis 

suggesting the presence of a relevant genetic heterogeneity 

due to rare alleles (“common disease/rare variants” ) has 

been scarcely investigated . The aim of the present study 

was 1) to examine the common variation of numerous 

candidate genes (ACT, PSEN1, LDL-R, APOC1, alfa2M, PON1, 

LPL, AChE, BChE, Chat, CYP19); 2) to carry out a mutation 

screening in the exons of APOE, PSEN1, APP genes by means 

of DHPLC (denaturing high performance liquid 

chromatography). (product: 12 publication, see pubmed)

Nome Prof. LUCIA MIGLIORE 

Contatti 

Istituto/Dipartimento University of Pisa 

Department of Human and Environmental Sciences 

Via S. Giuseppe, 22 

56026 PISA Italy 

E-mail: l.migliore@geog.unipi 

Fax: +39 050 2211034 

Tel: +39 050 2211029-28 






- Genetic susceptibility to Alzheimer’s disease 

(Epigenetic marks of susceptibility to Alzheimer disease) 

Abstract 

AD is characterized by high Hcy and low folate blood levels. Many of the 

genes participating in folate metabolism are polymorphic, meaning that 

this pathway can be altered by low dietary folate intake, by the 

polymorphic variants and/or by combinations of both. In vitro studies 

showed that in conditions of altered folate metabolism the status of 

methylation of the promoter of the PSEN1 gene underwent a variation, 

causing a deregulation of presenilin 1, BACE and APP proteins. 

The study is designed to evaluate the contribution of folate metabolism 

(including polymorphisms of metabolic genes and plasma levels of folate, 

homocysteine (Hcy) and VitB12) in both early (MCI) and late stages of 

dementia, and its possible contribution to the methylation status of the 

promoter of genes that participate in Aβ processing. The study will be 

performed in blood samples with the goal of identifying easily detectable 

epigenetic biomarkers of disease susceptibility. 

A PILOT STUDY ON THE EFFECTS OF FPP ON THE COGNITIVE FUNCTION 

AND ON OXIDATIVE STRESS PERIPHERAL BIOMARKERS IN 

MILDCOGNITIVE IMPAIRMENT PATIENTS (MCI) 

OSATO Research Institute, Gifu, Japan 

Durata progetto Project ongoing 


30+30 amnesic MCI patients divided into two main groups. One treated 

with fermented papaya preparation (FPP), the other without treatment

(placebo: PLC). 

The study will be performed within 12 months for a total treatment time 

of six months. After six and twelve months a report will analyze the 

results. At the beginning of the pilot study all the subjects recruited will 

be screened for the Total Antioxidant Status (TAS) that will include the 

determination of the following values in blood: selenium, folic acid, 

vitamin B12, C, E together with the evaluation of the GSH/GSSG ratio. The 

groups will be analyzed for assessing primary and oxidative DNA damage 

levels (Comet assay) and chromosome damage also due to oxidative 

damage (Cyt-B micronucleus assay) in peripheral blood leukocytes. 

Moreover 8OH-dG levels in urines samples will be evaluated. 

We plan also to evaluate the following biochemical parameters of 

oxidative stress: advanced oxidation protein products (AOPP); 

glutathione; hydroxynonenal; lipoperoxide (basal and under stress 

conditions); lactate (basal and under stress conditions); mitochondrial 

metabolite (by Mass Spectroscopy). 

The decrease (if any) of oxidative stress biomarkers due to the FPP 

subministration will be evaluated by comparing after12 months the two 

MCI groups (FPP treated and placebo controls) data by means of 

statistical analyses. At the same time the cognitive impairment evolution 

will be monitored by means of the neuropsychological evaluations also 

within the treatment period (0, 6 and 12 months). Moreover for all the 

subjects involved, DNA extraction will be performed and the samples will 

be stored for further genotype analysis (e.g.: ApoE, ..).

Nome LAURA COLOMBAIONI 

Contatti laura.colombaioni@in.cnr.it> 

Istituto/Dipartimento Istituto di Neuroscienze CNR, Pisa 



identificata 


Role of altered lipid and cholesterol metabolism in neurodegenerations 

Titolo progetto Cellular mechanisms of neuronal dysfunction due to altered lipid metabolism. 




Associazione Italiana Niemann-Pick 

3 years 

Deregulated lipid metabolism is of particular importance in pathogenesis of CNS disorders 

and injuries. For this reason the role of cholesterol and lipid metabolism as susceptibility 

factors in neurodegenerations needs to be further investigated. Cholesterol is an 

important regulator of lipid organization in neuronal membranes and is the precursor for 

neurosteroid biosynthesis. Low levels of neurosteroids are related to poor outcome in 

many brain pathologies. The deregulation of the principal cholesterol carrier protein in 

the brain, the Apolipoprotein E, is a significant risk factor for Alzheimer's disease. 

Similarly, Parkinson's disease is, in some degree, caused by an enhancement of lipid 

peroxidation. Also Niemann-Pick A-B diseases are caused by sphingomyelin accumulation, 

while Niemann-Pick disease C is due to mutations in either the NPC1 or NPC2 genes, 

resulting in defective cholesterol and sphingolipid transport. Sphingolipid metabolites are 

emerging as members of a novel class of lipid mediators, acting both as intracellular 

second messengers and as ligands of cell surface receptors, capable of controlling 

neuronal proliferation, differentiation and apoptosis. 

In my laboratory, the effects, on neuronal cells, of an altered lipid 

metabolism are examined. The goal is to understand how the altered lipid 

metabolism produces neuronal injury, what subcellular compartments are 

involved and how this occurs. An interdisciplinary approach is used employing 

cell biology, molecular biology and live cell imaging methods. By using confocal 

microscopy, some key events associated to altered cholesterol trafficking or to 

sphingolipid unbalance are characterized, such as mitochondrial deregulation, 

release of pro-apoptotic effectors and alterations of calcium homeostasis in 

specific subcellular compartments. 

In particular, the effects on neuronal calcium homeostasis of some sphingolipid 

metabolites such as Glucosyl-ceramide Sphingosine and Sphingosine 1-phosphate 

are analyzed. Our results indicate that the calcium balance in mitochondria and 

in Endoplasmic Reticulum is regulated by cholesterol and sphingolipid 

metabolites and that this is critically involved in control of neuronal physiology. 

Understanding the impact of lipid metabolism changes in neuronal cell functions will 

promote novel concepts and will reveal new therapeutic targets for a variety of nervous 

system disorders.

Nome Roberto Cosimo Melcangi 

Contatti 

Tel. 

E mail 

02-50318238 

roberto.melcangi@unimi.it 

Istituto/Dipartimento DEFIB-Università degli Studi di Milano 


Area di interesse identificata F. Endocrine and metabolic factors 



Sex-specific therapeutic strategies based on neuroactive steroids for 

Alzheimer's disease. 

Ente finanziatore partial support by Bayer-Pharma 


Abstract del progetto The importance of sex in the incidence of Alzheimer's disease (AD) has 

been studied. Indeed, epidemiological studies support a higher 

prevalence and incidence of AD in women. Moreover, as observed in 

several experimental models and in AD patients, parameters such as 

oxidative damage, plaque load, amyloid precursor protein, β-amyloid 

production, seizure activity, locomotion, hippocampal 

neurodegeneration, astrocyte proliferation, level of brain-derived 

neurotrophic factor, cognition are differently affected in a sex-dimorphic 

way. Although sex differences in the brain may be in part the 

consequence of the genetic complement of the cell, differentiation of the 

neural tissue is also influenced by the developmental actions of sex 

hormones. Both genetic sex and developmental actions of sex hormones 

contribute to the generation of sexually dimorphic neuronal networks in 

the brain and the spinal cord, and both factors may influence the 

outcome of pathological insults to the nervous system. Thus, some sex 

differences in brain pathology may be the consequence of the functional 

and morphological differences in neural structure between the sexes. 

However, it is important to remember that CNS is not only a target for 

steroid hormones coming from peripheral glands but also of those 

directly synthesize in loco (i.e., neurosteroids). Moreover, steroid 

hormones are also actively CNS converted into metabolites that are more 

active and in some cases utilize a different mechanism of action. Both 

steroid hormones and neurosteroids may be included in the neuroactive 

steroid family. 

Therefore, the assessment of the levels of neuroactive steroids directly in 

male and female brain areas of experimental models of AD might be 

extremely important. Using our previous experience in other 

experimental models of neurodegeneration, we intend to evaluate the 

levels of neuroactive steroids by liquid chromatography tandem mass 

spectrometry and to confirm possible changes analyzing by real time PCR 

the gene expression of molecules and enzymes involved in their 

synthesis. These observations might be very useful to identify possible 

pharmacological target useful to design sex-specific therapeutic 

approaches based on neuroactive steroids.

Nome Roberto Maggi 

Contatti 

Tel. 

E mail 

02 50318233 – 3293718540 

roberto.maggi@unimi.it 

Dipartimento Endocrinology, Physiopatology, Applied Biology 


Area di interesse identificata Selective Alzheimer Diserase Indicator-1 (Seladin-1): a linker between 

cholesterol, oxidative stress and Alzheimer’s disease. 



Ente finanziatore PRIN – MIUR 2006069900_002 



Alzheimer's disease: correlation with the Gonadotropin Hormone 

Releasing Hormone (GnRH). 

Amyloid beta (Abeta) plaques represent one of the primary pathological 

features that appear in the brains of Alzheimer's patients. Recent findings 

suggest that the complete proteolytic machinery required for Abeta 

generation is located within lipid rafts. Membrane cholesterol content is 

crucial for lipid raft organization and it seems to regulate the membrane 

localization and the activity of secretases involved in the metabolism of 

APP. Epidemiological and biochemical studies indicate that all changes in 

hypothalamic-pituitary gonadal (HPG) hormones (sex steroids, LH, 

Gonadotropin releasing Hormone or GnRH) levels following 

menopause/andropause are involved in the cognitive and 

neuropathological changes observed in AD. Sex steroids have been shown 

to be potent neuroprotective agents and a decreased incidence and delay 

in the onset of AD has been described among women on hormone 

replacement therapy (HRT). 

Direct roles of GnRH in AD have received scant attention, even if phase III 

clinical trials are currently underway to establish the role of a GnRH 

agonist in lowering the incidence of dementia and in improving cognitive 

functions of AD patients. 

Recently, it has been discovered that estrogens are able to up-regulate 

the new gene seladin-1 (Selective Alzheimer disease indicator 1). Seladin- 

1 is a gene down-regulated in the brain areas involved in AD and its overexpression 

confer protection against Abeta mediated toxicity. Seladin-1 

was found to be identical to DHCR24, the enzyme that catalyzes the last 

step of cholesterol biosynthesis. Therefore seladin-1 could represent a 

key regulator of membrane cholesterol and it could be the molecular 

mediator of the neuroprotective effect of estrogens. This project is 

devoted to study the role of two hormones of the HPG axis, estrogen and 

GnRH, in AD pathogenesis. Moreover, we will investigate the molecular 

mechanisms that can link these hormonal actions with the cholesterol 

content of plasma membrane and with the cholesterol-rich microdomains 

organization and functionality, and finally the possible role for seladin-1 

as a molecular mediator of this biological mechanisms. For this research 

proposal we will utilize different appropriate humane (FNC-B4 and SH- 

SY5Y) and murine (GT1-7, GN11, and primary hippocampal neurons) 

cellular models.

The experimental plan aims to clarify three points: 

1.The involvement of membrane cholesterol and lipid membrane 

organization in the processing of the APP and its modulation by 

estrogens. We will understand if estrogen stimulation is able to alter the 

organization of membrane lipids and proteins and the proteolytic 

processing of the APP, and if these modifications correlate to changes of 

the cholesterol content and of seladin-1 expression. 

2.The role of GnRH in the modulation of the cytotoxic action of Abeta and 

the APP processing and in neuronal excitability. We will clarify how GnRH 

affect APP processing, Abeta deposition and Abeta toxicity and how GnRH 

modulate excitability in hippocampal neurons. 

3.The hormonal regulation of seladin-1, its ability to modulate the HPG 

hormone action and its involvement in Abeta induced cytotoxicity. We 

will evaluate the role of seladin-1, as a regulator of membrane 

cholesterol, in the amyloidogenic pathway, in the amyloid-induced 

neurotoxicity and in lipid rafts organization and functionality as "signaling 

platforms". 




PRIN – MIUR 2006069900_002 

PUR – MIUR 2008 


Abstract del progetto Alzheimer's disease: correlation with the Gonadotropin Hormone 

Releasing Hormone (GnRH). 

Amyloid beta (Abeta) plaques represent one of the primary pathological 

features that appear in the brains of Alzheimer's patients. Recent findings 

suggest that the complete proteolytic machinery required for Abeta 

generation is located within lipid rafts. Membrane cholesterol content is 

crucial for lipid raft organization and it seems to regulate the membrane 

localization and the activity of secretases involved in the metabolism of 

APP. Epidemiological and biochemical studies indicate that all changes in 

hypothalamic-pituitary gonadal (HPG) hormones (sex steroids, LH, 

Gonadotropin releasing Hormone or GnRH) levels following 

menopause/andropause are involved in the cognitive and 

neuropathological changes observed in AD. Sex steroids have been shown 

to be potent neuroprotective agents and a decreased incidence and delay 

in the onset of AD has been described among women on hormone 

replacement therapy (HRT). 

Direct roles of GnRH in AD have received scant attention, even if phase III 

clinical trials are currently underway to establish the role of a GnRH 

agonist in lowering the incidence of dementia and in improving cognitive 

functions of AD patients. 

Recently, it has been discovered that estrogens are able to up-regulate 

the new gene seladin-1 (Selective Alzheimer disease indicator 1). Seladin- 

1 is a gene down-regulated in the brain areas involved in AD and its overexpression 

confer protection against Abeta mediated toxicity. Seladin-1 

was found to be identical to DHCR24, the enzyme that catalyzes the last 

step of cholesterol biosynthesis. Therefore seladin-1 could represent a 

key regulator of membrane cholesterol and it could be the molecular 

mediator of the neuroprotective effect of estrogens. This project is

devoted to study the role of two hormones of the HPG axis, estrogen and 

GnRH, in AD pathogenesis. Moreover, we will investigate the molecular 

mechanisms that can link these hormonal actions with the cholesterol 

content of plasma membrane and with the cholesterol-rich microdomains 

organization and functionality, and finally the possible role for seladin-1 

as a molecular mediator of this biological mechanisms. For this research 

proposal we will utilize different appropriate humane (FNC-B4 and SH- 

SY5Y) and murine (GT1-7, GN11, and primary hippocampal neurons) 

cellular models. 

The experimental plan aims to clarify three points: 

1.The involvement of membrane cholesterol and lipid membrane 

organization in the processing of the APP and its modulation by 

estrogens. We will understand if estrogen stimulation is able to alter the 

organization of membrane lipids and proteins and the proteolytic 

processing of the APP, and if these modifications correlate to changes of 

the cholesterol content and of seladin-1 expression. 

2.The role of GnRH in the modulation of the cytotoxic action of Abeta and 

the APP processing and in neuronal excitability. We will clarify how GnRH 

affect APP processing, Abeta deposition and Abeta toxicity and how GnRH 

modulate excitability in hippocampal neurons. 

3.The hormonal regulation of seladin-1, its ability to modulate the HPG 

hormone action and its involvement in Abeta induced cytotoxicity. We 

will evaluate the role of seladin-1, as a regulator of membrane 

cholesterol, in the amyloidogenic pathway, in the amyloid-induced 

neurotoxicity and in lipid rafts organization and functionality as "signaling 

platforms".

Nome Laboratory of Biochemistry & Molecular Biology of Lipids - Mass 

Spectrometry "Giovanni Galli" 

Contatti 

Tel. 

E mail 

Donatella Caruso, Maurizio Crestani, Emma De Fabiani, Nico Mitro 

0250318344 – 0250318393 

donatella.caruso@unimi.it, maurizio.crestani@unimi.it, 

emma.defabiani@unimi.it, nico.mitro@unimi.it. 

Istituto/Dipartimento Department of Pharmacological Sciences 


Area di interesse identificata Nuclear receptors, coregulators, neuroactive steroids and epigenetics in 

neurodegenerative diseases associated to metabolic disorders. 







G. Cell interactions

Nome Prof. Ubaldo ARMATO, MD 

Contatti ubaldo.armato@univr.it 

Istituto/Dipartimento Histology & Embryology Unit, Dept. of Life and Reproduction Sciences, 

University of Verona Medical School, Verona, Venetia, I-37134, Italy 


The Astrocyte, not just a Bystander in Alzheimer’s Disease 

(in collaboration with Balu Chakravarthy e James F. Whitfield, Institute 

for Biological Sciences, National Research Council of Canada, Ottawa, 

Ont., K1A 0N6, Canada) 

Area di interesse identificata - Multidisciplinary projects 


- Basic research 

Titolo progetto --- 

Ente finanziatore --- 

Durata progetto ---- 

Abstract del progetto The Astrocyte, not just a Bystander in Alzheimer’s Disease 

There are two kinds of Alzheimer’s disease (AD). There is the rare early 

onset AD (EOAD) which includes less than 5% of the cases and is due to 

mutation-induced hyperproduction of the Aβ42 (amyloid β-(1-42) 

fragment of the membrane-associated amyloid precursor protein (APP). 

Then there is the slowly developing, late-onset AD (LOADAD that includes 

95-99% of the cases. The mutant driver genes of EOAD have been 

relatively easy to identify. But the key LOAD drivers have been far harder 

to find in the host of changes associated with the progressive breakdown 

of neuronal-glial circuits starting in the entorhinal cortex and spreading 

through the hippocampus and into the cerebral hot-spots of the cerebral 

DMN (default mode neuronal network ) ending in catastrophic cognitive 

failure with the loss of memory formation by the hippocampus being an 

outstanding feature. So far the attention of workers in the AD field 

focused on neurons. 

But things are changing. Now we know that certain (radial) astrocytes are 

the stem cells for adult neurogenesis in the hippocampal dentate gyrus 

and therefore are key components of the memory-forming machinery. 

We are learning that massive astrocyte syncytial networks connect to, 

and collaborate with, neurons in sustaining and driving brain functions. 

And they are co-conspirators with neurons in the cognitive catastrophe of 

AD. Therefore we have developed a method of culturing normally 

functioning (minimally altered) astrocytes directly following their removal 

from adult human cerebral cortices. We are using these cells, which we 

call NAHAs (normal human astrocytes), to explore the possible roles of 

astrocytes in AD. There is an ever-escalating list of things that might cause 

of help cause LOAD. At first it was thought that the striking fibrillar 

amyloid plaques in AD brains were the neuronal circuit breaking killers. 

But plaque-rich brains of cognitively normal or even above-normal 90-

years old nuns and the failure of plaque formation in transgenic mice to 

correlate with neuronal and cognitive loss argue that this is probably not 

true. It is now suspected that it is a build-up of invisible pre-plaque Aβ42 

oligomers that start destroying synapses and disconnecting cognitive 

circuits. We have been and will keep working on three things that seem 

to be part of the LOAD process: the induction of VEGF production by 

astrocytes; the expression and function of p75 NTR , an Aβ42 and 

neurotropin receptor. 

VEGF Production We have so far found that both Aβ42 and a 

combination of three glial-induced cytokines (IL-1β, IFN-γ and TNF-α) that 

appear in the inflammatory environment of an AD brain can induce 

NAHAs to produce and secrete mainlythe VEGF-A165 isoform. Interestingly 

none of the three cytokines can significantly stimulate VEGF production, 

but together they are more potent stimulators than Aβ35-42 (an active 

fragment of Aβ42). We must now find out how Aβ42 (i.e., Aβ35-42) by 

itself and how the three cytokines together produce the HIF-1α•HIF-1β 

heterodimer that stimulates VEGF production in the NAHAs. Is VEGF just a 

minor player in AD? What might VEGF do in an AD brain? The release of 

angiogenic VEGF from the Aβ42/cytokine ensemble-stimulated astrocytes of 

the neurobarrier-coupling units with their end-feet attached to blood vessels 

damaged by deposited Aβ42 would be expected to reverse, or at least 

minimize, the Aβ42-induced vascular damage widely believed to be 

responsible for hypoxia, glucose shortages, breached blood-brain barrier and 

failing Aβ42 drainage. But then it may not. Hypoxic endothelial cells in the 

hypoperfused regions of an AD brain downregulate their MEOX-2 homeobox 

gene and its GAX protein product. This causes the endothelial cells to 

upregulate the AFX1 forkhead transcription factor that downregulates their 

anti-apoptogenesis Bcl-XL gene by stimulating the BCL-6 transcriptional 

repressor. The downregulation of MEOX-2 and its GAX protein product also 

lowers the level of the Aβ42-clearing LRP1. Therefore while the increased 

VEGF production in an AD brain would indeed stimulate angiogenesis, the new 

blood vessels would be dysfunctional with endothelial cells unable to clear 

Aβ1-42 and hyper-prone to apoptogenesis. But VEGF may have another 

impact on memory formation. It is a major factor in the maintenance of the 

vascular-based the radial glial (astrocyte-like) neuronal stem cell niche in the 

dentate gyral subgranular zone, a center of adult neurogenesis. VEGF 

stimulates the production of the niche’s endothelial cells that produce the 

neurostimulatory BDNF factor. Thus, angiogenesis is linked to neurogenesis 

in the niche to continuously supply the new granular cells needed to start 

the recording the memory of novel polymodal inputs from the entorhinal 

cortex. Therefore the increased expression of VEGF in a normal brain or 

injection of VEGF into such a brain might promote the survival and 

proliferation of neural progenitor cells as indicated by the reported 

response of hippocampal neuronal progenitor cells to infusion of the 

protein into the lateral ventricles of adult rats. But this may be unlikely in an

AD brain even with its elevated VEGF expression, because it has been 

reported that the core AD pathology prevents any VEGF-stimulated 

immature granule cells from differentiating into mature granule cells. 

p75 NTR , the villainous Darth Vader of the AD brain? p75 NTR combines with 

the TrkA neurotrophin receptor to drive brain development when activated 

by BDNF and NGF. But it fades away after development only to re-emerge 

in the aging, Aβ42 –accumulating AD brain along with the downregulation 

of TrkA. Now without restraint by TrkA it becomes dangerous because it 

has a cytoplasmic apoptogenic DD (‘death domain’) which is turned on 

when the receptor is activated by Aβ42 binding to its neurotrophin binding 

site. When its DD is not opposed by associated by TrkA and it meets Aβ42 

on a neuronal surface it kills the cell via its DD. We have found that 

accumulating Aβ42 actually stimulates the expression of p75 NTR by 

neurons and NAHAs. Obviously this could accelerate AD development. 

Aβ42 and neurotropin receptor. But then there are also indications of a 

Aβ42/ p75 NTR vicious cycle. Activated p75 NTR receptors have been shown 

to stimulate caveolar sphingomyelinase to release ceramide from a 

pool of sphingomyelin in the cell membrane. The released ceramide 

post-transcript-ionally stabilizes BACE1 which, by β-cleaving membrane- 

associated AβPP, accelerates the drive to full-blown AD by increasing Aβ42 

production. This Aβ42would then switch back and bind to p75 NTR receptors 

and increase the calpain-driven cleavage of the receptor’s C-terminal 

membrane-associated domains which ultimately induce the receptors’ 

cytoplasmic death domains and trigger a cytocidal apoptogenic caspase 

8/caspase 3 cascade. We are now trying to confirm the contrapuntal rise 

and fall of p75 NTR and TrkA in a key region of the human AD brain such as 

the hippocampus. And along with this we are trying to find out with 

whether NAHAs can start an Aβ42/ p75 NTR vicious cycle. If they can, it will be 

a strong indication that astrocytes have a major role in the cognitive 

collapse of the AD brain.

Nome Manuela MARCOLI 

Contatti 

Dipartimento di Medicina Sperimentale (DIMES), Sezione di Farmacologia 

e Tossicologia – Università degli Studi di Genova 

Viale Cembrano, 4 - I-16148 Genova, Italy 

Phone: +39 010 3532656; Fax: +39 010 3993360 

E-mail: marcoli@pharmatox.unige.it 

Istituto/Dipartimento Dipartimento di Medicina Sperimentale (DIMES), Sezione di Farmacologia 

e Tossicologia – Università degli Studi di Genova 

The research strategy we propose addresses the need for better knowledge of early molecular events 

leading to neuronal dysfunction in Alzheimer’s disease (AD). We propose to focus on the effects of soluble 

amyloid-beta peptides (Aβ) on mechanisms sustaining neuron-astrocyte cross-talk at glutamatergic 

synapses and on network synaptic activity in Multi Electrode Array (MEA)-coupled neuron/astrocyte 

cultures, in both wild type and triple transgenic AD mouse (3xTg-AD mouse). Combined use of isolated 

models (nerve terminals and astrocyte processes) and an integrated model (MEA-coupled 

neuron/astrocyte cultures) will provide complementary data to understand Aβ-related dysruption of 

glutamatergic transmission in AD. Assessment of changes of synaptic transmission at the level of astrocyte 

processes and nerve terminals (exposed to Aβ) together with analysis of the effects of (endogenously 

produced and exogenously added) Aβ on neuron/astrocyte network activity may shed new light on the 

molecular and cellular mechanisms underlying early dysfunction of glutamatergic transmission in AD, 

helping to discover new therapeutic targets to improve nerve cell signal transduction and ultimately 

preventing neuronal dysfunction. 

Background and rationale: Astrocytes regulate neuron function through nonoverlapping domain 

organization: one astrocyte processes contact thousands of synapses, promoting neuronal synchrony and 

regulating synaptic transmission; the neuro/gliotransmitter glutamate mediate bidirectional neuronastrocyte 

communication. Although altered glutamate transmission and astrocyte global perturbation are 

reported in AD, scarce attention is focused on the potential role of astrocyte processes in synaptic 

dysfunction in AD. Soluble Aβ appears involved in synaptic dysfunction; focus on the impact of Aβ on 

mechanisms sustaining neuron-astrocyte cross-talk at glutamatergic synapses might unravel key steps in 

synaptic impairment and in the cascade of events causing specific sets of neurons becoming dysfunctional. 

Experimental models and Methods: Aβ effects on [Ca 2+] i and transmitter release will be directly 

investigated on astrocyte processes (gliosomes) and nerve terminals (synaptosomes) from adult wild type 

and 3xTg-AD mouse brains. Gliosomes and synaptosomes stem from astrocytes and neurons obtained 

from adult mouse brain; in situ maturation of neurons and astrocytes would assure that the impact of 

intracellularly accumulating Aβ would also be accounted for when studying glutamate release and Ca 2+ 

signaling in nerve terminals and astrocyte processes prepared from 3xTg-AD mouse brain. Integrated 

effects of Aβ on network activity and synaptic connection maturation will be analyzed by non-conventional 

electrophysiology on cultured neurons/astrocytes from wild type and 3xTg-AD mouse brain on MEAs, 

allowing long-term, non invasive, multielectrode detection of neuronal network activity and 

synchronization; the bioelectronic system has proven very sensitive in detecting early neuron damage and 

loss of synaptic transmission. 

Human resources: key persons, experience and know-how that can be accessed without additional funds: 

Manuela Marcoli, Guido Maura (University of Genova, Department of Experimental Medicine DIMES; 

Center of Excellence for Biomedical Research CEBR; National Institute of Neuroscience INN): transmitter 

release from CNS preparations (nerve terminals, astrocyte processes) and cell cultures 

Sergio Martinoia (University of Genoa, Department of Biophysical and Electronic Engineering DIBE; 

Italian Institute of Technology IIT): Micro electrode arrays technology; MEA-based systems and in-vitro 

neural interface and neuro-robotics 

Mario Nobile (National Research Council, Institute of Biophysics, Genova): [Ca 2+ ]I imaging on single nerve 

terminals and astrocyte processes 

Area di interesse identificata First challenge: scientific; understanding the mechanisms of the disease

Priorites: - Basic research 

- Multidisciplinary projects 

- New treatment strategies 

Finanziamenti ricevuti - ongoing research support 

Titolo progetto Neuronal differentiation: functional evaluation of neurotransmission 


MIUR – PRIN mod B Anno 2008 - prot. 20088JEHW3_005 

(to Guido Maura) 


Abstract del progetto To understand the molecular mechanisms associated to the 

differentiation of neuron precursor cells, we plan to use Neuroblastoma 

N18TG2 clones, transfected with Choline Acetyl Transferase (ChAT) and 

overexpressing early growth response factor-1 (EGR-1), that mimic 

neuron precursor cells. The research program plans to investigate on the 

possibility to induce a fully developed neuron phenotype in N18TG2 

clones transfected with ChAT and/or overexpressing EGR-1, by inhibiting 

the expression of the Repressor element-1 Silencing Transcription (REST), 

coding for a transcriptional repressor of neurogenesis. Our analyses will 

focus on evaluation of the cell ability to communicate through 

neurotransmitters and regulated exocytosis upon REST knock down; if 

this occurs the gene expression profile will be determined. The project 

will reveal gene networks relevant for complete maturation of the 

neuroblastoma cell line, and hopefully will be of help in understanding 

the neuron maturation process. 

Finanziamenti ricevuti - ongoing research support 


Mechanisms of glial and neuronal glutamate release as specific 

therapeutic targets in brain ischemia 


Fondazione Cassa di Risparmio di Genova, 2008 

(to Mario Nobile, role of MM: participant) 


Abstract del progetto The aim of the project is to get new light on molecular and cellular 

mechanisms activated during/after ischemia to find out pharmacological 

targets for neuroprotection. Brain ischemia results from multiple related 

mechanisms involving both neurons and glial cells. The neuron-glia crosstalk, 

subserved by mechanisms including glutamate is of fundamental 

importance to the synapse physiology and to the synaptic transmission 

dysregulation in neurodegenerative diseases; increased extracellular 

glutamate and intracellular calcium overload lead to cell death. Multiple 

mechanisms are responsible for glutamate release: vesicular release, 

glutamate transporter reversal, and possibly activation of ATP 

P2X7receptors, of volume-regulated anion channels or of 

connexin/pannexin. hemichannels. These ways for glutamate release can 

function both on neurons and astrocytes, although the role for glial cells 

in ischemic neuron damage is still debated. As the pharmacological 

intervention based on glutamate ionotropic receptor antagonism was 

unsatisfactory, knowledge of the glutamate modes of exit during ischemia 

might help to find new specific therapeutic targets. The research strategy 

will be based on the investigation at synapse level, at the level of single 

cell and of the cell network (slices), on the participation of P2X7 

receptors, VRAC channels and pannexin in ischemic damage in vitro. As 

we previously found that adenosine controls astrocyte P2X7 receptors, 

we will investigate on the relationship between adenosine and P2X7 

receptors during ischemic insult.

Finanziamenti ricevuti - submitted proposal 


Synapse and Amyloid-beta: effects on astrocyte processes and nerve 

terminal 


Alzheimer’s Association 2010 Investigator-Initiated Research Grant 

Proposal Identifiers LOIID: 173512: approved 

Full proposal: under evaluation 

(role of MM: principal investigator) 



Understanding of neuron-astrocyte interaction at synapses is crucial to 

successful therapeutic approach to neurodegenerative diseases. 

Astrocytes regulate neuronal function through nonoverlapping domain 

organization: the processes of one astrocyte contact thousands of 

synapses, promoting neuronal synchrony and regulating neuron 

excitability and synaptic transmission. The neuro/glio transmitters 

glutamate and ATP mediate bidirectional neuron-astrocyte 

communication at synapses. Although altered glutamate or ATP 

transmission and astrocyte global perturbation are reported in AD 

models, scarce attention has been focused on the potential role of 

astrocyte processes in synaptic dysfunction in AD. Soluble amyloid-beta 

peptides are involved in synaptic dysfunction, an early event in the 

pathogenesis of AD before synapse loss. Our proposal focus on the effects 

of soluble amyloid-beta on the mechanisms subserving bidirectional 

communication between astrocyte processes and nerve terminals. Focus 

on the impact of amyloid-beta peptides on astrocyte process might 

unravel key steps in synaptic impairment and in the cascade of events 

causing specific sets of neurons becoming dysfunctional. How? Astrocyte 

processes (gliosomes) and nerve terminals (synaptosomes) from wild type 

and transgenic AD mouse brains will be used, allowing direct investigation 

on the amyloid-beta effects on intracellular calcium and transmitter 

release. It is worth noting that gliosomes and synaptosomes stem from 

astrocytes and neurons obtained from adult mouse brain; in situ 

maturation of gliosome-producing astrocytes and of synaptosomeproducing 

neurons provides experimental models suitable to investigate 

on amyloid-beta effects in adult life. Integrated effects of amyloid-beta on 

network activity and synaptic connection maturation will be analyzed on 

co-cultured neurons/astrocytes from wild type and transgenic AD mouse 

brain on multielectrode arrays, by non-conventional electrophysiology 

(allowing long-term, non invasive, multielectrode detection of 

spontaneous and evoked firing activity and neuronal network 

synchronization). This bioelectronic system has proven very sensitive in 

detecting early neuronal damage and loss of synaptic transmission. Why? 

Assessment of changes of synaptic transmission at the level of astrocyte 

processes and nerve terminals (exposed to amyloid-beta) together with 

analysis of the amyloid-beta effects on neuron/astrocyte network activity 

may shed new light on the molecular and cellular mechanisms underlying 

early neurodegenerative processes and cognitive dysfunction in AD, 

helping to discover new therapeutic targets to improve nerve cell signal 

transduction and ultimately preventing neuronal dysfunction and synapse 

loss.

Finanziamenti ricevuti - completed research support 


Feasibility of a prototypical system development for in vitro testing for 

biomedical, toxicological and ambiental purpose 


Scientific and Technological Park of Liguria Program Objective 2 2000- 

2006 (to Sergio Martinoia, role of MM: responsible of Unit) 


Abstract del progetto The goal of this project was to set up neuron cultures on multielectrode 

arrays allowing neuropharmacological/neurotoxicological analysis of the 

integrated effects of neuroactive substances on network activity 

Combining the study of the network firing activity with the study of the 

release of the excitatory transmitter glutamate and of the activation of 

the intracellular pathways following ionotropic receptor (NMDA, 

AMPA/kainate) activation and involved in the excitotoxic neuron cell 

damage or death would allow an integrated mechanistic-based approach 

to neurotoxicity evaluatio in neuron network. Non conventional 

electrophysiological and neurochemical approach allow 

neuropharmacological /neurotoxicological analysis of the integrated 

effects of neuroactive substances on the network activity and dissection 

of the mechanisms underlying complex network responses to the 

substances. 



Pharmacological control of extracellular excitatory amino acid 

accumulation and cellular calcium overload in neonatal 


ischemia/reperfusion 

Mariani Foundation (Milan) R-07-66 

(to Mario Nobile, role of MM: participant) 


Abstract del progetto The study evaluates acute and long-term (60 days) effects of perinatal 

hypoxia, in order to find new pharmacological targets to be used in 

intervention devoted to prevention/protection against neonatal ischemic 

neuron damage. Perinatal hypoxia-ischemia is a major cause of neonatal 

morbidity and mortality. A number of evidences indicate that ischemic 

neuron damage can depend on increase of extracellular K + accumulation 

of excitatory aminoacids and of other transmitter (such as 

adenosine/ATP), and intracellular Ca 2+ overload, triggering excitotoxicity 

and apoptotic/necrotic nerodegeneration. Such events might play 

different roles depending on the developmental stage of central nervous 

system. Scarce reports are available on specific models, allowing 

dissection of diverse mechanisms possibly involved and temporal 

sequences of neonatal ischemia. Relative contributions of neurons and 

glia and of their bi-directional communication in the control of glutamate 

synaptic level and in excitotoxicity triggering and execution, are not 

clearly defined. Knowledge of the modes for efflux of excitatory 

aminoacid and their regulation, in early and late phases of 

ischemia/reperfusion, could open the possibility to reduce the 

extracellular excitatory aminoacid levels in different neonatal ischemia 

phases. As intracellular Ca 2+ accumulation seems prerequisite for neuron 

damage cascade, dampening of Ca 2+ influx through ionotropic glutamate

or purinergic receptors (e.g. P2X7) could significantly reduce neuron 

damage. We therefore propose investigation on glutamate efflux and 

cellular Ca 2+ levels in physiological conditions and during ischemia in vitro 

models of neonatal brain (preparations from neonatal and 60 days-old 

rats, models for premature infant and adulthood brain, respectively). 

Parallel studies will be conduced with the aim to unravel whether 

glutamate release from astroglial cells can be modulated by endogenous 

signaling molecules through the regulation of swelling-activated Cl - 

channels. New pharmacological targets to be used in intervention 

devoted to prevention/protection against neonatal ischemic neuron 

damage, will be searched for through the following intermediate steps: - 

better knowledge of the functional role played by ionotropic glutamate 

and purine receptors on neuron and glial cells; - better knowledge of the 

modes responsible for extracellular glutamate accumulation and of their 

relative importance in the temporal sequence of events during 

ischemia/reperfusion; -better knowledge of the mechanisms regulating 

activity of swelling-activated Cl - channels involved in the pathologic 

release of excitatory amino acids; - identification of receptors whose 

activation (or blockade) might affect vesicular excitatory aminoacid efflux 

or glutamatergic pathways or neuron-glia activation in different phases of 

ischemia/reperfusion. 



Neuron networks coupled in vitro to microtransducers and artificial 

body: a new potent device to study neuronal interfaces and cerebral 

plasticity in vitro 


Italian Ministry of University and Scientific Research (FIRB 2002) 

RBAU012KF8_005 

(to Guido Maura, role of MM: participant) 


Abstract del progetto The goal of the project was to investigate on spontaneous and 

electrically-evoked activity of neuronal networks on microtransducer 

arrays for a better knowledge of the cell mechanisms underlying burst 

generation and propagation. To this end, non conventional 

electrophysiology on MEA will be applied to detect the effects of 

chemical interference on neuron-specific features linked to neuron 

functioning (electrophysiological events at level of single cells and at 

the network level) and will be combined with neurochemical analysis 

for neuron-specific features linked to neuron functioning 

(neurotransmitter processing and release; presence and functioning of 

neurotransmitter receptors and linkage to transduction mechanisms) 

combined with the electrophysiological analysis.

Nome Domenici Luciano 

Contatti domenici@in.cnr.it; luciano.domenici@univaq.it 

Istituto/Dipartimento Neuroscience Institute, CNR, Via G. Moruzzi 1, 56100 Pisa; Dep. STB, 

University of L’Aquila, L’Aquila. 


Area di interesse identificata Alzheimer’s disease, Ageing 



Active projects: 

1) RAGE, MAP kinases and Abeta induced synaptic dysfunction. 

2) From brain ischemia to beta amyloid toxicity in Alzheimer’s 

disease; neurobiological mechanisms, new biomarkers and 

therapeutical approach. 

Submitted projects: 

- Vascular triggering of beta amyloid toxicity in AD. 

- BDNF and beta amyloid toxicity, building a bridge from 

neurobiological mechanisms to treatment of brain disorders 




1) American Health Assistance Foundation (AHAF). 

2) Regione Toscana, Regional Health Research Program 2009 (first 

step approval passed, final approval and funding pending). 

1) Two years. 

2) Two years. 

1. Abstract project AHAF. 

Beta amyloid protein (Aβ) is the principal component of senile plaques, a 

neurophatological prominent feature of Alzheimer's disease (AD). 

Increasing evidence suggests that loss of neuronal function and cognitive 

impairment precede the accumulation of Aβ plaques in AD. We 

hypothesize that oligomeric Aβ is involved in neuronal dysfunction during 

an early phase of AD. We will investigate the mechanisms underlying 

oligomeric Aβ induced synaptic dysfunction in entorhinal cortex (EC) 

whose impairment contributes to cognitive decline in AD. We will address 

three specific aims: 

1. To test the hypothesis that Aβ affects cortical synaptic plasticity. The 

effects of different concentrations of oligomeric Aβ on synaptic 

transmission and efficacy will be investigated in EC slices using 

extracellular and single cell recordings. 

2. To verify the identity of receptors and intracellular pathways 

transducing Aβ signal. Receptor for Advanced Glycation Endproducts 

(RAGE) is a cell surface binding site for Aβ. We will investigate the effects 

of oligomeric Aβ on synaptic transmission and efficacy in EC slices: i) in 

the presence of anti-RAGE antibodies, ii) in the transgenic (Tg) mice with 

targeted neuronal expression of the wild type form of RAGE and in RAGEdeficient 

mice; iii) in the Tg mice with a dominant negative form of RAGE 

targeted to neurons (DN-RAGE). Finally, we will investigate the 

involvement of different kinases associated with RAGE activation, such as 

p38 MAPK, p42-p44 MAPK and JNK. 

3. To test the role of neuronal RAGE in vivo using animal models of Aβmediated 

synaptic dysfunction. We will employ Tg mice overexpressing 

mutant human amyloid precursor protein (mAPP); Aβ-rich environment in 

Tg mAPP mice induces synaptic dysfunction. We expect that synaptic 

dysfunction observed in Tg mAPP mice does not occur in Tg mAPP mice 

crossed with Tg DN-RAGE mice. Finally, we will determine whether

increased expression of neuronal RAGE in Tg mAPP mice exaggerates 

synaptic dysfunction. 

2. Abstract Project Regione Toscana. 

Alzheimer’s disease (AD) is a progressive neurodegenerative disorder of 

the elderly characterized by senile plaques composed of beta amyloid 

(Aβ), neurofibrillary tangles, synaptic loss and impairment of cognitive 

functions. AD is the most common cause of dementia followed by 

vascular dementia (VaD). The incidence of neurodegenerative disorders in 

AD is increased by vascular pathologies such as stroke, clogging arteries 

and hypertension inducing brain hypoperfusion and hypoxia. However, a 

causal relationship between AD and brain ischemia at the molecular level 

has not been established. The present project will contribute to elucidate 

whether Aβ/RAGE signalling may be viewed as part of a neural system 

that controls synaptic responses in vascular pathology and AD. We are 

interested in investigating this issue as many patients with AD also have 

hypertension and vascular infarctions. Designing new biomarkers of 

neurodegeneration and interfering with Aβ/RAGE signalling is expected to 

have high impact to ameliorate cellular dysfunction in AD and vascular 

pathology. Patentability of a new diagnostic method of AD based on 

biomarkers associated with RAGE activation and deposition of Aβ 

represents the goal of our plan.

Nome Evelina Chieregatti 

Contatti evelina.chieregatti@iit.it 

Istituto/Dipartimento Dept. of Neuroscience and Brain Technologies, IIT, Genoa, Italy 


Synaptic deficits in neurodegenerative diseases 

The main lines of our research concern the investigation of the early mechanisms underlying synaptic 

dysfunctions in Parkinson’s and Alzheimer diseases. These neurodegenerative diseases are characterized 

by the deposition of proteinacious aggregates in the neurons and in the brain tissue. 

However it has been shown that most neurons that undergo cell death do not contain aggregates and 

that, whether they contain aggregates or not, are similarly affected by morphological dendritic 

abnormalities or biochemical changes. It may thus be conceivable that, rather than being caused by the 

accumulation of aggregates with neuronal cell bodies, which could be an epiphenomenon, 

neurodegenerative diseases may be characterized by dying back mechanisms that begin at the synapse 

and lead to axonal degeneration. Both β amyloid (Aβ) and α-synuclein (Syn) are pathogenetic proteins that 

upon mutations or increase in their levels may induce alterations in the synaptic function and neuronal cell 

death. A key element in the regulation of neurotransmitter release and in the fine tuning of synaptic 

efficacy is the neuronal cytoskeleton. Both proteins are implicated in actin dynamics as well as microtubule 

stability through their interactions with a microtubule-binding protein, tau. We recently demonstrated a 

disrupting effect of the mutated form of Syn on actin dynamics in neurons. We are studying their effects 

on the morphology and dynamics of actin microfilaments and intermediate filaments by the use of purified 

proteins and neurons derived from knock-out and transgenic mice. The long term goal of this project is to 

investigate the effect of the two proteins on cytoskeletal-regulated processes, such as polarization, 

regeneration after axotomy and neuronal plasticity. Another project concerns the characterization of the 

molecular interactors of Syn that may have a role in neurodegeneration associated with αsynucleopathies. 

The intracellular localization of the cleavage of APP by BACE1 has long been a topic of debate. Early 

endosome is the first site of APP endocytic sorting, where APP has been shown to co-localize with BACE1. 

Altered endosomal structures have been found in AD brains. Our aim is to identify proteins of interest such 

as enzymes or molecular motors that may be found in the APP-containing endocytic vesicles. 

It is expected that the results will shed light on the role of Syn and Aβ in synaptic function, and on the 

molecular interactors mediating their effect, providing cues useful to reach appropriate therapeutic 

endpoints based on the modulation of the signalling pathways involved in the pathogenic effect of Syn and 

Aβ. 



Macromolecular interactions and Neurodegeneration 


The role of intraneuronal amyloid β(Aβ) peptide on the pathogenesis of 

the Alzheimer disease: functional and proteomic analysis. 

Ente finanziatore Fondazione CARIPLO – role: Partner 

Durata progetto 2 years (end 2009) 

Abstract del progetto Alzheimer disease (AD), one of the neurodegenerative diseases of the 

nervous system characterized by a chronic process of cell death, is the 

most frequent form of dementia, but there is still no therapeutic 

intervention capable of preventing, delaying or treating it. Intraneuronal 

Aβ accumulation precedes the deposition of amyloid plaques and the


appearance of neurofibrillary tangles, and correlate with the first 

appearance of cognitive deficits. The role of intraneuronal Aβ in the AD 

pathogenesis is still unclear, but recent data suggest a possible 

involvement in cytoskeletal functions and neurotransmitter release. The 

goal of this project is to understand how Aβ may interfere with synaptic 

function and, by the use of advanced proteomic techniques, to identify 

the proteins that interact with Aβ. The results we expect to obtain will 

shed light on the pathogenetic mechanisms in AD and will allow the 

development of advanced diagnostic and therapeutic protocols. 

Pathogenetic mechanisms of familial Parkinson’s disease: wt and A30P 

α-synucleins affect the structure of microfilaments and intermediate 

filaments. Pathways and effects on cytoskeletal dynamics. 

Ente finanziatore Telethon-Italy – role: PI 

Durata progetto 3 years (end 2013) 

Abstract del progetto Overall objectives. To understand a physiological role of alpha-synuclein 

(Syn) and its role in the pathology of Parkinson's disease (PD) by 

comparative studies of wt and A30P Syn mutant interaction with 

microfilaments and intermediate filaments. To understand Syn role in 

cellular processes that require active remodeling of these two 

cytoskeletal elements. 

Specific aims. Analysis of the regulation of actin-Syn interaction in living 

cells by fluorescence energy transfer (FRET). Analysis of the composition 

of actin/A30P Syn aggregates formed during cytoskeleton remodeling. 

Analysis of Syn effects on intermediate filaments turnover and on 

neuronal regeneration after axotomy. Investigation of the pathway(s) 

that mediates the effect of extracellular Syn on actin cytoskeleton 

morphology. 

Background. Mutations in the Syn gene are responsible for familial PD. 

Syn knock-out mice suggest a role of Syn in the regulation of the presynaptic 

vesicular pools in neurons. Syn has been involved in neuronal 

regeneration occurring during differentiation and repair. Interestingly, 

actin microfilaments and the intermediate filaments are actively involved 

in every one of these processes. 

Description of the project. Based on our data demonstrating interaction 

between Syn and actin we propose to study the regulation and the 

subcellular localization of their interaction by FRET. In neurons in culture 

from knock-out and from conditional transgenic mice we plan to 

investigate the role of Syn in regeneration after axonal injury. We 

propose to investigate the transduction pathway that is activated by 

extracellular Syn. 

Anticipated output. We expect to identify the specific step at which Syn 

function at the pre-synapse and the possible loss of function of the A30P 

Syn mutant. These results will be of great relevance in the understanding 

of PD pathogenesis.


H- Intracellular Traffiking

Nome Cecilia Bucci 

Contatti 

Tel. +39-0832-298900; Fax +39-0832-298626 

e-mail: cecilia.bucci@unisalento.it 

Istituto/Dipartimento Department of Biological and Envirnmental Sciences and Technologies 

(DiSTeBA), University of Salento 


Area di interesse identificata Membrane traffic and neurodegenerative diseases: role of Rab proteins 

and their effectors 

The laboratory of Applied Biology directed by Cecilia Bucci is focused on 

the role of endocytic Rab proteins. Rab proteins are small GTPases that 

control membrane traffic in the endocytic and exocytic pathways. Indeed, 

they are responsible, through the recruitment of several specific 

effectors, for several aspects of membrane transport as vesicle formation, 

vesicle movement onto cytoskeletal tracks, vesicle docking and tethering 

to the target compartment, and, finally, vesicle fusion with the target 

compartment. There are about 70 rab genes in the human genome. 

Alterations of membrane traffic, and, in particolar, of Rab proteins or of 

their effector proteins, are responsible for many human genetic and 

acquired diseases including cancer. In particular, Rab dysfunction has 

been linked to several neurodegenerative diseases. For instance 

mutations in Rab7 protein cause the Charcot-Marie-Tooth type 2B 

disease, a inherited neuropathy of the peripheral nervous system. Late 

endocytic dysfunction has been demonstrated in Alzheimer’s disease and 

presenilin (a protein mutated in Alzheimer) interacts with the endocytic 

Rab11 and with the Rab regulatory protein GDI (GDP-dissociation 

inhibitor). In addition, presenilin influences localization and function of at 

least other two Rab proteins, Rab6 and Rab8. Rab6 expression is 

increased in Alzheimer’s disease brain. Mutations in alpha-synuclein are a 

cause of familial Parkinson disease; mutated alphasynuclein interacts 

abnormally with Rab3a, Rab5 and Rab8. 

These are only some examples that demonstrate the close connection 

between membrane traffic (and in particular Rab proteins function) and 


We propose therefore to study different aspects of membrane traffic at 

the molecular level, with a particular focus on Rab proteins, in order to 

understand the molecular mechanism underlying degenerative diseases 

as Alzheimer, Parkinson, Charcot-Marie-Tooth, etc. In particular, we 

could investigate the role of the proteins found mutated in these diseases 

(i.e. presenilin, alpha-synuclein, Rab7, etc.) in membrane traffic, and the 

role of their interaction with regulators of membrane traffic. 


Titolo progetto Molecular basis of Charcot-Marie-Tooth type 2B disease 

Ente finanziatore Telethon Italy 

Durata progetto 3 years (2009-2012) 

Abstract del progetto The inherited neuropathies of the peripheral nervous system show 

clinical and genetic heterogeneity. CharcotMarieTooth type 2B (CMT2B) is 

a ulceromutilating neuropathy clinically characterized by prominent 

sensory loss, marked distal muscle weakness and wasting, high frequency 

of foot ulcers, infections, and amputations of the toes because of 

recurrent infections. Four missense mutations in the small GTPase late 

endosomal protein Rab7 have been identified in autosomal dominant

ulceromutilating neuropathy families. These missense mutations cause 

the CMT2B phenotype and target highly conserved amino acid residues. It 

is not known how these mutations affect specifically peripheral neurons 

leading to an axonal pathology in CMT2B. This issue is particularly 

challenging considering that Rab7 is a ubiquitous protein. This proposal 

plans to continue investigating the 

molecular mechanism responsible for the development of the disease. 

We have previously biochemically characterized the Rab7 mutated 

proteins causative of CMT2B in HeLa cells, determining that they are 

mechanistically similar. Indeed, we have demonstrated that activated 

forms of Rab7 are responsible for the disease.Within this project we plan 

to: 

i) do functional studies using the mutated proteins on PC12, SH−SY5Y, 

Neuro2a cells and neurons ii) screen DRG and motor neuron cDNA 

libraries with different methods to search for new Rab7 interacting 

proteins, looking in particular at effectors expressed specifically in the 

peripheral nervous system. iii) do functional assays to establish the role of 

these newly identified Rab7 interacting proteins and to establish if and 

how Rab7 mutations affect their function. Understanding how 

dysfunction of Rab7 causes CMT2B will be instrumental to gain insights in 

the pathogenesis of this group of disorders, and will open the way to 

future identification of therapeutic approaches.

Nome Prof. Paolo Macchi, PhD 

Contatti Phone: +39 0461-883819/883095 

Fax: +39 0461-883937 

Email: macchi@science.unitn.it 

Istituto/Dipartimento Molecular and Cellular Neurobiology Lab 

CIBIO – Centre for Integrative Biology, University of Trento. 

Via delle Regole 101 

3060 Mattarello, Trento – Italy 

WEB: http://www.unitn.it/cibio 

Research proposal 

“Comparative analysis of the transport and translational RNPs complexes in neurons: an integrate 

transcript- and proteomic approach.” 

RNA localization is emerging as an important process to restrict certain proteins to specific subcellular 

domains and thus spatially control the expression of genes within cells. One key feature of messenger RNA 

(mRNA) localization is that it precedes translation. As a consequence, mRNA has to be kept translationally 

silent during its transport towards the proper target compartment. Localized mRNAs are packaged into 

ribonucleoprotein particles (RNPs), composed of RNAs and proteins. The localized RNAs contain specific 

cis-acting sequence elements, which are recognized by proteins, called trans-acting factors. The 

importance of RNA subcellular localization in the formation and function of the nervous system is also 

gaining increasing acceptance. In highly polarized cells, such as in neurons, a functional 

compartmentalization is crucial. RNA localization is not only involved in establishing the structural polarity 

of neurons, but is also assumed to play an important role in synaptic plasticity 1 . It has been estimated that 

approximately 200 different mRNAs encoding for proteins important for synaptic plasticity as well as 

various housekeeping genes, are found in dendrites of hippocampal neurons 2 . Interestingly, the loss-offunction 

of one component of the RNA transport machinery and/or the misexpression of specific mRNAs 

causes abnormalities during neuronal development. For example, defects in axonal and dendritic 

outgrowth and/or in dendritic spine development have been described. These phenotypes have recently 

been correlated with different neurological disorders, such as FXS (fragile X mental retardation syndrome), 

SCA (spinocerebellar ataxia) and SMA (spinal muscular atrophy) 3 . However, the composition of the 

transport as well as the translation control machinery is still poorly characterized. Our research focuses on 

dissecting the molecular composition of the neuronal transport RNPs under physiological as well as 

pathological conditions (e.g. neurodegeneration, brain tumours and epilepsy). 

In detail, we are trying to answer the following biological questions: 

1 Is the local protein synthesis and/or mRNA trafficking altered in neurons undergoing degeneration? 

2 What is the molecular composition of the neuronal RNPs (RNA and protein interactome) in neurons 

affected by degeneration compared to normal cells? 

The goal of purifying transport RNPs to identify novel components has been previously attempted, but has 

not yielded the precise mechanistic details of the process that are sought. Our laboratory has successfully 

demonstrated the feasibility of maintaining endogenous transport complexes containing Staufen, Barentsz 

and Pumilio proteins, RNA (RIP-chip analysis), and motor proteins in cell-free extracts of brains and 

cultured neurons 4,5 . Using our collection of affinity-purified antibodies to immunoprecipitate localization 

RNPs from brain extract fractions, the composition of RNPs isolated from wild type mice will be assessed 

and compared with RNPs isolated from animal model for neurological diseases. We will then test the 

individual functions of both known (or suspected) and novel identified components of neuronal RNPs in a

variety of in vivo and in vitro assays to characterize their roles in localization and associated mechanisms. 

Finally, since the activation of translation of dendritically localized mRNAs upon specific inputs, such as 

synaptic activity, allows for the modulation of existing synapses, notably the morphological and functional 

changing of dendritic spines, 

3 Are there possible drugs able to affect mRNA targeting (and/or turnover) and local protein synthesis 

that can be used to block neurodegeneration? 

We are currently setting up a high throughput drug screening assay on neuronal cells at CIBIO (University 

of Trento). RIP-Chip experiments will be functional for the development of pharmacological assays where 

the level of expression of mRNAs identified will be evaluated through the use of a reporter protein 

(luciferase). Through a bioluminescence assay we will be able to quantify the effect of each chemical's 

inhibitory activity or promoter of the complex proteins/ mRNA-luciferase. 

We propose that a multidisciplinary study to underline the mechanisms that lead to correct regulation of 

both RNA and RNA-binding proteins will be crucial for a better understanding of the pathogenesis of 

neuronal diseases. Interdisciplinary internal collaborations, with other institutions in Italy and abroad have 

been already established to take full advantage of different experimental systems and areas of expertise. 

1. Kuhl D. & Skehel P. (1998) Curr. Opin. Neurobiol. 8, 600-606. 

2. Job C. & Eberwine J. (2001) Nat. Rev. Neuroscie. 2, 889-898. 

3. Dahm R. & Macchi P. (2007). Biol. Cell 99, 649-61. 

4. Mallardo M. et al., (2003). PNAS 100, 2100-2105 

5. Vessey J. et al., (2010). PNAS 107, 3222-3227 

Area di interesse identificata Basic research. Neurodegenerative diseases. 


Titolo progetto Electrophysiological characterization of alpha-synuclein forming channels, 

a protein involved in the neurodegeneration of Parkinson's disease 

Ente finanziatore CARITRO Foundation 


Abstract del progetto The aggregation of aberrant proteins is involved in the pathogenesis of 

neurodegenerative diseases associated with aging but the mechanisms 

leading to neuronal death are unknown. The α-synuclein (αS) is the main 

component of Lewy bodies, namely fibrillar intraneuronal inclusions 

present in all patients with Parkinson's disease (PD), and is strongly 

indicated as the cause of the disease. The αS is not structured in a 

"native" form while in vitro form amyloid fibrils that are rich in beta 

sheet. Nell'αS mutations found in patients with PD promote the 

formation of annular and tubular structures of protofibrille. The 

protofibrille of αS, but not the monomer or mature fibrils, bind to 

structures such as lipid vesicles, planar and cellular membranes. Here, 

they assume a new structural conformation similar to the pores formed 

by the bacterial toxins that cross the membrane. These "amyloid pores" 

may be the cause of neuronal death, through an uncontrolled and 

nonspecific permeabilization of the cell membrane. We intend to 

investigate in detail the electrophysiological characteristics of the pores 

of αS formed in model systems of increasing complexity: in lipid planar 

membranes, synaptic vesicles from liposomes and/or mitochondria and 

eventually in cultured neurons. In particular an innovative analysis 

technique suitable for noise characterization of pores will be developed. 

We intend to investigate, with techniques of analysis of noise, EEG from

healthy patients with PD and to highlight similarities with anomalies 

recorded in single cells. We believe that this multidisciplinary approach 

will allow 'early diagnosis and more precise onset and evolution of PD as 

well as of other neurodegenerative diseases.


I- Animal models

Nome Adalberto MERIGHI 

Contatti 

Phone:++390116709118 

Fax: ++390112369118 

e-mail: adalberto.merighi@unito.it 

Istituto/Dipartimento Istituto Nazionale di Neuroscienze e Dipartimento di Morfofisiologia 

Veterinaria, università degli Studi di Torino 

Via Leonardo da Vinci 44 

I-10095 GRUGLIASCO (TO) Italy 


Effects of over-expression of the anti-apoptotic proteins BCL-2 and survivin on the endoplasmic 

reticulum stress induced by β amyloid: study of neuroprotection on animal models 


Developing competitive animal models to study Alzheimer’s disease 

The etiology and mechanisms responsible for neurodegeneration in Alzheimer’s disease (AD) are 

multifaceted, but recent studies indicate that activation of an intrinsic gene-regulated program of cell 

death (PCD = programmed cell death) is a common end-point of the various intracellular pathways leading 

to neuronal loss. Multiple intracellular pathways activated during PCD, but, among them, apoptosis plays a 

major role. The post-natal cerebellar cortex is an ideal model for the study of PCD, because of the massive 

apoptosis of the granule cells, a population of cerebellar excitatory interneurons formed by billions of cells 

(see for example Lossi L. and Merighi A. Prog Neurobiol. 2003. 69:287-312). β-amyloid (Aβ) is directly 

involved in neurodegeneration observed in AD, where neurodegenerative changes are due, at least in part, 

to activation of an apoptotic programme (see for example Wisniewski T. Neurobiol Dis. 1997. 4:313-28). In 

primary cortical neuron cultures, the toxic effects of synthetic peptides corresponding to specific 

sequences of Aβ have been shown to be consequent to release of calcium across ryanodine receptor (RyR) 

channels and/or inositol-3-phosphate channels (IP3) on the membrane of the endoplasmic reticulum 

(Ferreiro E. Neurobiol Dis. 2006. 23:669-78). We have recently demonstrated that the block of RyR 

channels in organotypic cultures obtained from mouse cerebellum interferes with the increase in density 

of cerebellar granule cells expressing the BCL-2 protein (one of the most widely characterized antiapoptotic 

factors in Eukaryotic cells) fused with a fluorescent reporter marker (EYFP = enhanced yellow 

fluorescent protein) after KCl depolarization (Lossi et al. Dev Neurobiol. 2009. 69:855-870). These 

observations suggest that BCL-2 could modulate the apoptotic program triggered by Aβ as a consequence 

of endoplasmic reticulum stress. 

Survivin is one member of a family of proteins commonly referred to as apoptosis inhibiting proteins. It is 

expressed during mitosis, and in tumors, but also during normal development of neurons in vivo, as 

demonstrated in conditional transgenic mice using the Cre-loxP technology (Jiang Y. et al. J Neurosci. 2005. 

25:6962-70). In these animals the conditional deletion of the survivin gene at P10.5 leads to massive 

apoptosis in several areas of the central nervous system. As a consequence, newborn mutants display a 

marked reduction of brain size, with severe multifocal apoptosis affecting the cerebral and cerebellar 

cortices, the brainstem , the spinal cord and the retina. Caspase 3 and 9 (the main effector molecules of 

apoptosis)activities are increased, whereas expression of the anti-apoptotic protein BAX is unaltered. 

This project consists of two phases carried out in sequence. The first phase will consist of: 

- Pharmacological manipulation of organotypic cultures by addition to culture medium of Aβ peptide 

fragments; 

- Qualitative and quantitative evaluation of the effects the fragments after labeling of live/dead cells by

fluorescent probes (Syto10/Dead Red, Invitrogen, USA or iodidium propide); 

- Immunocytochemical analysis of apoptosis with special reference to activation of caspases/PARP1 and 

cell cycle pathways (retinoblastoma protein, chk1, cdc2) 

- Analysis of relationship between proliferation and apoptosis following visualization of proliferating cells 

with immunofluorescence (H3 histone, BrdU) and apoptotic cells with the TUNEL technique. In parallel 

cleavage of apoptosis related proteins will be analyzed by Western blotting. 

The second phase of the project will consist of: 

- Evaluation of the effects of Aβ after biolistic transfection of cultures with the following vectors: ApoAlert 

Caspase 3 sensor (Clontech, USA) to monitor the activation of caspase 3 in alive cells; pEYFP-N1 (Clontech, 

USA) with insertion of the cDNA for human BCL-2; pHcRed-C1 (Clontech, USA) with insertion of the cDNa 

for survivin (Obtained from Dr. Rachel Altura, Ohio, USA); 

- Analysis of apoptosis and cell cycle kinetics in transfected cells transfection (as for control cultures 

above); 

- Analysis of cell calcium dynamics by real time calcium imaging. 

Specific experimental procedures 

Gene gun technology: We will employ a biolistic transfection procedure (gene gun) to artificially introduce 

one or more genes of interest in tissue slices. The procedure involves shooting of slices with gold 

microcarriers coated with cDNAs of different constructs encoding for molecule(s) under study and for a 

reporter protein (GFP/EYFP/DsRed). Gold microcarriers will be accelerated into tissue by a helium blast 

(180 psi) so that they will penetrate cell membranes and reach the cytoplasm of target cells. Transfection 

will be performed after 2-5 days in vitro, by a commercial apparatus (Helios Gene Gun System, BioRad, 

USA). 

Confocal microscopy: Control and transfected cultures will be incubated inside a custom made microscope 

stage incubator (Tokai Hit, Japan) and monitored at variable intervals of time under a Leica SP5 laser 

confocal microscope. We have already verified that by this set up it will be possible to monitor the cultures 

for 4 days or longer under the microscope, and it will be thus possible to study short to medium term 

effects on apoptosis. In certain experiments a cell permeant calcium indicator will be used (Oregon Green 

Bapta 1AM, Invitrogen, USA)to monitor the intracellular calcium concentration in individual neurons. 

Visualization of proliferating cells: a technique based upon incorporation of bromodeoxyuridine (BrdU) 

during S phase will be used by adding BrdU directly to culture medium. M and G2 phase cells will be 

labeled by immunocytochemistry using a primary antibody directed against phosphorylated H3 histone. 



Effects of over-expression of the anti-apoptotic proteins BCL-2 and 

survivin on the endoplasmic reticulum stress induced by prion protein 

and βamyloid: study of neuroprotection on animal models 

Ente finanziatore MIUR PRIN 2007 (to Laura Lossi) 

Durata progetto 24 mesi

Abstract del progetto Several neurodegenerative diseases are characterized by an initial 

endoplasmic reticulum stress followed by the activation of an apoptotic 

program leading to cell death and neuronal loss. 

The two main goals of this project are: 

- analysis of apoptosis induced by prion protein (PrP) e da β-amyloid (Aβ) 

in an ex vivo model consisting of murine organotypic cerebellar cultures 

obtained from P7-P20 mice. This developmental stage has been proved to 

be particularly suitable to our purposes since it is characterized by a 

massive apoptotic neuronal death. 

- analysis of potentially neuroprotective effects of two anti-apoptotic 

proteins (BCL-2 and survivin) onto neuronal death induced by PrP and Aβ. 

The project has duration of 24 months, and is organized into two phases, 

each corresponding to one of the two main objectives outlined above. 

We will employ a multidisciplinary approach consisting of in vitro 

culturing techniques, biolistic transfection (gene-gun), confocal 

microscopy (medium to long term analysis of transfected cell, reat time 

imaging of calcium influx), in situ immunocytochemistry and molecular 

biology (immunofluorescence, labeling of proliferating/apoptotic cells), 

biochemistry (Western blotting). 

All these procedure will eventually aim to obtain a direct correlation 

between morphological and functional data. 

We expect to obtain a better characterization of the basic biology of PrP 

and Aβ in relation to modulation of programmed cell death. 


Development of an in vitro system to study the neuroprotective effects 

of ghrelin and its derivatives on central neurons 

Ente finanziatore MIUR PRIN 2008 

Durata progetto 24 mesi 

Abstract del progetto Background and rationale: Ghrelin is a peptide hormone produced by the 

endocrine cells of the gastric mucosa. Its most relevant biological action is 

the regulation of feeding behavior in the presence of a negative energetic 

balance. After discovery at periphery, the hypothalamus has been 

identified as the main source of ghrelin in the central nervous system 

(CNS). Recent studies show that the hormone has indeed a wide range of 

biological effects in central neurons (see Ferrini et al. 2009, Current 

Neuropharmacology. 7:37). Among these, we have recently characterized 

the role of ghrelin in spinal cord pain neurotransmission (Vergnano et al. 

2008. Endocrinology. 149:2306). However, data have been obtained in 

support to the notion that ghrelin also plays a role in neuronal 

proliferation ,differentiation, protection against apoptosis and 

maintenance of a differentiated status (see Chung et al. 2007. 

Endocrinology. 148:148). To understand whether or not the effects of 

ghrelin in cultured cells are indeed of relevance in vivo it is necessary to 

respond to the following question: is the hormone produced in the 

stomach capable to cross the blood brain barrier? The importance of this 

question appears immediately obvious if one considers that the 

physiological levels of ghrelin expression in CNS are relatively low. 

General layout and timetable of project: This is a two year project. 

During the first year we will first test the antiapoptotic effects of ghrelin 

and its derivatives on organotypic cerebellar slices. Then we will develop 

an in vitro model based on our previous experience (postnatal cerebellar 

slices cultured in vitro under static conditions - see Lossi et al. 2009. Prog 

Neurobiol. 88:221) where the integrity of the blood brain barriers is

maintained. This model will be derived according to a recently developed 

protocol (Bendfeldt et al. 2007. J Neurosci. 27:3260) where it was 

demonstrated that the cultivation of cortical slices in the presence of 

FGF2 (basic fibroblast growth factor 2) is ideal to the preservation of the 

blood brain barrier. During the second year of the project we will assess 

the effects of ghrelin on cell death in relation to integrity of the blood 

brain barrier. Aims and methods: The project focuses on the study of the 

antiapoptotic effects of ghrelin in a widely established model of 

developmental cell death, the post-natal cerebellar cortex. The effects of 

the hormone will be compared in the presence/absence of the blood 

brain barrier to understand whether or not peripheral ghrelin is capable 

to influence the survival of central neurons. To do so we will use a 

multidisciplinary approach consisting of in vitro cultures, cell transfection 

(gene-gun), confocal microscopy (mid-to-long term analysis of cell 

calcium fluxes), immunocytochemistry, molecular biology in situ 

(immunofluorescence, labeling of proliferating/dead cells), and 

biochemistry (Western blotting). The concurrent employment of all these 

procedures will allow obtaining a direct correlation between 

morphological and functional data. Project phases: 1. Development of 

FGF2 stimulated cerebellar organotypic cultures and assessment of 

integrity of the blood brain barrier. 2. Pharmacological manipulation of 

cultures with ghrelin, agonists and antagonists. 3. Analysis of cell death by 

means of fluorescent probes (Syto 10/Dead Red, Invitrogen, USA). 4. 

Immunocytochemical analysis of the molecules involved in the apoptotic 

cascade (retinoblastoma protein, cell cycle-related kinases chk1, cdc2), 5. 

Analysis of the correlation between apoptosis and cell proliferation (after 

labeling with fluorescent probes and/or specific markers). 6. Western 

blotting of relevant proteins to study their cleavage in the course of cell 

death.

Nome Monica DiLuca 

Contatti 

Universita’ degli Studi di Milano 

Via Balzaretti, 9 

02 50318374 

monica.diluca@unimi.it 

Istituto/Dipartimento CEND and Dept of Pharmacological Sciences 





Synaptic dysfunction in neurodegeneration: characterization of gene 

products in functional synaptic networks under physiologicaò and 

pathological conditions. This includes characterization of mechanisms 

involved in subsynaptic targeting of proteins and protein complexes. 

These studies will allow for identificationa nd validation of targets as 

molecules responsible for synaptopathies using pharmacological 

approaches, siRNA, mimetic peptides and small synthetic molecules. In 

addition, these approaches will lead to model stages of 

neurodegenaration in vivo by obtaining innovative animal models for 

Alzheimer diseases and other neurodegenerative disorders. 

Role of calcium dependent gene expression and protein trafficking in 

shaping post synaptic activity: implication for neurodegenerative 

diseases. 

Ente finanziatore Cariplo foundation 

Durata progetto April 2009 – March 2011 

Abstract del progetto the main goal of the project is to investigate the link between 

dyshomeostasis of calcium signalling and early pathogenic events of AD – 

both at cellular and molecular level - working on the hypothesis that this 

neurodegenerative disorder is a synaptopathy. As an addendum to this, 

we do not wish to focus on the role of calcium in neuronal loss but on the 

role of initial small changes of calcium homeostasis in synaptic deficits, as 

in vivo this correlates better with early neurological disturbances and 

cognitive impairment. 

Titolo progetto Molecular mechanism and behavioural aspects in experimental models of 

Parkinson’s disease. 

Ente finanziatore Italian Minister of Health 

Durata progetto November 2008 – November 2010 

Abstract del progetto Despite abundant evidences support the role of NMDA receptor complex 

in Parkinson’s Disease, only scattered if any information are available at 

single subunit level, although this is a much-needed information in order 

to identify possible new molecular pathways that can represent a 

pharmacological target for the disease as well as strategies to tackle 

dyskinesia. Thus, specific aim of the project will be to characterize in 

striatal neurons the molecular events responsible for the correct synaptic 

localization of NR2A and NR2B subunits of NMDA receptor and to study 

the functional consequences of such an event in experimental 

parkinsonism and in L-DOPA induced dyskinesia. 

Titolo progetto “REPLACES” - Restorative Plasticity At Corticostriatal Excitatory Synapses 

Ente finanziatore EU VII Framework Programme 

Durata progetto November 2008- October 2012 

Abstract del progetto The main ideas underlying this proposal is that the long-term efficacy of

new treatments for Parkinson’s disease (PD) will be conditioned by their 

ability to restore, both structurally and functionally, the “synaptic wiring” 

of striatal neurons and physiological synaptic plasticity. Accordingly, the 

proposal will address the potential restorative effects of either novel 

pharmacological treatments or neuronal transplants on the corticostriatal 

microcircuitry. Since chronic treatment with the DA precursor L-DOPA 

induces in the majority of PD patients a maladapTative plasticity causing 

involuntary movements (dyskinesia), innovative strategies should prevent 

the development of this disabling condition. The project will characterize 

corticostriatal synaptic plasticity from molecular aspects to clinical 

neurophysiology involving also behavioural as well as morphological 

analysis of the basal ganglia system. 


“cPADs” - Cell permeable peptides as drug delivery system: a way 

towards innovative therapeutic strategies for neurodegeneration. 

Ente finanziatore EU VII Framework Programme 

Durata progetto April 2008- March 2012 

Abstract del progetto The project aims to foster a dynamic pathway between academic 

research organizations and private commercial enterprises, based on a 

long-term co-operation programme in the field of innovative therapeutic 

strategies for neurodegenerative disorders. Through inter-sectoral 

industry-academia collaboration, the proposal aims to define appropriate 

technologies to target key protein-protein interactions and key 

intracellular pathways mediating neurodegeneration and cell death, 

addressing both the creation of innovative models and the design of 

innovative drugs by means of exploiting the use of Cell-Permeable 

Peptides.

Nome Prof. Renata Bartesaghi 

Contatti 

Tel +39 051 2091727, +39 051 2091747 

Fax +39 051 2091737 

e-mail: renata.bartesaghi@unibo.it 

Istituto/Dipartimento Dipartimento di Fisiologia Umana e Generale, Università di Bologna 


Study of early onset Alzheimer disease in a model for Down syndrome, the Ts65Dn mouse 

Area di interesse identificata: 

DEVELOPING COMPETITIVE 

ANIMAL MODELS TO STUDY 

ALZHEIMER’S DISEASE 


BASIC RESEARCH 

Individuals with Down syndrome (DS) develop neuropathological 

features similar to Alzheimer disease (AD) early in life, including 

dementia, accumulation of beta-amyloid, and irregular 

phosphorylation of tau proteins. Ts65Dn mice have an extra copy of 

the distal aspect of mouse chromosome 16, a segment which is 

homologous to human chromosome 21 and which contains much of 

the genetic material responsible for the DS phenotype. Ts65Dn mice 

show developmental delay during the postnatal period as well as 

abnormal behaviors in both young and adult animals that may be 

analogous to mental retardation. Though the Ts65Dn brain is 

normal on gross examination, there is age-related degeneration of 

septohippocampal cholinergic neurons and astrocytic hypertrophy, 

markers of the AD pathology present in elderly DS individuals. 

Moreover, Ts65Dn mice undergo cognitive deterioration beginning 

at 6-8 months of age. All this evidence suggests that Ts65Dn mice 

may provide a unique model which is suitable for following the 

progression of AD. 

While AD develops only in some individuals of the general 

population, it is the invariable hallmark of DS, implying that trisomic 

genes play a paramount role in development of the disease. Indeed, 

recent evidence shows that the presence of an extra copy (due to 

trisomy 21) of the dual-specificity tyrosine-phosphorylated and 

regulated kinase 1A (Dyrk1A) gene results in overexpression of 

Dyrk1A and elevated kinase activity in the DS brain. Dyrk1A 

phosphorylates tau at several sites, and these sites are 

hyperphosphorylated in adult DS brains. Phosphorylation of tau by 

Dyrk1A primes its further phosphorylation by glycogen synthase 

kinase-3beta (GSK-3beta). These findings strongly suggest a novel 

mechanism by which the overexpression of Dyrk1A in DS brains 

causes neurofibrillary degeneration via hyperphosphorylating tau. 

The putative involvement of Dyrk1A in the etiology of AD implies 

that de-regulation of Dyrk1A may be an age-related phenomenon 

that ultimately leads to the pathology. In the Ts65Dn mouse brain, 

an extra copy of the Dyrk1A gene causes increased expression and 

activity of Dyrk1A and results in increased tau phosphorylation.


NEW TREATMENT 

STRATEGIES 





Consequently, Ts65Dn mice may represent a good model for the 

elucidation of the role of Dyrk1A (and other trisomic genes) in the 

development of AD. 

AD is histopathologically characterized by intraneuronal 

neurofibrillary degeneration of the abnormally 

hyperphosphorylated tau and extracellular β-amyloidosis. The 

neurofibrillary degeneration is apparently required for the clinical 

expression of AD, and in related tauopathies it leads to dementia in 

the absence of amyloid plaques. While normal tau promotes 

assembly and stabilizes microtubules, the abnormally 

hyperphosphorylated tau disrupts microtubules. Tau is 

phosphorylated by a number of protein kinases. Glycogen synthase 

kinase-3 (GSK-3) is one of the key kinases required for AD-type 

abnormal hyperphosphorylation of tau, which is believed to be a 

critical event in neurofibrillary tangle formation. Thus, GSK-3 

inhibition represents a very attractive drug target in AD and other 

neurodegenerative disorders. The inhibition of abnormal 

hyperphosphorylation of tau is one of the most promising 

therapeutic targets for the development of disease modifying drugs. 

By exploiting the Ts65Dn mouse it will be possible to establish 

whether treatments interfering with tau phosphorylation 

ameliorate the AD phenotype. More importantly, it will be possible 

to establish whether pharmacotherapy during the early life stages 

that precede the overt manifestations of AD is able to 

prevent/delay the onset of the pathology 

PRIN 2008 - Effect of prenatal and early postnatal treatment with 

fluoxetine on neurogenesis and dendritic maturation in the Ts65Dn 

mouse model for Down's syndrome 

Abstract del progetto Cognitive decline in Down mice; pharmacological treatment to 

prevent cognitive decline by stimulating neurogenesis

Nome Sonia Levi 

Contatti 

+39 0226434755-53 

levi.sonia@hsr.it 



San Raffaele Scientific Insitute/Division of Neuroscience 

Area di interesse identificata Neurodegeneration 



Functional and epidemiological study of neuroferritinopathies: 

implications for the role of iron in neurodegenerative disorders. 

Ente finanziatore Telethon 

Durata progetto Three years 

Abstract del progetto Iron participates in the oxidative damage common to various 

neurodegenerative disorders, such as Alzheimer and Parkinson diseases 

and Hallevorden−Spatz syndrome, and iron accumulations have been 

frequently observed in the areas of the brain that degenerate. Genetic 

abnormalities of some proteins strictly involved in the regulation of iron 

metabolism are associated to neurodegeneration. Among them, 

neuroferritinopathies are rare autosomal dominant disorders associated 

with mutations of the FTL gene that lead to alterations of the C−terminus 

of ferritin L−chain. They are characterized by abnormal iron deposition in 

the basal ganglia of the brain and by the appearance of ferritin 

aggregates. L−ferritin is dedicated to iron storage, and preliminary results 

indicate that one of the mutations alters iron availability in cellular 

models. The project is aimed at studying the structural and functional 

properties of the ferritin mutants, by developing various cellular models, 

including primary neurons and glial cells. By this approach, we expect to 

provide indications on the molecular mechanisms that are responsible for 

both intracellular iron accumulation and protein aggregate formation. In 

parallel, we plan to produce knock−in mice models expressing the 

mutated ferritins, to study the progress of neurodegeneration as well as 

the formation of ferritin aggregates in basal ganglia. Possible therapeutic 

approaches, including the use of RNA interference, will be evaluated on 

both cellular and animal models. Since DNA variations in FTL gene are not 

uncommon, we plan to analyse its mutations, as well as those of other 

iron−related genes, in patients with neurological disorders. This study is 

aimed to elucidate how iron acts as a cofactor in neurodegenerative 

processes.

Nome Tiziana Antonelli 

Contatti ant@unife.it 



Dept. of Clinical and Experimental Medicine 


Area di interesse identificata Mechanisms of Neurodegeneration : I. Animal models. 



Neurochemical and morphological in vivo and in vitro models in 



BioPharmaNet: Laboratory of research and innovation for the Life 

Sciences sector industries (Emilia Romagna, Italy) 


Abstract del progetto In vitro models of neurodegenerative diseases have been developed and 

optimized. Cortical cell cultures, mainly containing GABAergic and 

glutamatergic neurons, and hippocampal cell cultures, especially 

containing glutamatergic neurons, can be used to study the cell 

degeneration following neurotoxic insults. The neuronal damage may be 

evaluated by analysing neurochemical ([ 3 

H]GABA and [ 3 

H]glutamate 

uptake and release, biochemical (MTT assay, LDH levels) and 

morphological (Hoechst 33258 nuclear staining, MAP-2 immunoreactivity) 

parameters. These in vitro experiments, performed in primary neuronal 

cultures, cerebral tissue slices and/or synaptosomes, are complementary 

to the in vivo microdialysis experiments and by combining them it is 

possible to investigate on the efficacy as well as the neurochemical and 

molecular mechanisms of new drugs against neurodegenerative 

pathologies. In vivo microdialysis technique in unilaterally 

6-hydroxydopamine (6-OHDA) lesioned rats allows to understand the 

possible relevance of new pharmacological approaches in the treatment 

of this pathology. The simultaneous evaluation of the release of several 

neurotransmitters such as dopamine, acetylcholine, glutamate and GABA 

in the striatum, globus pallidus and/or other close related brain nuclei of 

the indirect and direct pathways of the basal ganglia, gives the 

opportunity to evaluate the efficacy of new therapeutic and genetic 

strategies in the control of the “motor” pathways both in physiological 

and pathological conditions.


A- Drug Design

Nome Nino Russo 

Contatti nrusso@unical.it 

Istituto/Dipartimento Dipartimento di Chimica, Università della Calabria, via P. Bucci, cubo14c, 

87036 Rende (CS), Italy 


Alzheimer_s disease (AD) is characterized by the deposit of extracellular amyloid plaques, which consist 

predominantly of the aggregated peptide amyloid-b (Ab). The aggregation mechanism is not very well 

understood, but it is proposed that aggregated forms of Ab are neurotoxic because of the 

production of reactive oxygen species (ROS). Copper and other metal ions (e.g. Zn) have been found to 

accumulate in amyloid plaques. Moreover, Cu ions have been proposed to be linked to the aggregation of 

the Ab peptide and/or to the increased generation of ROS. In this context, Cu II coordination to Ab 

becomes a key feature, because it will directly affect the structure of the peptide and, hence, the 

aggregation behaviour. 

The catalytic ROS production of the Cu II ions coordinated by Ab will also be governed by the binding 

mode. Therefore, this system has been extensively studied. However, the unambiguous identification of 

the Cu II ligands has remained difficult and no real consensus has emerged in the literature yet. It 

has been shown that all of the ligands of the high affinity Cu II site are contained in the Ab16 N-terminal 

domain (DAEFRHDSGYEVHHQK); hence, this soluble fragment has been widely used as a model for the CuII 

binding site in the native Ab40/42 peptides. Although a lot of experimental studies performed with a wide 

range of methods, the analysis of the results difficult and impaired the unambiguous identification of the 

Cu II ligands. 

Modern theoretical and computational methods can give a significant contribution to the knowledge of 

the elementary mechanisms at atomistic level on a series of open questions such as: i. unambiguous 

identification of the metal coordination site; ii. the role of the metal coordination in the conformational 

change of the metallated protein; iii. The effect of the environment on the complex formation; iv. Design 

of new and specific drugs for the metal chelation and sequestration. 

For the development of this research program, sophisticated computational protocols as well as 

theoretical methods will be implemented and used. 




Development of computational and theoretical methods based on the 

Density Functional theory and their application on systems with biological 

significance. 

Ente finanziatore Università della Calabria 

Durata progetto Due anni 

Abstract del progetto New algorithms and computational protocols based on the density 

functional theory should be proposed and tested. The applications should 

be essentially devoted to the study of complex biological systems

Nome Carlo Melchiorre 

Contatti e-mail: carlo.melchiorre@unibo.it ph: 051-2099706 

Istituto/Dipartimento Dipartimento di Scienze Farmaceutiche 


Università di Bologna 

Via Belmeloro, 6, 40126 Bologna 

Innovative drug discovery strategies for Alzheimer’s disease 


New treatment strategies 


Alzheimer’s disease (AD) is a major neurodegenerative disease 

affecting a substantial proportion of elderly population and 

imposing a significant burden on public health. For AD there is still 

no cure, and only few symptomatic treatment options are available 

to the 30 million patients worldwide. Driven by the clear unmet 

medical need and a better understanding of the AD biology and 

pathophysiology, several lead candidates have progressed to preclinical 

testing in the last decade. However, to date none has been 

successful in late-stages clinical trials. Currently, AD is best 

characterized as a multidysfunctional condition, where interrelated 

molecular events result in amyloid formation, tau abnormalities, 

amyloid accumulations, and loss of acetylcholine neurotransmission. 

Currently available drugs and most of the drugs under 

development target only one of these mechanisms. This may 

explain why they exhibit limited effectiveness. Therefore, 

therapeutic tools able to confront this complexity are urgently 

needed. Another issue to take into account in AD drug discovery is 

that AD patients are susceptible to a wide range of concomitant 

medical conditions (co-morbidity), including hypertension, vascular 

diseases, diabetes, which can often be associated. There is 

therefore a need for therapeutic tools to be tailored to their specific 

conditions. 

In this respect, we have recently proposed that multitarget ligands, 

i.e. single chemical entities able to modulate simultaneously 

multiple targets, might represent the best pharmacological option 

for both tackling the multifactorial nature of AD and simplifying AD 

patient therapeutic regimen. Furthermore, from a chemical biology 

perspective, we envisaged that rationally designed multitarget 

chemical probes have promise for studying the complex 

mechanisms underlying neurodegeneration.


Principal Investigator: “Neuroreceptors machinery: design and 

synthesis of molecules useful for their characterization and 

modulation and for the treatment of related dysfunctions” 

(20073EWPF9_003;) 

Ente finanziatore Ministero dell’Istruzione, dell’Università e della Ricerca (€ 490,000) 

Durata progetto 22/09/2008-21/09/2010 



The present project regards the design, synthesis and preliminary 

pharmacological evaluation of ligands and modulators of 

neurotransmitter receptor systems. The knowledge of structure and 

functions of neurotransmitter receptor systems is critical to design 

and develop drugs useful to treat the numerous pathologies that 

depend on central and peripheral nervous transmission 

dysfunctions. As a matter of fact, it is well known that severe 

pathologies such as pain, anxiety, mental conditions, and 

neurodegeneration are due to receptor dysfunctions. Some of these 

pathologies have a tremendous social impact as they involve large 

population segments, in particular aged people. 

Principal Investigator: “Development of Innovative Methodologies 

for the Identification and Synthesis of New Bioactive Molecules: 

application to the Alzheimer's disease” (FIRB 2003 RBNE03FH5Y;) € 

Ente finanziatore Ministero dell’Istruzione, dell’Università e della Ricerca (€ 

2,871,000) 



The modern Medicinal Chemistry is nowadays requiring an actual 

multidisciplinary approach and the development of new 

technologies either to accelerate the whole discovery process, or to 

provide better and safer candidate drug. In this context, the goals of 

the present project are both to develop innovative technological 

frameworks for molecular recognition and combinatorial chemistry, 

and to validate the applicability of such techniques through the 

production of new therapeutics in the field of the Alzheimer's 

disease (AD)

Nome Prof. Federico Da Settimo, Prof. Claudia Martini 

Contatti fsettimo@farm.unipi.it, cmartini@farm.unipi.it 

Istituto/Dipartimento Department of Pharmaceutical Science, Department of Psychiatry, 

Neurobiology, Pharmacology, and Biotechnology - University of Pisa 


Development of labelled ligands specifically targeting the Translocator Protein 18 kDa in Alzheimer 

Disease. 

The Translocator Protein (TSPO) is a mitochondrial transmembrane protein involved in a variety of 

biological processes, such as steroidogenesis, apoptosis induction and inflammation, and it is differently 

expressed under physio/pathological conditions, in brain cells (astrocytes and microglia). A dramatic 

increase in TSPO levels occurs in glial cells in response to brain injury and inflammation, so that TSPO can 

be considered as a biomarker of ongoing brain insults. The activation of microglial cells and the 

consequent up-regulation of the TSPO sites have been demonstrated in both normal ageing and in brains 

of patients affected by Alzheimer disease (AD), stroke and multiple sclerosis. Increased TSPO expression 

has been observed In this view, the use of TSPO ligands could efficiently reveal microglia activation, so 

representing a useful tool in brain imaging, both in vivo and in post-mortem analysis. 

Our group has gained experience in the synthesis of heterocyclic compounds acting at the 

mitochondrial translocator protein (TSPO). As well documented by several international publications, over 

the course of several years the research group has also gained experience in the evaluation of the 

biological activity of new synthesised compounds, through high yield methodologies (Primofiore, G., et al. 

J. Med. Chem. 2004, 47, 1852; Chelli, B., et al Chembiochem. 2005, 6, 1082; Taliani, S., et al. J. Med. Chem. 

2007, 50, 404; Da Settimo, F., et al. J. Med. Chem. 2008, 51, 5798; Chelli, B., et al. J. Cell. Biochem. 2008, 

105, 712.) 

The gained experience have generated significant efforts of the group to apply TSPO ligands as 

marker of “active” brain pathology, in line with multiple research groups throughout the world. 

In the present project we aim to design, synthesise and evaluate novel specific TSPO probes 

featuring the indolglyoxylamide scaffold, suitable as useful tools in brain imaging. A number of 

radiolabelled ligands targeting TSPO will be developed as powerful tools to image and measure the 

expression levels of the protein in humans, opening up the opportunity for the discovery of specific PET 

ligands for TSPO. PET studies of TSPO could offer quantitative follow-up of neuroinflammation, an 

advantage for biomarker-type measurement during drug development and early clinical development. 

Preliminarily, studies are in progress with monkeys. 

Moreover, fluorescent ligands, that represent a safer, faster, and less expensive alternative to 

radioligands, will be developed for usage in biomedical research. In particular we will investigate a series 

of fluorescent probes characterized by the presence of a chemoreactive group able to bind the receptor 

protein irreversibly and covalently, in order to overcome the fact that tissue experimental procedures 

using a labelled ligand often cause the alteration of the chemical equilibrium and the subsequent loss of 

the bound fluorescent ligand. The presence of these two devices (a chemoreactive group and a fluorescent 

chromophore) on a single molecule may offer a multiplicity of advantages, both in protein 

purification/characterization, and in protein cell visualization/density determination in both cell cultures 

and tissue slices derived from post-mortem brains. 

Pursuing of these goals will be realized in strict collaborations with the research groups of Prof. 

Silvia Selleri (University of Florence). 



Drug development for AD Diagnosis, Monitoring and Brain imaging


Ente finanziatore University of Pisa 


24 months 


New fluorescent tools targeting the translocator protein (TSPO) as 

diagnostics for cancer and neurodegenerative diseases. 

The proposed research is addressed to develop innovative fluorescent 

probes targeting the 18 kDa Translocator Protein (TSPO) (previously 

known as the peripheral-type benzodiazepine receptor) as promising 

molecular imaging (MI) tools to be widely applied in studying TSPO 

location, turn-over and interaction as well as in diagnosing TSPO related 

pathologies. TSPO is a protein mainly expressed in mitochondria, which is 

involved in cell proliferation, apoptosis, immunomodulation and 

steroidogenesis. TSPO basal density is altered in pathologies in which a 

perturbation of cell survival/death processes has been evidenced such as 

cancer and neurodegenerative disorders and alterations in TSPO 

subcellular distribution have been correlated with the malignancy grade 

of cancer cells. Therefore, TSPO has been suggested as a promising drug 

target as well as a valid diagnostic marker of state and progression of 

such pathologies and has become an interesting target in MI. Currently, 

MI has emerged as an important multidisciplinary area, which involves 

radiology, biology, biochemistry, physics, engineering, medicine and 

synthetic chemistry, essential in providing potent imaging agents. Based 

on our previous experiences, the aim of this research program is to 

design, synthesize and biologically characterize/validate new reversible 

and irreversible fluorescent 2-phenylindol-3-ylglyoxylamide ligands with 

high affinity and selectivity for TSPO. The development of these 

innovative fluorescent TSPO ligands will offer a wide range of advantages 

for their applications in molecular basic studies as well as in diagnosis.

Nome Alberto Cassetta 

Contatti 

Alberto.cassetta@ts.ic.cnr.it - Tel: ++39 040 226881 

Istituto/Dipartimento CNR – Istituto di Cristallografia UOS Trieste (Dip. Progettazione 

Molecolare) 


Structural studies of acetylcholinesterases and their inhibitors: implications for the design of new anti-Alzheimer 

drugs 

Acetylcholine esterase (AChE) plays a crucial role in the regulation of signal transmission, by terminating the action of 

acetylcholine at the cholinergic synapse and muscular junctions. AChE inhibitors are used in the symptomatic 

treatment of pathologies such as Alzheimer's disease. The 3D crystal structures of a number of AChE complexes with 

reversible and irreversible inhibitors have been determined. The structural information accompanied by kinetic 

studies will allow a detailed understanding of the molecular determinants underlying the mechanism of action of this 

enzyme. Furthermore these results are expected to offer a rational framework for the design and synthesis of 

compounds with optimal pharmacokinetic and pharmacodynamic properties. 

Structural studies of neurotrophins (NGF), proneurotrophins(proNGF) and their interactions with their receptors 

TrkA, p75, sortilin) and a panel of neutralizing antibodies(MNAC13, αD11) 

Neurotrophins such as nerve growth factor (NGF) are involved in the development and maintenance of neurons both 

in the central and peripheral nervous system, and have been implicated as potential therapeutic target molecules in 

the Alzheimer's disease. NGF exerts its biological function by binding to TrkA receptor promoting dimerization, 

catalytic auto-phosphorylation of intracellular tyrosine residues and triggering of the signal transduction cascade. 

Until recently, no clear biological role had been attributed to the proneurothrophin (proNGF) peptide, except helping 

in folding and secretion of mature NGF. Moreover, proNGF was found to be the predominant form of NGF in brain 

and to be increased in Alzheimer's disease and to induce p75NTR dependent apoptosis in cultured neurons. The 

specific receptor for proNGF is sortilin. We are investigating the structural and functional bases of the biological 

differences between NGF and proNGF. The monoclonal antibodies MNAC13 and αD11 are potent antagonists that 

prevent the NGF-TrkA interactions in vivo systems. In order to gain deeper insight into the molecular basis of the 

observed high affinity of NGF for TrkA, we are investigating the 3D structures of both neutralizing antibodies and of 

the respective complexes. Detailed structural information on their antigenic recognition is expected to aid in the 

development of analogs as antagonists or agonists of neurotrophins that may have greater affinity or specificity for 

further experimental and therapeutic applications. 

Structural studies of Hydroxysteroid Dehydrogenases and their interactions with phytoestrogens offlavonoid 

nature. 

17β-Hydroxysteroid dehydrogenases (17β-HSDs) are enzymes which play a crucial role in steroid hormones action by 

acting on the final steps of estrogen or androgen biosynthesis. Given their ability to interconvert a hormonally 

inactive (or less active) steroid to its active form, these enzymes operate a pre-receptorial regulation of steroid 

action. Several distinct 17β-HSDs isozymes can be found in human, principally distinct by their tissue type and cellular 

localization, homology sequence and metabolic pathway. In humans different 17β-HSDs have been linked to several 

pathological forms: prostate and breast cancer (17β-HSD type 1) and Alzheimer’s disease (17β-HSD type 10) among 

others. We have recently undertaken a structural study on a fungal 17β-HSD which share a remarkable structural 

similarity with both type 1 and 10 of human 17β-HSD. Our studies were aimed to investigate the use of the fungal 

enzyme as possible model for the understanding at the molecular level the interaction mechanism of 17β-HSDs with 

specific and less-specific inhibitors. In particular we are investigating the inhibitory action of phytoestrogens,

principally of flavonoid nature, towards 17β-HSD type 1, which have been demonstrated to be inactivated at the 

μmol level by certain flavonoids and coumestans. Moreover, given the lack of biochemical and structural data about 

the effects of flavonoids on type 10 17β-HSD, we intend to further investigate this subject by a 

biochemical/biophysical approach combined with crystallographic methods by studying the inhibition properties of 

the above compounds toward the human 17β-HSD homologue. Because both mutations of the gene encoding this 

enzyme and its overexpression give raise to a distinct type of mental retardation (17--β-Hydroxysteroid 

Dehydrogenase X Deficiency; OMIM #300438) and it has been shown to be a molecular link between mitochondrial 

dysfunction and Alzheimer’s disease by direct binding of the β-amyloid peptide, our approach will provide a valuable 

understanding of the role played by this enzyme in the occurrence of these neuropathies. 

LUSTBADER J, CIRILLI M., LIN C., XU H.W., CASPERSEN C., WANG N., TAKUMA K., CHEN X., CHANEY M. TRINCHESE 

F., LIU S., KUPPUSAMY P., ZEWIER Z., ARANCIO O., STERN D., YAN S.D., WU H.: “ABAD Directly Links Aβ to 

Mitochondrial Toxicity in Alzheimer’s Disease”. Science (2004) 304, 448-52 

Paoletti F, Covaceuszach S, Konarev PV, Gonfloni S, Malerba F, Schwarz, E,Svergun DI, Cattaneo A, Lamba D. “Intrinsic 

structural disorder of mouse proNGF”. Proteins. 2009 Jun;75(4):990-1009. 

Covaceuszach S, Cassetta A, Konarev PV, Gonfloni S, Rudolph R, Svergun DI, Lamba D, Cattaneo A. “Dissecting NGF 

interactions with TrkA and p75 receptors by structural and functional studies of an anti-NGF neutralizing antibody”. 

J Mol Biol. 2008 Sep 12;381(4):881-96. 

Bartolucci C, Haller LA, Jordis U, Fels G, Lamba D. “Probing Torpedo californica acetylcholinesterase catalytic gorge 

with two novel bis-functional galanthamine derivatives”. J Med Chem. 2010 Jan 28;53(2):745-51. 

Cattaneo, S. Covaceuszach, D. Lamba: “Method for the humanization of antibodies and 

humanized 

antibodies thereby obtained”. Patent WO/2005/061540 


identificata 

Structural Neurobiology 

Finanziamenti ricevuti Not Applicable 






B-Drug screening

Nome Laura Calzà 

Contatti 


Tel. +39 051 2097947 

Cell. +39 335 310979 

Istituto/Dipartimento DIMORFIPA 



Development of competitive 

animal models for AD 

Basic research 


GLP grade animal facility and behavioural testing for cognitive 

performance in AD animal models using video tracking and 

computer analysis; screening of new molecules after acute 

administration 

Brain metabolism of thyroid hormone: a risk factor for AD? We are 

interested in exploring possible connections between thyroid 

hormone content, receptor and activating enzyme expression, APP 

metabolism and Ab toxicity in the brain in normal and AD mice. 

Recent advances in knowledge on thyroid hormone brain 

metabolism and molecular biology of nuclear and membrane 

thyroid hormone receptors and transporters offer new possible 

targets to explore this topic.

Nome Iliana Ferrero Fortunati 

Contatti 

iferrero@unipr.it 

Phone: +39-0521905600 

Fax: +39-0521905604 

Mobile : +39-3480030063 

Istituto/Dipartimento Dipartimento di Genetica, Biologia dei microrganismi, Antropologia, 

Evoluzione – Università degli Studi di Parma 

Parco Area delle Scienze 11/A - Campus 

43100 Parma 


“Yeast model to seek for molecules which can reduce mitochondrial mutability” 

Mitochondrial dysfunction is a prominent feature of Alzheimer’s Disease (AD) and there is growing body 

of evidence that mitochondrial dysfunction plays a crucial role in the pathogenesis or influences the risk of 

AD. 

The Yeast Genetics Group of University of Parma, coordinated by of Prof. Iliana Ferrero, has a long lasting 

experience in the use of Saccharomyces cerevisiae as a model system to study mitochondrial mutability to 

address the question of whether there is a molecular relationship between the pathology and the specific 

mutation and to study the pathogenic mechanisms of human disease associated to mitochondrial DNA 

mutations. The yeast group has a forty years experience on yeast genetics and a ten years experience in 

the use of yeast model of mitochondrial diseases (in collaboration with Dr. Massimo Zeviani, Unit of 

Molecular Neurogenetics, IRCCS Foundation Neurological Institute FC. Besta, and with Prof. Giacomo 

Comi, Department of Neurological Sciences and Centre of Excellence on Neurodegenerative Diseases, 

University of Milan). We have created yeast models of several mitochondrial pathologies due to mutations 

in a relevant number of genes involved in mitochondrial DNA stability. Moreover, we have already used 

the S. cerevisiae model system to study the effect of molecules on mitochondrial DNA mutability and we 

have identified molecules able to reduce mitochondrial mutability. 

Specific aims 

The main scope of our proposal is the use of S. cerevisiae models already existing in our collection and, 

when necessary, create new yeast models to test a pharmacological approach to therapy. To this aim we 

propose to subject yeast model to chemicals (chemical library) screening to prospect new therapeutic 

drugs able to reduce either extended or point mutability or both. 

Publications of Iliana Ferrero on yeast model of mitochondrial disease 

Dallabona, C., Marsano, R.M., Arzuffi, P., Ghezzi, D., Mancini, P., Zeviani, M., Ferrero, I., Donnini, C. (2010). 

Sym1, the yeast ortholog of the MPV17 human disease protein, is a stress-induced bioenergetic and 

morphogenetic mitochondrial modulator. Hum Mol Genet 2010 Jan 15. [Epub ahead of print] 

Ghezzi D, Goffrini P, Uziel, G, Horvath R, Klopstock T, Lochmaller H, D'Adamo P, Gasparini P, Strom TM, 

Prokisch H, Invernizzi F, Ferrero I, Zeviani M (2009). SDHAF1, encoding a LYR complex-II specific assembly 

factor, is mutated in SDH-defective infantile leukoencephalopathy. Nature Genetics, 41, 654-656. 

Di Fonzo, A., Ronchi, D., Lodi, T., Fassone, E., Tigano, M., Lamperti, C., Corti, S., Bordoni, A., Fortunato, F., 

Nizzardo, M., Napoli, L., Donadoni, C., Salani, S., Saladino, F., Moggio, M., Bresolin, N., Ferrero, I., Comi, 

G.P. (2009). The mitochondrial disulfide relay system protein GFER is mutated in autosomal-recessive 

myopathy with cataract and combined respiratory-chain deficiency. Am J Hum Genet 84 594-604. 

Ghezzi, D., Goffrini, P., Uziel, G., Horvath, R., Klopstock, T., Lochmaller, H., D'Adamo, P., Gasparini, P., 

Strom, T.M., Prokisch, H., Invernizzi, F., Ferrero, I., Zeviani, M. (2009). SDHAF1, encoding a LYR complex-II 

specific assembly factor, is mutated in SDH-defective infantile leukoencephalopathy. Nature Genetics,

41, 654-656 

Goffrini, P., Ercolino, T., Panizza, E., GiachÃ¨, V., Cavone, L., Chiarugi, A., Dima, V., Ferrero, I., Mannelli, M. 

(2009). Functional study in a yeast model of a novel succinate-dehydrogenase subunit B gene germline 

missense mutation (C191Y) diagnosed in a patient affected by a glomus tumor. Hum Mol Genet 18, 

1860-1868. 

Spinazzola, A., Invernizzi, F., Carrara, F., Lamantea, E., Donati, A., Dirocco, M., Giordano, I., Meznaric- 

Petrusa, M., Baruffini, E., Ferrero, I., Zeviani, M. (2009). Clinical and molecular features of mitochondrial 

DNA depletion syndromes. J Inherit Metab Dis 32 143-158. 

Galassi, G., Lamantea, E., Invernizzi, F., Tavani, F., Pisano, I., Ferrero, I., Palmieri, L., Zeviani, M. (2008). 

Additive effects of POLG1 and ANT1 mutations in a complex encephalomyopathy. Neuromuscul Disord 

18 465-70. 

Massa V, Fernandez-Vizarra E, Alshahwan S, Bakhsh E, Goffrini P, Ferrero I, Mereghetti P, D'Adamo P, 

Gasparini P, Zeviani M. (2008). Severe Infantile Encephalomyopathy Caused by a Mutation in COX6B1, a 

Nucleus-Encoded Subunit of Cytochrome C Oxidase. Am J Hum Genet. 82 1281-1289. 

Massa, V., Fernandez-Vizarra, E., Alshahwan, S., Bakhsh, E., Goffrini, P., Ferrero, I., Mereghetti, P., 

D'Adamo, P., Gasparini, P., Zeviani, M. (2008). Severe infantile encephalomyopathy caused by a 

mutation in COX6B1, a nucleus-encoded subunit of cytochrome c oxidase. Am J Hum Genet 82 1281- 

1289. 

Barberio, C., Bianchi, L., Pinzauti, F., Lodi, T., Ferrero, I., Polsinelli, M., Casalone, E. (2007). Induction and 

characterization of morphologic mutants in a natural Saccharomyces cerevisiae strain. Can J Microbiol 53 

223-230. 

Baruffini, E., Ferrero, I., Foury, F. (2007). Mitochondrial DNA defects in Saccharomyces cerevisiae caused 

by functional interactions between DNA polymerase gamma mutations associated with disease in 

human. Biochim Biophys Acta 1772 1225-35. 

Fernandez-Vizarra E, Bugiani M, Goffrini P, Carrara F, Farina L, Procopio E, Donati A, Uziel G, Ferrero I, 

Zeviani M. (2007). Impaired complex III assembly associated with BCS1L gene mutations in isolated 

mitochondrial encephalopathy. Hum Mol Genet. 16 1241-1252. 

Valente, L., Tiranti, V., Marsano, R.M., Malfatti, E., Fernandez-Vizarra, E., Donnini, C., Mereghetti, P., De 

Gioia, L., Burlina, A., Castellan, C., Comi, G.P., Savasta, S., Ferrero, I., Zeviani, M. (2007). Infantile 

encephalopathy and defective mitochondrial DNA translation in patients with mutations of 

mitochondrial elongation factors EFG1 and EFTu. Am J Hum Genet 80 44-58. 

Baruffini, E., Lodi, T., Dallabona, C., Puglisi, A., Zeviani, M., Ferrero, I. (2006). Genetic and chemical rescue 

of the Saccharomyces cerevisiae phenotype induced by mitochondrial DNA polymerase mutations 

associated with progressive external ophthalmoplegia in humans. Hum Mol Genet 15 2846-55. 

Spinazzola, A., Viscomi, C., Fernandez-Vizarra, E., Carrara, F., D'Adamo, P., Calvo, S., Marsano, R.M., 

Donnini, C., Weiher, H., Strisciuglio, P., Parini, R., Sarzi, E., Chan, A., DiMauro, S., RÃ¶tig, A., Gasparini, P., 

Ferrero, I., Mootha, V.K., Tiranti, V., Zeviani, M. (2006). MPV17 encodes an inner mitochondrial 

membrane protein and is mutated in infantile hepatic mitochondrial DNA depletion. Nat Genet 38 570-5. 

Fontanesi, F., Viola, A.M., Ferrero, I. (2006). Heterologous complementation of the Klaac null mutation of 

Kluyveromyces lactis by the Saccharomyces cerevisiae AAC3 gene encoding the ADP/ATP carrier. FEMS 

Yeast Res 6 414-20. 

Lodi, T., Bove, C., Fontanesi, F., Viola, A.M., Ferrero, I. (2006). Mutation D104G in ANT1 gene: 

Complementation study in Saccharomyces cerevisiae as a model system. Biochem Biophys Res Commun

341 810-5 

Palmieri L, Alberio S, Pisano I, Lodi T, Meznaric-Petrusa M, Zidar J, Santoro A, Scarcia P, Fontanesi F, 

Lamantea E, Ferrero I, Zeviani M (2005). Complete loss-of-function of the heart/muscle-specific adenine 

nucleotide translocator is associated with mitochondrial myopathy and cardiomyopathy. Hum Mol Genet 

14 3079-88. 

Fontanesi F, Palmieri L, Scarcia P, Lodi T, Donnini C, Limongelli A, Tiranti V, Zeviani M, Ferrero I, Viola AM 

(2004). Mutations in AAC2, equivalent to human adPEO-associated ANT1 mutations, lead to defective 

oxidative phosphorylation in Saccharomyces cerevisiae and affect mitochondrial DNA stability. Hum Mol 

Genet 13 923-34. 




Yeast model for human health: Neurodegenerative diseases; 


“Identification and characterization of nuclear genes responsible for human 

mitochondrial disorders” Project coordinated by Massimo Zeviani PhD, Unit of 

Molecular Neurogenetics, IRCCS Foundation Neurological Institute FC. Besta. 

Ente finanziatore Telethon Italy – Tree years project – 2008-2010 



Albeit still insufficiently acknowledged, mitochondrial disease encompasses a 

substantial fraction of human inherited disorders. The inclusive term 

"mitochondrial medicine" has recently been coined to account for the broad 

functional implications of mitochondria in normal and disease conditions. 

Mitochondrial disorders are very heterogeneous, can affect any organ, at any 

age, and responsible genes are still missing in most cases. These features make 

the diagnosis of mitochondrial disease difficult, and in many instances cause 

these disorders to be overlooked. Effective therapy is missing as well. 

Nevertheless, great progress has been made in this field in the past decade, 

thanks to the discovery of new genes relevant to disease, and the investigation 

of the physiopathology of these disorders through a multidisciplinary approach. 

In recent years we have been finding several new disease genes, which are 

responsible for well−defined mitochondrial syndromes. However, the function of 

these genes is still completely or largely unknown. 

We shall also create disease models user the friendly systems yeast recombinant 

strains. The study of yeast model will help us understand the physiopathology of 

the disorders caused by the defective genes, gain insight on their function and 

related metabolic pathways, and devise sensible and specific therapeutic 

strategies.

Nome Luca Ferraro 

Contatti 

frl@unife.it 

Istituto/Dipartimento Dept. of Clinical and Experimental Medicine 




identificata 






Pharmacology: B. Drug screening . 

Innovative technologies for the therapy of the neurodegenerative 

diseases 

NeuroBioTech: Innovative technologies for the therapy of the 

neurodegenerative diseases (Emilia Romagna, Italy) 

2009-2012 

The aim of NeuroBioTech is the development of a precompetitive 

research activity to find new therapeutic strategies for the treatment 

of Alzheimer disease (AD) and other correlated neurodegenerative 

pathologies. The experimental activity of our research unit is aimed 

to develop and optimize in vivo models (microdialysis, behavioural 

tests) and ex vivo models (cerebral tissue), which will be applied in 

preclinical testing activity for the screening of molecules with anti- 

Alzheimer activity and for the identification of new therapeutic 

targets in AD. The eventual alterations of endogenous 

neurotransmitter levels (glutamate, GABA, noradrenaline, 

dopamine) will be evaluated in the above reported in vivo and ex 

vivo experimental models. In particular, the eventual modifications 

of neurochemical signal will be analysed in mice with mutations of 

APP and/or PS and/or tau. Furthermore, it will be studied if the 

CHF5074 (new modulator of gamma secretases) administration will 

be able to correct these neurochemical alterations. Then, the effect 

of CHF5074 on the APP metabolism and on APP molecular 

interactions will be studied.


C- Drug Delivery


Contact person: Prof. MA Vandelli, Dr. G Tosi (gtosi@unimore.it) Università di Modena e Reggio 

Date: 14-01-2009 

I) Strategic Issues (Justification of the importance of the issue) 

NANOTECHNOLOGY FOR HEALTH: THE APPLICATION IN THE BRAIN DISEASES 

The application of nanotechnology to health raises high expectations for a more efficient and affordable 

healthcare and has the potential of delivering promising solutions to many illnesses. Even if several areas of 

medical care could benefit from the advantages that nanotechnology can offer, a selective CNS drug 

delivery and targeting could improve the therapy of brain diseases which have a tremendous negative 

impact not only on the patient himself but also on the whole society and linked social and insurance 

systems. 

The project could be also the answer to the fragmentation in nanomedicine research in Europe establishing 

a clear strategic vision in the area. 

In order for CNS drugs to penetrate to the brain tissue, they must pass through the blood brain barrier 

(BBB). Many of the compounds that otherwise would be effective in treating CNS diseases are excluded 

from reaching a sufficient concentration in the brain tissue and producing the desired therapeutic effect. It 

has been estimated that only 2% of the possible CNS therapeutic compounds can cross the BBB. 

Thus, drug delivery to the brain is a challenge, because this tissue benefits from a very efficient protective 

barrier. Even if several disorders and diseases that affect the brain can lead to some loss of BBB integrity, 

this barrier represents an insurmountable obstacle for many drugs able to treat pathologies in which the 

BBB integrity is preserved. 

The same mechanisms that protect the brain from foreign substances also restrict the entry of many 

potentially therapeutic agents. The blood brain barrier is the major barrier to the passage of active 

molecules from the blood compartment to the brain. It is located at the level of the brain capillaries, where 

there is a convergence of different cell types: endothelial cells, pericytes, astrocytes and microglias. The 

brain microvessel endothelial cells that form the BBB, display important morphological characteristics such 

as the presence of tight junctions between the cells, the absence of fenestrations and a diminished 

pinocytic activity, that together help to restrict the passage of compounds from the blood into the 

extracellular environment of the brain. 

A promising strategy for the delivery of therapeutics to CNS could be to associate drugs without any 

modification (prodrugs, for example) to colloidal carriers. These vehicles could deliver numerous drug 

molecules at specific sites by coupling ligands to the surface of the colloids which can be administered 

intravenously for chronic treatment. Moreover, owing to their poor stability in biological fluids, rapid 

enzymatic degradation, unfavourable pharmacokinetic properties and lack of diffusion towards the CNS, 

biotech drugs (peptides, proteins, genes and antisense drugs) may be advantageously formulated in brain 

targeted protective nanocontainers.

Colloidal drug carriers include micelles, emulsions, liposomes and nanoparticles (nanospheres and 

nanocapsules). So far, only liposomes and nanoparticles have been largely exploited for brain drug delivery. 

Recently, the first liposomal preparation of a drug reached the market: it is a formulation of the antitumor 

drug Doxorubicin loaded into PEGyilated liposomes for the i.v. treatment of brain tumors (commercial 

formulation Caelyx). In contrast to liposomes, and despite the abudance of experimental works and 

achievements in the field of polymeric nanoparticle (Np) technology that show the ability of properly 

functionalized nanoparticles to cross the BBB, no nanoparticle-based drug formulations have been 

marketed so far. Drugs encapsulated into poly(butylcyanoacrylate) (PBCA) Np covered by the surface active 

agent polysorbate 80 (Doxorubicin. Loperamide, Dalargin, the anticonvulsivant MRZ 2/576), on the surface 

of HSA Np (the model drug loperamide), unable to cross BBB when administered alone, were able to exert 

an appropriate pharmacological effect. Despite interesting results, PBCA Np have limitations that may 

preclude or at least limit their potential clinical applications; its use for humans is at the moment not 

approved by FDA. Thus there is a need for the development of nanoparticles based for example on the 

polymer poly(D,L-lactide-co-glycolide (PLGA), which is approved by FDA for human use, and very promising 

results were found in the literature. 

So far, only a low amount of injected nanoparticle dose is able to reach the brain (in the order of 1%); thus 

active targeting of the BBB represents a promising non invasive strategy for improving drug delivery to 

brain. In fact, the use of ligands specific for receptors present in high percentage on the BBB could 

represent the way to realize a selective delivery of nanoparticles, and thus of the embedded drugs, to CNS. 

With this aim, positive results were obtained when nanocarriers have been conjugated to ligands (OX26 

mAb; transferrin, apolipoproteins or surface-active agents (polysorbate-80) able to bind some of the 

apolipoproteins present in blood that confer the ability to cross the BBB, insulin, folic acid) for active 

targeting, but other ligands have to be discovered and tested for their ability to carry into CNS drug-loaded 

nanoparticles. 

The application of nanotechnology to health raises high expectations for a more efficient and affordable 

healthcare and has the potential of delivering promising solutions to many illnesses. Even if several areas of 

medical care could benefit from the advantages that nanotechnology can offer, a selective CNS drug 

delivery and targeting could improve the therapy of brain diseases which have a tremendous negative 

impact not only on the patient himself but also on the whole society and linked social and insurance 

systems. 

The project could be also the answer to the fragmentation in nanomedicine research in Europe establishing 

a clear strategic vision in the area. 


Health; Neurodegenerative Diseases 

Innovative strategies for the treatment of brain pathologies 

Nanotechnology applied to Health, Nanomedicine 

III) Impact 

The pharmaceutical treatment of central nervous system (CNS) disorders is the second largest area of 

therapy, following cardiovascular disease. U.S. sales for CNS drugs exceeded $ 53 billion in 2002 to 2003.

CNS disorders are five of the top 10 causes of disability. Stroke is the third leading cause of death and costs 

the economy $ 40 billion annually. Fifteen million people suffer from Alzheimer’s disease, which is the 

second most expensive disease to the economy at $ 100 billion annually. Many brain diseases do not have 

satisfactory treatments. Clearly, CNS disorders are an important current and future priority for the 

pharmaceutical industry. 

Moreover, the development of new drug molecules is expensive and time-consuming. The average cost and 

time for the development of a new chemical entity are much higher (approximately $ 500 million and 10-12 

years, respectively) than those required to develop a novel drug delivery system. Hence, there is a need for 

evolving an existing drug molecule from a conventional form to a novel delivery system that can 

significantly improve its performance in terms of patient compliance, safety and efficacy by targeting to the 

specific site. 

Partner involved: 

• Italy (ACADEMY) (PROPONENT) (Prof. Vandelli, Department of Pharmaceutical Sciences, Dean of 

Faculty of Pharmacy, University of Modena and Reggio Emilia, T.e.Far.T.I.) 

o Nanotechnology, nanoparticles, liposome, surface modification, derivatization of biodegradable 

polymers, in vivo experience on animals 

o In vivo pharmacology expertise on CNS drug efficacy 

o Preparation and loading of engineered Np with selected drugs or active substance 

• Switzerland (ACADEMY) 

o Nanotechnology, Coupling methodology with preformed Np , in vitro tests on BBB culture 

• England (ACADEMY) 

o Nanomedicines based on novel nanomaterials of synthetic and biological nature 

• France A (SME) 

o In vivo models of CNS disease (Parkinson’s Disease, Brain Tumor Glioblastoma, Stroke models) 

o In vivo toxicity 

o PK, Biodistribution studies 

• France B (ACADEMY) 

o in vitro BBB (astrocyte/endothelial cell co-cultures), models of pathological BBB (inflamed BBB) 

o HTS platform for the development of peptide-vectors for Np, drug and imaging agent targeting to 

the CNS across the BBB via receptor mediated trancytosis (RMT). 

• France C (ACADEMY) 

o Gene therapy 

� Delivery of siRNA targeted against the prion protein (in vitro and in vivo) 

� Delivery of growth factors (bdnf etc..). 

� in vivo and in vitro experimental models. 

• France D (SME) 

o Imaging techniques 

o Optical Imaging - Scintigraphy - Autoradiography 

• Portugal (SME) 

o Synthesis of peptides and antibodies 

o Gene therapy 

• Czech Republic (ACADEMY) 

o Toxicity of Nanoparticles 

o Interaction with drug metabolising enzymes 

• Denmark (ACADEMY) 

o In vivo experiments on olivocerebellar degeneration and Purkinje cell death and excitotoxic model 

of epileptic seizures

o Novel peptides for the engineering of the Np (15 amino acids (aa) derived from the (61 aa) 

endogenous protein metallothionein (MT). 

• Israel (SME) 

o Imaging 

o Amyotrophic Lateral Sclerosis

Nome Angelo Montenero 

Contatti 

angelo.montenero@unipr.it 

0521905553 

Istituto/Dipartimento Dipartimento di Chimica G.I.A.F. – Università degli Studi di Parma 


Area di interesse identificata Our proposal in this study is the use of sol–gel technique to synthesize 

mesoporous and nanostructured reservoirs which could be implanted 

giving a sustained release of the drug molecules. The devices will be 

constituted by a matrix that contains the drug dispersed or suspended in 

a fine state. The mechanism, as well as the kinetics and rate of drug 

release will depend upon the physicochemical properties of the drug, the 

materials, and the device construction. They will be basically made of 

oxides derived from Titanium, Silicon (or other metals) alkoxides. The 

inner structure of the inorganic matrix will be tailored (playing with the 

conditions of reaction and the relative amounts of the alkoxides, for 

example) to the specific peculiarity of the substance and to the length of 

the administration period. 



The use of alkyl substituted alkoxides together with non-substituted 

alkoxides could be useful to modify the internal network structure, which 

depends on the respective rates of hydrolysis and condensation, and 

therefore on steric hindrance, inductive effects (of alkyl substituents) and 

functionality (number of alkoxy groups) on the central atom as well as on 

catalysis and solvent. 

In cooperation with some European and Mexican research centers we are 

working with sol-gel devices used to release anticonvulsant drugs, such as 

phenytoin and valproic acid; similar reservoirs could be employed with 

drugs for Alzheimer’s treatment. 

Basic Research in Advanced Innovative Nanomaterials: Applications to 

the Solution of Brain Disorders 

(Identifier: FP7-NMP-2007-LARGE-1) 

Ente finanziatore Università Autonoma Metropolitana di Città del Messico - CEE 

Durata progetto Two years 

Abstract del progetto The aim of the group is to study the treatment of three serious 

neurological disorders: Epilepsy, Parkinson’s and Alzheimer’s, using an 

interbrain nanodevice to deliver the drug in front of damaged membrane 

neurons, in order to obtain a potential effect and avoid the 

decomposition of the molecule. 

Nanostructured materials biocompatible with brain tissue will be 

developed. The solids will include ordered silica (SiO2) and ordered titania 

(TiO2), nanostructured manganese oxides (MnOx) cryptomelane type. Also 

the sol-gel nanomaterials: SiO2, TiO2, ZrO2, HfO2, hybrid nanostructured 

materials and monoliths will be prepared using a variety of alkoxides and

y thoughtfully varying synthesis conditions. All the inert matrices will be 

designed using soft chemistry and will require functionalization in order 

to be biocompatible with the surrounding neural membranes. The drugs 

to be encapsulated within the devices include for Epilepsy, valproic acid, 

sodium phenytoine, leviracetam and topiramate; for Parkinson, L-Dopa, 

and dopamine. For Alzheimer the aim is to quantify the change in 

concentration of Filaments tangles Helicoidally (PHFs) and 

glyceraldehyde-3-phosphate dehydrogenase (GAPDH), in presence of 

selective serotonine adsorbent nanomaterial.

Nome MASSERINI MASSIMO 

Contatti 

0264488203 

Massimo.masserini@unimib.it 

Istituto/Dipartimento Dip. Medicina Sperimentale 

Via Cadore 48 

20052 Monza 

Università Milano Bicocca 


New nanodevices overcoming the blood-brain barrier, for the therapy of Alzheimer Disease 

Nanodevices offer an attractive mean of performing therapy and diagnosis with their high potential for 

multi-task functionalization. Theoretically, multi-task functionalization may confer on nanodevices a wide 

array of properties, such as stealth characteristics once they enter the bloodstream (ability to avoid the 

reticulo-endothelial system, RES), ability to cross the blood-brain barrier (BBB) and, of course, ability to 

reach a target. In addition, since it is possible to confer on them features of biocompatibility, lack of 

toxicity and immunogenicity, biodegradability, simple preparation and high physical stability, the 

nanotechnology -based systems are intelligent tools for diagnostics, prognostics, and controlled and 

sustained delivery of therapeutic agents to specific targets . 

However, the BBB represents a formidable challenge to the delivery of therapeutic agents to the central 

nervous system, and in particular for the treatment of Alzheimer disease, due to its complex, high 

selectivity, to the passage of desired molecules into the brain parenchyma. This fact represents an 

handicap also for testing new and future drugs. These issues has not yet been solved with the existing 

nanotechnologies, and need strong investments in the future years. For this, completely new approaches 

are necessary. 

The proposal is to realize new nano-devices, for overcoming the BBB, for the therapy of the Alzheimer 

disease. For this purpose, new approaches should be sought,, such as the possibility of nanodevices to 

carry molecular factories able to perform small tasks; example are the ability to enzymatically modify the 

architecture of the neuronal membrane modifying the activity of resident molecules involved in the 

mechanisms of the disease. 

Area di interesse identificata Nanomedicine 


2.930.387,00€ 

Titolo progetto Nanoparticles for therapy and diagnosis of Alazheimer Disease 

Ente finanziatore EC 

Durata progetto 5 y (2008-2013) 

Abstract del progetto A hallmark of the disease in the AD brain is extracellular aggregates 

(plaques) of the cytotoxic peptide β-amyloid (Aβ). This project intends to 

use nanoparticles (NPs) for therapy and diagnosis, singly or combined 

(theranostics), focusing on brain Aβ as the target. Brain and blood Aβ are 

in equilibrium across the blood-brain barrier (BBB), so the project also 

considers blood Aβ as a target. 

Different NPs will be multiple-functionalized with: i) molecules interacting 

with Aβ, ii) molecules stimulating BBB crossing, ii) PET or MRI contrast 

agents. Artificial and cellular models will be used ; the efficacy of NPs will 

eventually be evaluated in rodent models of AD. Different routes of 

administration (i.v., oral, nasal) or different protocols (two-step, NP

cocktails, aerosols) will be employed to boost NP brain delivery. The 

prediction is that NPs will range, detect, disaggregate, and remove brain 

Aβ. Besides this action, NPs will interact with blood Aβ, drawing out the 

excess of brain peptide by a “sink” effect.


D-Novel Drugs

Nome 

Contatti 

Lamberto Maffei 

maffei@in.cnr.it 

Istituto/Dipartimento Istituto di Neuroscienze del CNR, Pisa 


In human subjects, building on the results obtained in subjects with Mild Cognitive Impairment (MCI) or in 

the very initial stages of Alzheimer’s Disease (AD) with the ongoing project “Train the Brain”, we plan to: 

extend the intervention to a second group of subjects with MCI 

assess the efficacy of the training employed in “Train the Brain” in subjects in more advanced stages of AD 

determine the extent of the training beneficial effects by means of extensive follow up of the subjects 

included in “Train the Brain” 

determine the most effective components of the training 

In murine AD models, we plan to: 

characterize the cellular and molecular mechanisms underlying the action of Intranasal administration (IA) 

of Neurotrophins and of Environmental Enrichment (EE) in preventing the onset or in ameliorating the 

progression of the disease. 

develop new non invasive strategies aimed at preventing or rescuing cognitive deficits and at preventing 

the onset or reducing the progression of the neurodegeneration 

We shall also evaluate whether exposure to an “impoverished” environment accelerates the cognitive 

decline in animal models of AD. 

We have already exploited IA of NGF (De Rosa et al., 2005) to rescue visual memory deficits in a mouse 

model of NGF deprivation which exhibits all hallmarks of human AD and we have recently shown in the 

same model that EE prevents the onset of memory deficits and ameliorates AD hallmarks (Berardi et al., 

2007). We have also recently shown that EE and fluoxetine treatment enhances adult visual cortical 

plasticity to the point of promoting recovery from defective visual development (Sale et al., 2007; Maya 

Vetencourt et al., 2008; Sale et al., 2009); cortical plasticity enhancement seems highly effective in 

rescuing cognitive deficits in models of neurodegeneration even in advanced stages of neuronal loss 

(Fischer et al, 2007). 





- Translational research 

Train the brain: clinical and experimental 

study of the efficacy of cognitive training and 

physical exercise in dementia




Fondazione Cassa di Risparmio di Pisa 

May 2009-May 2012 

Age-dependent cognitive decline is destined to become a highly 

impacting problem at the clinical, economical and assistance level. Age, 

indeed, is the major risk factor for dementia. In Italy, there are around 

700.000 patients diagnosed with dementia and around 100.000 new 

cases every year. 

Among the pathologies which may lead to dementia, Alzheimer's Disease 

(AD) and vascular dementia (VaD) are by far the most frequent. 

At present, there are no effective therapeutic strategies for AD or VaD, 

which are still pathologies orphan of treatment. It is therefore more and 

more apparent the need of testing, validating and implementing new 

strategies aimed at preventing and/or slowing down cognitive decline, 

starting from the early stages of the disease. We propose to assess the 

efficacy of a combination of physical exercise and cognitive training in 

slowing down or arresting the progression of symptoms in subjects at risk 

of or at the very initial stages of AD and VaD. The scientific rationale for 

this proposal is described below. 

a): several studies in humans have demonstrated that the exposure to a 

cognitively and socially stimulating environment and physical exercise 

have beneficial effects on brain functioning, especially in the elderly, and 

reduce the risk of developing dementia (e.g. Laurin et. al, 2001; Fratiglioni 

et al., 2004; Podewils et al., 2005; Marx, 2005; Kramer and Erickson, 

2007). The estimated reduction in the risk of developing dementia varies 

from study to study but generally ranges between 20 and 50%. 

b) In parallel, a large number of studies performed in animal models has 

demonstrated that physical exercise and expossure to a cognitive and 

socially stimulating environment (a combination known as “enriched 

environment”, EE) ameliorates cognitive performance, slow down the 

decline associated with aging, are neuroprotective, and enhance neural 

plasticity (Cotman and Berchtoldt, 2002; Nithianatharajah and Hannan, 

2006). EE ameliorates cognitive deficits in animal models of AD (Adlard et 

al., 2005; Jankowsky et al., 2005) and is also capable to revert cognitive 

deficits when they are already well evident. These results show the EE 

potential as non pharmacological therapeutic strategy not only to prevent 

the onset of cognitive deficits but also to rescue them. 

Studies on the efficacy of a combined physical and cognitive intervention 

in subjects with dementia are very few and present methodological limits. 

AIMS 

Primary: To assess the efficacy of protocols of physical exercise and 

cognitive stimulation on the progression of the disease in subjects at risk 

of, or with AD and VaD in the initial stages of the disease, identified 

through an advanced battery of diagnostic tests. 

Secondary: To develop a nonpharmacological preventive/therapeutic 

strategy easily applicable to humans and exploitable by the National and 

Regional Health Service and to study the underlying cellular and 

molecular mechanism of action in animal models.

Nome Prof. Aldo Andreani 

Contatti 

Prof. Aldo Roda 

aldo.andreani@unibo.it Tel. +39 051 2099714 Fax: +39 051 

2099734 

aldo.roda@unibo.it Tel. +39 051 343398 Fax: +39 051 

343398 

Istituto/Dipartimento Dipartimento di Scienze Farmaceutiche - Università di Bologna 


Via Belmeloro 6 40126 Bologna 

Synthesis and screening of new AChE/BuChE inhibitors 

The cholinergic transmission in the central nervous system is important for the regulation of memory 

and learning process. Numerous studies have highlighted that the cholinergic neurotransmission 

system is profoundly compromised in the brain affected by Alzheimer’s disease (AD), with losses of 

cholinergic neurons and synapses occurring in forebrain, cortex, and hippocampus. The efficacy of 

cholinergic therapies in AD supports the cholinergic hypothesis and validates this neurotransmitter 

system as a therapeutic target. Donepezil inaugurated a new class of acetylcholinesterase (AChE) 

inhibitors with longer and more selective action with manageable adverse effects. It binds both the 

active-site gorge and the peripheral site through benzylpiperidine and indanone moieties. 

In an attempt to obtain analogues of donepezil that could maintain its main spatial and 

physicochemical characteristics while being more compact and less flexible, we applied a strategy to 

synthesize and rapidly evaluate a series of molecules to be eventually developed as AChE inhibitors 

for use in AD. We designed a small library of benzylpiperidinone derivatives, where the indanone 

group was replaced by a bioisosteric indole or pyrrole ring bearing different substituents. The 

biochemical evaluation of the newly synthesized series was performed in collaboration with the 

research group coordinated by Prof. Aldo Roda by using a chemiluminescent (CL) method suitable for 

high-throughput screening (HTS) of AChE inhibitors [1]. The new derivatives showed a rather low 

degree of potency against the enzyme compared to donepezil, but the molecular skeleton allows for 

structural modifications which could be done to explore the potential of the series toward new 

cholinergic agents useful in the treatment of AD [2]. 

To give a further insight into cholinesterase inhibitors, we also prepared new imidazo[2,1-b]thiazole

derivatives [3,4], which were also subjected to CL screening for the evaluation of AChE and 

butyrylcholinesterase (BuChE) inhibitory activity. These studies allowed the identification of some 

analogues displaying a peculiar AChE-selective inhibitory activity in the micromolar range [4]. 

Both AChE and BuChE hydrolyze ACh, albeit with slightly different kinetics, and coexist ubiquitously in 

humans. BuChE is widely distributed in the body of vertebrates and is implicated in neurogenesis and 

regulation of cell proliferation and differentiation. AChE and BuChE inhibitors could reverse the 

cognitive deficits associated with damaging of cholinergic neurons. These enzymes are encoded by 

different genes and clearly differ in substrate specificity and sensitivity to inhibitors, likely due mainly 

to structural differences in the active site gorge. In brain affected by severe AD, the reduction of 

AChE is accompanied by an up to 2-fold increase in BuChE levels. Moreover both enzymes are 

involved in aggregation of beta-amyloid and in formation/maturation of senile plaques, therefore the 

use of reversible cholinesterase inhibitors is considered an attractive therapeutic approach. Unlike 

AChE, the physiologic function of BuChE in normal and diseased humans is not completely clear yet, 

although a role in neurodegenerative disorders has been suggested. Ongoing elucidation of the 

properties and behavior of BuChE in normal and AD brains supports a role for BuChE in AD pathology 

as well as normal cognition. In AD clinical studies the cognitive improvement was correlated with 

rivastigmine (Exelon) treatment. This drug is a well known AChE/BuChE inhibitor and these studies 

support a role for central BuChE in addition to AChE inhibition in modulating cholinergic function. 

Observation made in these studies indicates that a good balance of inhibition of both AChE and 

BuChE may be beneficial in treating the cognitive decline in AD. 

The research we propose concerns the design and synthesis of small libraries of molecules, whose 

AChE/BuChE inhibitory activity will be assayed using the HTS CL method for AChE/BuChE inhibitors 

described above. The availability of a simple, rapid, and sensitive HTS CL method will facilitate the 

study of the biological activity of the designed compounds and the identification of the most 

promising molecules. In addition, the effect of AChE inhibitors could be evaluated “ex vivo” in 

experiment animals by ultrasensitive CL microscope imaging. In fact, the CL system can be also 

employed for the localization and quantification of AChE in brain tissue sections [5]. 

Overall, these studies could furnish new molecular tools useful in elucidating structural and 

functional differences between AChE and BuChE and may contribute to the development of new 

potential drugs for the treatment of AD. 

[1] Guardigli M., Pasini P., Mirasoli A., Leoni A., Andreani A., Roda A.: Chemiluminescent highthroughput 

microassay for evaluation of acetylcholinesterase inhibitors. Anal. Chim. Acta, 535, 139- 

144, 2005. 

[2] Andreani A., Cavalli A., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M., 

Recanatini M., Roda A.: Synthesis and screening for antiacetylcholinesterase activity of (1-benzil-4oxopiperidin-3-ylidene)methylindoles 

and –pyrroles related to donezepil. J. Med. Chem. 44, 4011- 

4014, 2001. 

[3] Andreani A., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M., Roda A.: 

Synthesis and chemiluminescent high throughput screening for inhibition of acetylcholinesterase 

activity by imidazo[2,1-b]thiazole derivatives. Eur. J. Med. Chem., 40, 1331-1334, 2005.

[4] Andreani A., Burnelli S., Granaiola M., Guardigli M., Leoni A., Locatelli A., Morigi R., Rambaldi M., 

Rizzoli M., Varoli L., Roda A.: Chemiluminescent high-throughput microassay applied to imidazo[2,1b]thiazole 

derivatives as potential acetylcholinesterase and butyrylcholinesterase inhibitors. Eur. J. 

Med. Chem., 43, 657–661, 2008. 

[5] Pasini P., Musiani M., Russo C., Valenti P., Aicardi G., Crabtree J.E., Baraldini M., Roda A.: 

Chemiluminescence imaging in bioanalysis, J. Pharm. Biomed. Anal., 18, 555-564, 1998. 


Multidisciplinary projects

Nome Olga Bruno 

Contatti 

obruno@unige.it 

Tel: +39 010 353 8367 

Istituto/Dipartimento Dipartimento di Scienze Farmaceutiche – Università degli Studi di 

Genova 

Viale Benedetto XV,3 16132 Genova 


In the past few years, it has been suggested that dysfunctions of the cAMP/PKA/CREB pathway could be a 

major cause of the cognitive deficits characterizing Alzheimer Disease (AD). Indeed, in vitro experiments 

have shown that Aβ42 is able to reduce the function of this pathway in hippocampal neurons, thus leading 

to impairment of long term potentiation (LTP) and both effects can be reversed by rolipram, a well known 

PDE4 inhibitor. More interestingly, rolipram has been shown to restore LTP and to ameliorate cognitive 

deficits in a double transgenic (APP/PS1) murine model of AD. Recently, sub-chronic rolipram treatment 

was reported to improve long-term memory performances also in normal rodents. 

Despite PDE4 inhibitors have been indicated as promising therapeutical agents for AD, rolipram cannot be 

clinically used due to its adverse effects, particularly sedation and nausea. Among the different PDE4s, 

PDE4D is highly expressed in hippocampal regions and some isoforms seem to be augmented in the 

hippocampus of AD patients. 

Recently we have synthesized a series of innovative selective PDE4D2, PDE4D3 inhibitors, 1 which increase 

cAMP levels in the hippocampus and improve cognitive functions in preliminary in vivo pre-clinical studies. 

Surprisingly, in vitro experiments showed that our compounds, as well as rolipram, augment Aβ levels in 

neuronal cultured cells, a result in agreement with a recent study showing that dimebolin, a drug known to 

ameliorate cognitive deficits in aged rodents and in people suffering from mild AD, causes an acute 

increase of Aβ brain levels in experimental animals. 

Our project is aimed at studying new PDE4D selective inhibitors as novel therapeutic agents in AD. These 

compounds could be also useful pharmacological tools to increase the knowledge on the role of different 

PDE4 isoforms in CNS under physiologic as well as pathologic conditions, and to verify the relationship 

between PDE4D inhibition, A� levels and memory improvement. 

Our research unit skilful in medicinal chemistry will work to modify the chemical structure of previously 

synthesized compounds in order to increase its selectivity, potency and in vivo activity. Our first goal is to 

obtain a beneficial effect on memory performances without emesis. 

The molecular modifications will be performed basing on a preliminary computational study aimed to 

show what structural features could discriminate between different PDE4D splicing variants. Thus docking 

studies and 3D-QSAR analysis will be used to filter the large number of new molecular entities deriving 

from various chemical modifications on our previously synthesized compounds. 

1) Bruno O, Romussi A, Spallarossa A, Brullo C, Schenone S, Bondavalli F, Vanthuyne N, Roussel C. New 

selective phosphodiesterase 4D inhibitors differently acting on long, short, and supershort isoforms. J Med 

Chem. 2009 52(21), 6546-57. 


Drug Design and synthesis of therapeutic agents for neurodegenerative 

disorders, particularly for Alzheimes Disease. 


Titolo progetto Heterocycle compounds as neuroprotective agents 


(MIUR, Italian Minister of University Research) 2005032713_005 

Ronsisvalle Giuseppe (Principal Investigator)

Bruno Olga Role: Co-investigator 


Abstract del progetto This study was aimed at synthesizing new molecules acting by different 

pathways in neurodegenerative pathologies. 

Titolo progetto Heterocycle compounds as neuroprotective agents 


(MIUR, Italian Minister of University Research) 2007E8CRF3_003 

Ronsisvalle Giuseppe (Principal Investigator) 

Bruno Olga Role: Genoa Unit PI 


Abstract del progetto This grant is a renewal of previously financial support for a large study 

aimed to synthesize new molecules acting by different pathways in 

neurodegenerative pathologies.

Nome Fabrizio Tagliavini 

Contatti 

Tel +39 02 2394 2260; Fax +39 02 2394 2101 

Email: ftagliavini@istituto-besta.it 

Sito web: http://www.istituto-besta.it 

Istituto/Dipartimento UO Neuropatologia – Neurologia 5 


Via Celoria ,11 - 20133 Milano- ITALY 


The research group: The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the 

diagnosis and treatment of patients with degenerative dementias (over 2000 out-patients and 150 inpatients 

per year) and (ii) a Laboratory Unit (certified ISO 9001:2000 - Registration Number 26080, 

dedicated to the analysis of genes and biomarkers associated with degenerative dementias, postmortem 

characterization of the disease process and disease-specific protein, and experimental 

studies on disease pathogenesis and the development of therapeutic strategies. The course of action 

of this activity can be expressed as a “Bed to Bench to Bed” cycle, with the ultimate goal to identify 

disease-modifying drugs in preclinical settings and apply them to patients. In this regard, the Clinical 

Unit is currently coordinating a multicentre phase II clinical trial supported by AIFA, to test the 

effectiveness of an anti-amyloidogenic molecule identified in experimental models. This 

comprehensive approach has also the great advantage to collect biological samples (plasma, DNA, CSF 

and brain tissue) from fully characterized patients. In this regard the Laboratory Unit is part of the 

Network of Excellence “BrainNet” Europe devoted to biobanking, harmonization of assessment tools 

and standardization of diagnostic criteria of neurodegenerative diseases. 


comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1 

technician who are committed to the research activities on degenerative dementias with different 

expertise and roles. On the overall, the team has large experience in a variety of techniques spanning 

from neuropathology (including immunohistochemistry, morphometry, electron microscopy and 

atomic force microscopy), to molecular genetics, biochemistry (purification and characterization of 

disease-specific proteins), cellular and molecular biology, and animal models. The Division of 

Neuropathology is provided with fully equipped laboratories for histology, immunohistochemistry, 

electron microscopy, atomic force microscopy, biochemistry, and molecular and cellular biology, and 

a 60 sqm BL3 facility. Moreover an animal facility composed by a 140 sqm environmental controlled 

area, provided with a HVAC system and heap-filtered ventilated racks, with separate rooms for 

maintaining and breeding mouse lines, and carrying out surgical procedures, collection of samples 

and post-mortem examination is available. 


• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic 

strategy for sporadic AD 

We have recently identified an APP mutation (A673V) that causes early-onset AD only in the 

homozygous state while the heterozygous carriers are not affected. This mutation strongly boosts the 

production and amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and 

wild-type peptides inhibits amyloidogenesis and Aβ-mediated neurotoxicity (Di Fede et al., Science 

2009, 323:1473-7). These findings are consistent with the observation that the A673V heterozygous 

carriers do not develop disease and offer grounds for a novel therapeutic strategy based on modified 

Aβ peptides. A research priority of our lab is to unravel the molecular mechanisms of the opposite 

effects of the A673V APP mutation in homo- or heterozygous state on amyloidogenesis and to develop 

a lead compound for AD therapy based on A673V-modified Aβ peptides or peptido-mimetic 

molecules. To accomplish these objectives we have generated a panel of transfected cells and 

transgenic C. elegans expressing human APP or Aβ with the A673V mutation, respectively, and

transgenic mouse lines expressing A673V-mutated APP in the homozygous or heterozygous state. 

Furthermore, we have identified a prototypic lead compound corresponding to a six-mer Aβ peptide 

with the A673V substitution and are currently working on brain delivery systems. 


identificata 

CLINICA/PRECLINICA 

- Genetic susceptibility to Alzheimer’s disease (AD) 

- Early diagnosis 

- Biomarkers 

- Developing competitive animal models to study AD 

- Studying early onset forms of AD to follow-up disease progression 

- Cinical trials with or without involvement of pharmaceutical companies 


- Biobanking (blood sample, CSF, brain repository…) 

- Standardization of diagnostic criteria 


- Translational research

Nome Simone OTTONELLO 

Contatti 

+39 0521 905646 (office) 

+39 0521 905148 (lab) 

s.ottonello@unipr.it 

Istituto/Dipartimento Department of Biochemistry and Molecular Biology 

Viale G.P. Usberti 23/A 


43100 Parma (Italy) 


Area di interesse identificata -“Biomarkers” / “Early diagnosis” 

Implementation of high-resolution techniques for A��oligomer detection 

and quantification in brain samples from Tg-mouse models of Alzheimer 

disease and in body fluids, including human CSF. 





-“Basic research” 

Yeast mutant strains (selected from a genome-wide mutant collection) 

conditionally expressing Alzheimer-related pathogenic polypeptides as 

tools to unravel new fundamental aspects of AD pathogenesis (e.g., 

mitochondrial function impairment) and potential treatments thereof. 

-“New treatment strategies” 

Second generation, recombinant A� peptide immunogens as prototype 

vaccines for AD treatment/prevention (active immunization approach) 

and as primary antigens for the production of conformation-sensitive 

therapeutic monoclonal antibodies (passive immunization approach). 

“Novel anti-A� immunization approaches for the treatment of Alzheimer 

disease” 

Pharmaceutical Company 

Titolo progetto “Further studies on the Trx-A� antigen and its potential as an anti-AD 

vaccine” 



Durata progetto ��year 

Titolo progetto “Target selectivity of NSAID �-secretase modulators evaluated by oligomacroarray 

analysis” 



Durata progetto ��year 

Titolo progetto "Large-scale production of recombinant TrxA� for a preclinical study in

Tg-AD mice” 

Ente finanziatore Pharmaceutical Company 

Durata progetto ��months 

Titolo progetto "Immunoelectrophoretic analysis of A� oligomers in brain samples from 

Tg-AD mice treated with a NSAID �-secretase modulator” 

Ente finanziatore Pharmaceutical Company 

Durata progetto 6 months (ongoing)

Nome MAURIZIO SCARPA 

Contatti Maurizio.scarpa@unipd.it 

Istituto/Dipartimento DEPARTMENT OF PEDIATRICS UNIVERSITY OF PADOVA ITALY 



identificata 






PEDIATRIC NEURODEGENERATIVE DISEASES 

BRAINS4BRAIN – Treating Pediatric Neurodegenerative Diseases: 

From Laboratory Bench to Bedside 

EUROPEAN SCIENCE FOUNDATION 

Exploratory Workshop March 3-5 2010 Frankfurt 

Neurodegenerative diseases are most prevalent in the elderly, but can in rare cases also 

affect individuals early in life. In children, neurodegeneration leads to severe mental 

retardation and premature death with devastating consequences on their immediate 

environment and relatively high costs for society. Within the EU pediatric 

neurodegenerative disorders are considered “Rare”. Because of the low-prevalence of 

these disorders, there is often a striking lack of information, research, treatment and 

expert availability. There is also often a significant delay before a definitive diagnosis is 

achieved. BRAINS4BRAIN calls for a collaborative research effort to focus the available 

multidisciplinary European brainpower with relevance to tackling pediatric 

neurodegenerative diseases. BRAINS4BRAIN (www.brains4brain.eu) is a European task 

force aimed at developing and implementing new therapies for pediatric 

neurodegenerative diseases for which, at the moment, no effective therapy is available. 

The task force wishes to create a coordinated European effort towards i) the 

comprehension of pathophysiology processes of these pediatric neurological disorders, ii) 

the implementation of knowledge on the BBB, iii) the development of new molecular 

and/or biochemical strategies to overcome the BBB for brain-targeted drug delivery, and 

iv) the development and implementation of diagnostics and clinical therapies to timely 

detect and treat these devastating CNS disorders. Importantly, it is foreseen that such 

knowledge and insights will also be valuable for understanding and treating other 

important neurological diseases such as Alzheimer’s, Parkinson’s Disease, and epilepsy. 

LSDs are metabolic disorders, caused by the lack of certain (lysosomal) enzymes or 

lysosome components, thus preventing the complete degradation of macromolecules and 

the recycling of their components. 

The accumulation of intermediate degradation products affects the appropriate 

functioning of lysosomes and other cellular organelles. 

Accumulation starts immediately after birth and progressively worsens, often affecting 

several organs, including the Central Nervous System (CNS). CNS pathology causes mental 

retardation and progressive neurodegeneration that ultimately ends in early death of 

these young patients. LSDs are the only group of pediatric neurodegenerative diseases 

for which therapy that can reverse the natural history of the disease in peripheral organs 

is available (Enzyme Replacement Therapy, ERT). 

Unfortunately, ERT is currently unable to effectively reach the CNS to stop the lethal 

progression of the neurodegeneration. Nonetheless, ERT in combination with i) the 

advanced knowledge of the (genetic- and biochemical) causes for the development of 

neurodegeneration in LSDs and ii) the availability within Europe of well established in 

vitro and animal models, now provides the unique opportunity to decipher the cascade of 

events leading to loss of brain plasticity and mental retardation, as well as its possible 

reversal. For instance, suppressing the primary cause of neurodegeneration (e.g. by ERT) 

in a young brain –that at this stage of development retains considerable plasticity–

maximizes the potential for neurological repair. Important to note is that common 

secondary events in both pediatric and adult neurodegenerative diseases lead to 

neurodegeneration. Studies on LSDs therefore have the ability to provide unique insights 

into the pathophysiology and restorative capacities of neurodegenerative diseases in 

general. Therapies based on ERT can modify the natural history of some LSDs in the 

peripheral organs (liver, spleen, joint mobility etc.). However, the BBB prevents the 

therapeutic enzymes used from reaching the CNS and modifying the course of 

neurodegeneration 1. This limitation can be circumvented by non-pharmaceutical therapy 

in which sources of enzyme (e.g. a viral vector) are created by surgical injection directly 

into the CNS. It might eventually also be achieved to some extend by pharmaceutical 

therapies based on BBB traversing small molecules aimed at reducing the accumulation of 

molecules requiring degradation (e.g. Substrate Reduction Therapy (SRT):inhibiting early 

enzymes in the biosynthetic pathway, or Chemical Chaperones: assisting the affected 

enzyme to attain its correct, active conformation2).3,4 However, ERT is likely to be the 

most efficient strategy with the least side effects. As neurodegeneration in LSDs affects 

the entire brain, the technological challenge is to safely deliver ERT efficiently to all 

affected areas. This might be achieved only by developing strategies enabling the 

therapeutic enzyme to cross the BBB. Tools to enable the entry of curative molecules 

across the BBB into the CNS will represent a major breakthrough for the long-term 

therapy of many (if not all) CNS diseases. Several cutting edge initiatives using different 

approaches to specifically deliver molecules across the BBB (e.g. exploiting certain 

transporters or manipulating BBB permeability) exist within Europe.

Nome Paola Tirassa 

Contatti 

p.tirassa@inmm.cnr.it; INMM- CNR Istituto di Neurobiologia e Medicina 

Molecolare Via del Fosso di Fiorano, 64 00143 ROMA ITALY Phone: + 39 

06 501 703 236 Fax: + 39 06 501 703 313 

Istituto/Dipartimento Institute of Neurobiology and Molecular Medicine CNR 


2) Basic research : New treatment strategies based on Neurotrophins 

f) Ocular NGF administration as a novel non invasive approach to protect brain-NGF target neurons that 

degenerate in Alzheimer’s disease (L. Aloe, P. Tirassa). Nerve growth factor and (NGF) is a soluble protein 

that plays a protective role on brain neurons that degenerate in age-related brain disorders, including 

Alzheimer’s disease (AD). We have shown that conjunctively applied NGF can protect injured brain 

neurons and damaged retinal ganglion neurons suggesting that topical eye NGF application might be a 

novel alternative for delivering NGF into the brain and for protecting brain neurons and reducing memory 

deficits occurring during early events of AD and glaucoma. This hypothesis is currently actively under 

investigation in collaboration with other research groups. 

Lambiase A, Pagani L, Di Fausto V, Sposato V, Coassin M, Bonini S, Aloe L. Nerve growth factor eye drop 

administrated on the ocular surface of rodents affects the nucleus basalis and septum: biochemical and 

structural evidence. Brain Res. 2007;1127:45-51. 

Di Fausto V, Fiore M, Tirassa P, Lambiase A, Aloe L. Eye drop NGF administration promotes the recovery of 

chemically injured cholinergic neurons of adult mouse forebrain. Eye drop NGF administration promotes 

the recovery of chemically injured cholinergic neurons of adult mouse forebrain. Eur J Neurosci. 

2007;26:2473-80. Lambiase A, Aloe L, Centofanti M, Parisi V, Mantelli F, Colafrancesco V, 

Manni GL, Bucci MG, Bonini S, Levi-Montalcini R. Experimental and clinical evidence of neuroprotection by 

nerve growth factor eye drops: Implications for glaucoma. Proc Natl Acad Sci U S A, 2009; 109: 13469- 

13474,. 

g) Electro-acupuncture and polyphenols as novel approaches to reduce neurodegeneration ( L Manni, M 

Fiore). Preliminary data indicate that diabetes induced in adult rats by injection with streptozotocin causes 

a decrease of brain NGF and a concomitant increase of tau phosphorylation, that were all counteracted by 

low-frequency electro-acupuncture (EA). Using molecular and behavioural approaches the research 

project will be aimed at: i) Investigate the possible link between diabetes, brain NGF presence/activity and 

the development of AD-associated pathological features; ii) Study the effect of EA on the above mentioned 

diabetes-associated brain dysfunction. Alcoholism is related to neurodegeneration and diabetes. The use 

of polyphenols extracted by olive oil, olive leaves and olive seeds or by grapes (Fiore et al 2009) to prevent 

or limit mouse brain neurodegeneration due to aging or ethanol consumption will be evaluated. 

Manni L, Aloe L, Fiore M. Changes in cognition induced by social isolation in the mouse are restored by 

electro-acupuncture. Physiol Behav. 2009;98(5):537-42 

Fiore M, Laviola G, Aloe L, di Fausto V, Mancinelli R, Ceccanti M. Early exposure to ethanol but not red wine 

at the same alcohol concentration induces behavioral and brain neurotrophin alterations in young and 

adult mice. Neurotoxicology. 2009;30(1):59-71. 

h) Human neural stem cells lentivirally transduced with human BDNF (C. Cenciarelli, P. Casalbore). A 

strong body of evidences show that BDNF result critically involved in Alzheimer’s and Huntington’s 

diseases. Recently, it has been shown that BDNF knockdown within NSC abolishes the cognitive benefits of

NSC delivery. The aim is to use propagating human neural stem cells (hNSC) lentivirally transduced with 

human BDNF as cellular therapy for replacing degenerating neurons in disease, trauma and toxic insults. 

Cenciarelli C, Budoni M, Mercanti D, Fernandez E, Pallini R, Aloe L, Cimino V, Maira G, Casalbore P. In vitro 

analysis of mouse neural stem cells genetically modified to stably express human NGF by a novel multigenic 

viral expression system. Neurol Res. 2006, 28:505-12. 

Casalbore P, Barone I, Felsani A, D’Agnano I, Michetti F, Maira G and Cenciarelli C. Neural stem cells 

modified to express BDNF antagonize trimethyltin-induced neurotoxicity through PI3K/Akt and MAP kinase 

pathways . Accepted on J. Cell. Physiol. 2010.

Nome Micaela Morelli 

Contatti 

Istituto/Dipartimento Department of Toxicology, University of Cagliari 








New symptomatic and neuroprotective therapies for Parkinson’s 

disease 

1) Development and dissemination of innovative methodologies 

for planning, synthesis and determination of biological activity 

of new ligands for membrane receptors (FIRB:RBNE03YA3L_004) 

2) Evaluation of behavioural and biochemical changes in 

'Engrailed' mutant mice (PRIN 2006) 

3) : Evaluation of neuroprotective effects of ST1535 in animal 

model of Parkinson ‘s disease (SIGMA-TAU industry) 

1-2) MURST 

3 ) SIGMA-TAU Industry 

1) 3 years 

2) 2 years 

3) 1 year 

1) To determine the in vivo and ex vivo biological activity of 

adenosine A2A antagonists compounds of new synthesis. Three 

compounds symptomatically active on models of Parkinson’s 

disease have been selected on the basis of the results obtained. 

Moreover their potential neuroprotective effects has been 

recently investigated in models of Parkinson’s disease. 

2) The engrailed strain of mice has shown behavioural deficits and 

biochemical changes superimposible of Parkinson’s disease 

models. This strain of mice is now used as a model of this disease. 

3) Evaluation of neuroprotective and anti-inflammatory effects of 

new adenosine A2A receptor antagonist ST1535 in animal model 

of Parkinson ‘s disease. ST1535 has shown positive symptomatic 

improvement in motor functions and also neuroprotective and 

anti-inflammatory effects in the MPTP mouse model of 

Parkinson’s disease.

Nome Maria P. Abbracchio 

Contatti 

Tel. 

E mail 



Dept Pharmacol Sci, Univ Milan, Italy 

+39-02-50318310/355 

mariapia.abbracchio@unimi.it 

Area di interesse identificata Innovative neuro-reparative strategies via the implementation of 

endogenous neurogenesis and gliogenesis: focus on the new P2Y-like 

GPR17 receptor 








Abstract delle ricerche in 

corso 

PURINOCEPTORS AND NEUROPROTECTION: FOCUS ON THE NEW 

PURINERGIC RECEPTOR GPR17 

COFIN-MIUR 2008; 

2 anni 

IMPLEMENTATION AND GLIOGENESIS VIA THE PURINERGIC SYSTEM: A 

NEW STRATEGY TO REPAIR ACUTE NEURODEGENERATIVE DISEASE 

Ministero della Salute 2007 

2 anni 

Deciphering the molecular mechanisms controlling the behaviour of 

endogenous neural stem cells is of paramount importance to understand 

the processes at the basis of brain recovery and to foster the brain’s 

ability to repair itself in acute and chronic neurodegenerative diseases. 

We have recently identified a new P2Y-like G-protein-coupled 

purinoceptor GPR17, activated by both uracil nucleotides and cysteinylleukotrienes 

(cysLTs) (Ciana P et al., The orphan receptor GPR17 identified 

as a new dual uracil nucleotides/cysteinyl-leukotrienes receptor. EMBO J 

25:4615-27, 2006). We have established a national consortium (the 

GPR17 research team) that specifically studies the role of this receptor in 

cerebral diseases. Initial data in rodent models of focal brain ischemia 

have shown that GPR17 participates to both the propagation of brain 

damage and to the subsequent post-injury remodelling and repair (Lecca 

D et al., The recently identified P2Y-like receptor GPR17 is a sensor of 

brain damage and a new target for brain repair. PLoS ONE 3:e3579, 1-15, 

2008). Specifically, GPR17 was found to activate the quiescent 

oligodendrocyte precursor cells that are still present in the adult brain, 

inducing them to resume myelination, thus re-establing neuron-toneuron 

communication. GPR17 was also found to be expressed by the 

ependymal cells lining spinal cord’s central canal (Ceruti S et al., The P2Ylike 

receptor GPR17 as a sensor of damage and a new potential target in 

spinal cord injury. Brain 132:2206-18, 2009) that are believed to 

represent the true adult stem cells in this part of the central nervous

system. Upon spinal cord mechanical injury, these cells started 

proliferating and re-expressing markers of multipotent stem-like cells 

(ibidem). Interestingly, in the adult brain, GPR17 specifically decorates a 

precursor cell (NG2-positive neural precursors) that normally 

differentiates to oligodendrocytes, but that, under appropriate 

conditions, can be addressed to generate functional new neurons and 

new astrocytes. These data make GPR17 an interesting new target not 

only for stroke but also for chronic human neurodegenerative diseases 

characterized by demyelination or neuronal pathology, such as multiple 

sclerosis (MS) and Alzheimer’s disease (AD). Preliminary data in animal 

models indeed support this hypothesis: activated GPR17-expressing adult 

precursor cells were found to accumulate, respectively, around 

demyelinated wounds in an Experimental Autoimmune Encephalomyelitis 

(EAE) MS animal model, and around brain plaques in a transgenic murine 

model of AD (APPPS1 mice). 

Our current efforts aim at: 

1. We want to develop an in silico tridimensional pharmacophore model 

of GPR17 to screen large virtual chemical libraries and to design new 

specific ligands. The newly synthesized agonists/antagonists (including 

new “dual” ligands acting at both the nucleotide and the cysLT binding 

sites) will be tested in our established in vitro assays and then addressed 

to their in vivo validation 

2. “Old” and new GPR17 ligands will be used to unveil the exact role of 

the receptor during oligodendrocyte differentiation and myelination, in 

both in vitro and in vivo models, and to identify the factors regulating its 

transcription in precursor cells. 

3. To set the basis for a possible wider exploitment of GPR17 in adult 

neurogenesis/gliogenesis, by using both isolated precursor cells and 

neurospheres from control and diseased brains, we shall also develop 

pharmacological protocols to address GPR17-positive precursor cells to 

neurons or astrocytes. 

4. The role of GPR17 in the EAE MS model will be studied in detail. Initial 

information on the changes of GPR17 expression in these models will be 

obtained via an in silico analysis of microarray data; then, the regional 

localization, cell types, and kinetics of GPR17 expression during disease 

evolution will be assessed. Finally, we shall determine the effects of a 

“preventive” or “therapeutic” in vivo administration of GPR17 ligands on 

animals’ weight, neurological score and disease course. 

Globally, results will contribute to finding new GPR17-based regenerative 

medicine approaches for human neurodegenerative diseases, allowing for 

substantial improvement of “endogenous neural stem cell”-based in vivo 

therapy.

Nome Mariagrazia Grilli/Pier Luigi Canonico 

Contatti 

Tel. 

E mail 

Mariagrazia Grilli 

0321375828 3280587577 

grilli@pharm.unipmn.it 

Istituto/Dipartimento DiSCAFF, University of Piemonte Orientale, Novara Italy 



identificata 






Novel drugs (D). 

“Pharmacological modulation of adult neurogenesis” 

Pharmacological modulation of endogenous neural stem cells for 

amelioration of cognitive impairment in neuropsychiatric 

disorders 

Cariplo, Regione Piemonte, PRIN, Fondazione Comunità del Novarese 

3 anni 

As of today, the pharmacological treatment of cognitive deficits associated with 

several neuropsychiatric disorders is still very limited. The essential requirement 

for an ideal anti-dementia drug would be the ability to re-organize the neuronal 

network in the injured brain so as to prevent/slow down the neurodegenerative 

process and possibly to partially recover brain function. Structural and functional 

plasticity, often referred to as “neuroplasticity”, a fundamental property of the 

brain, comprises chemical, electrical, molecular and cellular responses by which 

connections within a brain region and/or between brain regions are reorganized. It 

is maximal during brain development but it persists to a relevant extent in 

adulthood, conferring to neuronal networks a higher degree of circuit adaptation 

to the novel experiences we are exposed to for our entire life. Neuroplasticity in 

adulthood is in fact the neurobiological substrate of learning and memory. A 

better understanding of molecular events underlying neuroplasticity may lead to 

innovative therapeutic approaches in a variety of neuropsychiatric disorders 

characterized by learning and memory deficits. 

Different forms of neuroplasticity are known to exist in the adult brain, 

including neurogenesis. A large body of experimental work has recently 

established that newborn neurons continue to be added to discrete regions of the 

adult nervous system including the hippocampus, an area implicated in learning 

and memory which form the basis for complex behaviour patterns in vertebrates. 

Several studies support the hypothesis that experience- or activity-dependent 

integration of new neurons into hippocampal pre-existing neural networks 

represents a functional mechanism of brain plasticity subserving specific types of 

learning and memory functions. Several human disorders affecting memory and 

cognition, including neurodegenerative and psychiatric disorders, are 

characterized by abnormalities in synaptic structure/number as well as in 

neurogenesis, including Alzheimer’s disease (AD). The present project aims to 

investigate whether the pharmacological manipulation of neuroplasticity in 

pathological conditions may be effective in modulating different memory stages 

and systems. This would be a starting point for developing and testing novel

pharmacotherapies for cognitive disorders. In particular, the contribution of NFkappaB 

proteins to this process and in cognitive disorders is under investigation. 

Indeed a vast body of information points at the role of the NF-kappaB family in 

synaptic plasticity, learning and memory and at neuropychiatric disorders arising 

when it is deranged (Grilli & Meneghini, in press, 2010). Recently, we showed 

that NF-kappaB p50KO mice displayed a very selective defect in hippocampal - 

dependent short-term memory which correlated with the disruption of 

hippocampal adult neurogenesis (Denis-Donini et al., 2008). More specifically, in 

the absence of p50 protein, defective neurogenesis resulting in memory defects 

appeared to possibly occur at the transition between distinct maturation steps in 

newly generated neurons. Intriguingly, this is the stage characterized by high 

synaptic plasticity when in the adult newborn neurons with extended axons and 

dendrites have to be selected so as to become integrated into the preexisting 

functional network. Morover, we were also able to show that several drugs 

promoting in vitro and in vivo neurogenesis actually involve the NF-kappa 

signalling pathway. Altogether, these data identify NF-κB signaling components 

as critical mediators of adult neurogenesis and suggest that that they may 

represent previously undescribed therapeutical targets for neuropsychiatric 

disorders, including AD. The overall goal of this research proposal is evaluate 

whether a disregulated NF-kappaB signalling may affect mechanisms regulating 

neuronal maturation and differentiation of adult generated neurons and, as a 

consequence, contribute to cognitive impairment associated with relevant CNS 

disorders. To this aim, we will use a relevant animal model of AD such as the 

TgCRND8 mouse line as well as the p50KO mice, both characterized by cognitive 

impairment. Experimental strategies will be applied to allow to prove the concept 

that this signaling pathway can be pharmacologically modulated to promote adult 

neurogenesis and, in turn, ameliorate cognitive impairment. Our project has the 

potential to increase our understanding of the molecular participants in the 

modulation of adult neurogenesis in physiology and pathology and it also holds 

the potential for identifying novel strategies aimed at ameliorating cognitive 

impairment associated with CNS disorders of high medical need but with no or 

unsatisfactory therapy.

Nome Fabrizio Chiti 

Contatti 

Tel. 

E mail 

Dip. di Scienze Biochimiche, Univ. di Firenze, V.le Morgagni 50, Firenze. 

Tel: +39-055-4598319 

e-mail: fabrizio.chiti@unifi.it 

Istituto/Dipartimento Dip. di Scienze Biochimiche, Università di Firenze, Viale Morgagni 50, 

Firenze 


Area di interesse identificata We intend to focus on the relationship structure-function of oligomers of 

the β amiloid peptide and its analogues. We will use a number of 

methodologies to gain structural information on the oligomers at the 

molecular level, including site-directed mutagenesis, site-directed 

labelling with molecular probes to be followed in fluorescence, NMR and 

EPR. We will also use cultured cells and animal models, evaluating the 

effect of structural changes of the oligomers on their toxicity, of inhibitors 

of toxicity such as chaperones and small molecules, of changes of the 

membrane lipid content, of the presence of glycosaminoglycans and 

other components of the extracellular matrix, and of protective agents 

such as extrogens, IGF-1 and others that mediate neuroprotection via the 

activation of the seladin-1 gene. We will use common cellular cultures but 

also cultures of primary neurons of rat and human ippocampi to evaluate, 

for example, interactions between oligomers and synapses or oxidative 

stress of the membrane. We will also use animal models such as 

transgenic mice. The following research groups will participate: 

Prof. Fabrizio Chiti Dipartimento di Scienze Biochimiche 

Prof. Massimo Stefani Dipartimento di Scienze Biochimiche 

D.ssa Cristina Cecchi Dipartimento di Scienze Biochimiche 

Prof. Fiorella Casamenti Dip. di Farmacologia Precl. e Clinica 

Prof. Alessandro Peri Dipartimento di Fisiopatologia Clinica 


Titolo progetto The role of cholesterol in the pathogenesis of Alzheimer's disease 


Fondazione Cassa di Risparmio di Pistoia e Pescia and Centro 

Internazionale delle Malattie neurodegenerative (CIMN) 

Durata progetto 12 months (January 2010 - December 2010) 

Abstract del progetto We will investigate the relationship between membrane cholesterol 

content and cytotoxicity induced by Aβ oligomers on human 

neuroblastoma cells and primary fibroblasts from familial AD patients 

bearing genetic mutations. 


Cytotoxicity of amyloid aggregates in neuronal cells and in Alzheimer 

fibroblasts: effect of putative anticytotoxic molecules. 

Ente finanziatore MIUR and Università di Firenze (cofinanziamento PRIN 2008) 

Durata progetto 24 months (March 2010 – March 2012) 

Abstract del progetto We will study the ability of antioxidant compounds, such as glutathione 

thioesters (S-acyl-GSH) derivatives, to prevent the degenerative effects 

triggered by Aβ accumulation on primary cortical neurons, human 

neuroblastoma cells and primary skin fibroblasts from familial AD

patients. 


Effect of the extracellular matrix on the formation, stability and 

cytotoxicity of amyloid fibrils and their oligomeric precursors 



Abstract del progetto We will study the effect of components of the extracellular matrix where 

amyloid accumulate (chaperones, glycosaminoglycans, collagen fibres, 

etc.) on the formation, stability and toxicity of amyloid fibrils and their 

precursors 


Study of the structure-toxicity relationship of protein oligomers 

Ente finanziatore EMBO Young Investigator Programme 

Durata progetto From 2005 to end of funding (still available to date) 

Abstract del progetto We will investigate the relationship between the structure of protein 

oligomers formed by a model proteins at the molecular level and their 

biological effect on cultured cells, primary neurons and animal models 


Structural and function aspects of lipid bilayers in molecular medicine: 

involvement of lipid rafts in human pathology 

Ente finanziatore Cassa di Risparmio Firenze 

Durata progetto 36 months (2008-2010, but estende to one more year) 


The project will investigate the role of natural or synthetic llipid surfaces 

with different lipid composition and physicochemical features as 

modulators of peptide/protein misfolding and aggregation as well as of 

amyloid toxicity. 


Integrative approach to the interplay between Aß, oxidative stress, the 

proteasome complex and the autophagic pathway. Effect of 

neuroprotective molecules 





To characterize in vivo in the TgCRND8 mouse model of ß-amyloid 

deposition the molecular bases correlating oxidative damage and 

autophagy and ubiquitin-proteasome dysfunctions to the gradual 

deposition of Aß and to evaluate the effects of neuroprotective drugs. 

Design of small molecule therapeutics for treatment of Alzheimer's 

disease based on the discovery of innovative drug targets. 

Ente finanziatore European commission (6FP, ADIT) (project LSHB-CT-2005-511977) 

Durata progetto 5 years (June 2005-November 2010) 



To analyze in vivo the expression and phosphorylation state of new 

protein targets in selected brain regions of mouse models of Aβ 

overproduction using standard immunohistochemical methodologies. To 

analyze phenotype of target over-expression in transgenic mice, generate 

inside the project, with respect to neurodegenerative and cognitive 

parameters 

Oleuropein aglycon and hydroxythyrosol as anti-aggregating molecules 

potentially exploitable in Alzheimer’s disease prevention and therapy

Ente finanziatore Regione Toscana 

Durata progetto 4 months (November 2010 – October 2012) 


The research will study both in vitro and in vivo the anti-aggregating 

effect on Abeta42 of oleuropein aglycone and its derivative 

hydroxytyrosol, to assess the possible use of these molecules for 

prevention and treatment of Alzheimer’s disease.


E- Non-Pharmacological Therapies


Contact person: Pio E. Ricci Bitti, M.D. (Dept. of Psychology, University of Bologna) 

pioenrico.riccibitti@unibo.it 

Date: January 29, 2010 

I) Strategic Issues ( Justification of the importance of the issue ) 

2.2.1 Brain and brain-related diseases 

An increasing number of Parkinson disease (PD) patients are undergoing deep brain stimulation DBS). 

It is now important to evaluate not only the benefits concerning the main symptoms related to movement, 

but also the psychological (cognitive, affective and behavioural) impact of DBS. 


In relation to our experience with PD patients, the most important area to be evaluated by a psychological 

point of view is the possible influence of DBS on cognitive processes (such as memory, spatial orientation, 

time perception and perspective…). 

III) Impact of the research area 

The results of this kind of studies could be used in order to propose specific 

psychoeducational interventions to improve the cognitive efficacy of PD patients that underwent DBS

Nome Maurizio Taglialatela, MD PhD – Professor of Pharmacology 

Claudio Russo, PhD – Associate Professor of Pharmacoloy 

Alfonso Di Costanzo MD – Associate Professor of Neurology 

Giovanni Scapagnini MD PhD – Associate Professor of Clinical 

Biochemistry 

Contatti 

Email: m.taglialatela@unimol.it 

Tel. +39-0874-404894/4851/4891 

Istituto/Dipartimento Dept. of Health Science, University of Molise 

Via De Sanctis, 8610 Campobasso - ITALY 



identificata 


Titolo 

progetto 

Ente 


Durata 

progetto 

Abstract del 

progetto 

Physical exercise and cognitive decline. Epidemiological studies have shown that 

physical exercise can delay the occurrence and the progression of dementia, but also to 

improve physical, cognitive and psychological performances, with a positive impact on 

the quality of life. The mechanisms involved and the identification of potential 

biomarkers predictive of a positive response to physical activity are areas of intense 

investigation. In our group, we are currently investigating the potential preventive role of 

physical exercise programs on cognitive decline in subjects at risk to develop dementia. 

Our focus will be on subjects presenting with a subjective memory disturbance or 

affected by mild cognitive impairment (MCI). 

• Tedeschi G., et al. Brain atrophy and lesion load in a large population of patients with 

multiple sclerosis. Neurology. 2005 Jul 26;65(2):280-5. 

• Tedeschi G., et al. Correlation between fatigue and brain atrophy and lesion load in multiple 

sclerosis patients independent of disability. J Neurol Sci. 2007 Dec 15;263(1-2):15-9. 

Regulation of K + channels by APP-interacting proteins: functional characterization and 

implications for AD-related neurodegeneration 

Regione Campania (2000-2003) 

3 years 

Recent experimental evidence suggest that K + channels play a major pathogenetic role in 

Alzheimer disease. In the CNS, phosphorylation of voltage-gated K+ channels by tyrosine 

kinases (PTK) is a fundamental process regulating their functional activity; its inhibition exerts a 

neuroprotective action in several experimental models of neuronal damage. On the other hand, 

the expression of several subclasses of K + channels can reduce cellular tyrosine kinase activity, 

thus suggesting the existence of a reciprocal interaction between PTKs/PTPases and K + 

channels. Given that the cytosolic domain of β-APP influences, through its adaptor proteins 

Dab (disabled), X11 and Fe65, the tyrosine kinase activity of Abl, in the present proposal we 

will: 

E. Evaluate the potential modulation of several classes of K + channels (Kv1.2, Kv1.3, Kv1.5, 

Kv2.1, ERG, KCNQ2+3) by adaptor proteins able to bind to β-APP (X11, Fe65, Dab). 

F. Verify the participation of tyrosine kinase activity (and in particular of Abl), in this 

modulation. 

G. Investigate the potential involvement of these channels in the cellular damage induced by 

the hyperexpression of β-APP adaptor proteins. 

H. Study the participation of K + channels and their regulation by oxidative stress in the 

neuronal death triggered by various cellular models of AD-related neurodegeneration. 

Titolo European Community contract N° LSHM-CT-2003-503330/Apopis : Abnormal proteins in the

progetto 

Ente 


Durata 

progetto 

Abstract del 

progetto 

pathogenesis of neurodegenerative disorders 

Study on tyrosine phosphorylation of APP and its interaction with intracellular adaptors: role in 

cell signaling and in the generation of amyloidogenic fragments. 

European Community 

3 years (2004-2007) 

Within the WP 1 Generation and Turnover, our proposal is aimed to the definition of the role 

that posttrasductional modifications of APP, in particular its phosphorylation, may exert on its 

pathophysiologycal function. We will investigate both the role of phosphorylated APP/CTFs in 

cell signaling, and/or in the generation of amyloidogenic fragments and plaques through the 

interaction with intracellular adaptors, as well as the effect that site-specific tyr-phosphorylation 

would have on the direct amyloidogenic pathway of APP. These studies, which are focused in a 

completely new aspect of APP activity, would lead to the identification of early markers of the 

neurodegenerative process which occurs in AD, and to the definition of a potentially innovative 

therapeutical approach based on the findings obtained. Our proposal is aimed at: 

Aim 1) To study directly in human brain the tyr-phosphorylation of APP and its CTFs and their 

possible coupling with intracellular adaptors such as ShcA, and to correlate these data with the 

presence of neuropathologycal hallmarks of the disease. The comparative analysis in non-AD 

control, AD and/or Down’s syndrome subjects at different ages (from fetal to adult life) will 

allow a deeper characterization of the molecular determinants during the progression of the 

disease. 

Aim 2) To study in transgenic mice carrying APP and PS1 mutations the tyr-phosphorylation 

of APP and CTFs, their interaction with ShcA-Grb2 adaptors and the influence that such 

interaction could have on plaque formation, astroglial response, neuronal death. The use of a 

Tg model will allow at a deeper comprehension of the possible correlation between APP 

phosphorylation, its cleavage, signaling activity and progressive presence of the typical 

hallmarks of the disease. Moreover, in this model is possible also to determine the effect that 

APP and PS1 mutations, which are linked with a human familial phenotype, would cause in the 

above mentioned parameters, and, in a second step, would allow the application of targeted 

therapeutical pharmacological approaches following also the indications raised from the in vitro 

studies (aim 3). 

Aim 3) To study in neuronal and/or glial cell cultures the molecular mechanisms which regulate 

APP phosphorylation, influence on cleavage by secretases, coupling with intracellular adaptors, 

and signaling activity. Using APP mutants, ShcA mutants, and APP KO cells we will define the 

influence that phosphorylation and interaction with ShcA would cause in signaling activity, cell 

proliferation and death, and in the generation of amyloidogenic fragments. Moreover, the study 

of the effect that proliferative and apoptotic stimuli may cause on those parameters and/or with 

the study of the kinases involved in the activation of APP cleavage and signaling, this would 

also lead to the identification of pharmacological target for a therapeutical intervention.

Nome Dr. Andrea Stracciari, dr.ssa Maria Guarino, prof. Fabio 

Contatti 

Cirignotta 

Istituto/Dipartimento Unità Operativa di Neurologia 








Azienda Ospedaliero-Universitaria S. Orsola-Malpighi, Bologna 

Dipartimento Scienze Neurologiche - Università di Bologna 

New treatment strategies (non pharmacologic therapeutic 

strategies in neurodegenerative diseases) 

Cognitive training in Parkinson’s disease 

Fondazione del Monte di Bologna e Ravenna 

24 mesi 

Background. Patients with Parkinson’s disease (PD) frequently 

present cognitive disturbances, which impair their autonomy and 

quality of life and are correlated with a significantly increased death 

rate.

Objectives. The primary objective is to assess improvements in 

quality of life in PD patients undergoing cognitive training. 

Secondary end-points will be changes in cognitive capacity, 

autonomy, mood, clinical judgement and burden to caregivers. 

Methods. This is a prospective randomized, 8-week, single blind, 2arm, 

multicenter, parallel-group study to evaluate the effect of task 

specific cognitive training on quality of life in patients with cognitive 

impairment, but without dementia (MMSE> 24), due to idiopathic 

PD. Fifty patients per arm will be randomized to detect a 10 point 

difference in the PDQ-39 overall score. 

Expected results. An improvement in quality of life obtained by 

enhancing cognitive functions, namely executive functions, is 

expected a the main result, being considered a clinically relevant 

outcome.

Nome Orazio Zanetti 

Contatti ozanetti@fatebenefratelli.it 

Istituto/Dipartimento IRCCS Centro San Giovanni di Dio Fatebenefratelli- Brescia 


Non invasive brain stimulation an integrated approach to neurorehabilitation in Alzheimer Disease 

Alzheimer’s Disease (AD) is a progressive disorder that impacts memory, language and several other 

cognitive functions. Given the limited effectiveness of pharmacological treatments, non-pharmacological 

interventions in AD have gained attention in recent years, and there are currently many different 

approaches under study, ranging from multi-strategy approaches to cognitive training (Cotelli et al 2006). 

Despite the potential therapeutic impact of the non-pharmacological approaches, the neural mechanisms 

underlying the beneficial effects of behavioral interventions remain largely unknown. Functional 

neuroimaging studies have shown that rehabilitation in patients with developmental and acquired 

cerebral damage may lead to functional cortical reorganization, a process mediated by activity-dependent 

plasticity mechanisms (Warburton et al 1999; Buckner et al 2004). These “plastic” mechanisms may also 

play a role in the aging brain and in Alzheimer Disease (Cotelli et al 2006). An increase in the activation of 

areas involved in memory, together with the recruitment of new areas during memory intensive tasks, has 

been confirmed using neuroimaging techniques in AD patients who underwent behavioral interventions 

(Cotelli et al 2008; Manenti et al 2008; Cotelli et al 2010 in press). 

Recent studies have reported enhanced performance on specific cognitive tasks in patients with several 

types of neurological disease, after receiving non invasive brain stimulation (BS), i.e., repetitive 

Transcranial Magnetic Stimulation (rTMS) or transcranial Current Stimulation (tCS) to specific cortical areas 

(Miniussi et al., 2008). 







New treatment strategies

Nome PATRIZIA BISIACCHI 

Contatti 049 8276587 

Istituto/Dipartimento DIPARTIMENTO DI PSICOLOGIA GENERALE 


Neurodegeneration occurring during normal ageing affects several cognitive processes, particularly 

memory, attention, executive functions. Within these functions there are specific aspects that are 

particularly susceptible to impairment due to unhealthy ageing, such as working memory, control 

mechanisms and sustained attention. These are every day cognitive functions that are a result of a 

sequence of complex neurological processes. The bio-signals (i.e. EEG, NIRS etc) recorded from a set of 

specific cognitive tasks have important roles in the assessment of these processes. Discovering features in 

signals of speech and visual processing, which are modalities that are must less open to subjective 

interpretation, would remove the subjective nature of some existing assessments. These modalities 

provide signals that can be collected remotely without significant amounts of resources and enable large 

research corpora of data, of great clinical significance, to be gathered effortlessly 

Specific focus of our researches is on the assessment of cognitive decline in older persons through the 

use of multimodal signal processing (i.e. behavioural, EEG, ERPs, fNIRS, Eye movements etc.) . 

Current clinical measures of cognitive status and general wellness of the elderly have several limitations. 

Most must be administered by trained professionals, the results of these measures can be affected by 

many external factors and the ratings can be affected by subjective scoring. Discharging elderly patients 

from hospital can often rely on a clinician’s subjective opinion about the patient’s ability to function in 

their own home. The possibility to utilize a remote multimodal assessment will bypass such difficulties. 


identificata 




- Imaging techniques and protocols 

- Standardization of diagnostic criteria and diagnostic 

instruments, harmonization and assessment tools 


- Ambient Assisted Living, Home automation, smart 

homes and domotics 

- Cognitive Ergonomics, ICT for elderly/Gerontechnology 

ELDERGAMES (Development of High Therapeutic Value IST-based 

Games for Monitoring and Improving the Quality of Life of Elderly 

People) 

PRINCIPAL INVESTIGATOR: PROF L. GAMBERINI 

Ente finanziatore EU FP6 

€ 100.373,00 

Durata progetto 01/09/06 - 28/02/09 

Abstract del progetto Motivating and amusing environments could help elderly 

people to preserve their cognitive functions and to improve their 

social life. Based on these premises, ELDERGAMES project


developed a mixed reality table-top solution on which users have to 

exploit and train their cognitive abilities through mind-challenging 

games in both collaborative and competitive settings 

SENIORCHANNEL (ICT based solutions for the advancement of 

social interaction of elderly people) 

PRINCIPAL INVESTIGATOR: PROF L. GAMBERINI 


EU- AAL/ MIUR 

€ 303.000,00 


Abstract del progetto Successful aging requires the involvement of elderly people 

in activities such as interacting and sharing their knowledge, 

opinions and aspirations with the wider community. 

SENIORCHANNEL project aims at supporting these activities 

through the development of a low-cost interactive IPTV and TV 

studio, through which elderly people will be able to create, interact 

and enjoy contents exploiting their creativity and knowledge. 


EARLY COGNITIVE AND ELECTROENCEPHALOGRAPHIC MARKERS OF 

NORMAL AND PATHOLOGICAL AGEING 

PRINCIPAL INVESTIGATOR: PROF P. BISIACCHI 


CARIPARO (Cassa di Risparmio di Padova e Rovigo) 

€ 45.000,00 

Durata progetto January 2009 – December 2011 

Abstract del progetto The main goal of the project is to investigate cognitive and 

electrophysiological alterations in healthy old individuals and in 

MCI patients, in order to early identify markers of pathological 

ageing. 



LIFESPAN CHANGES IN ELECTROPHYSIOLOGICAL PATTERNS 

ASSOCIATED WITH TEMPORAL DISCRIMINATION 


Bial Foundation 

€ 45.000,00 

Durata progetto January 2009 – December 2011 

Abstract del progetto The present project is aimed at clarifying the specific contribution 

of frontal and parietal brain regions in time processing, and their 

development and decline with age by means of new EEG and ERPs 

methodologies of signal analyses. Results might further have 

clinical application in diagnosis and rehabilitation of Parkinson’s 

disease and dementia. 


AN INTEGRATED APPROACH TO NEUROCOGNITIVE ASSESSMENT OF 

OLD PEOPLE 



FSE (Fondo Sociale Europeo) Regione Veneto 

€ 55.372,00






March 2009 – February 2010 

The present project, thanks to the integration of psychological, 

medical and bioengineering competencies, was aimed at 

developing systems of analysis that optimize the use of ERP/EEG 

methodology in the screening of executive functions in old people. 

IMPLEMENTATION OF NEW EEG SIGNAL ANALYSES FOR THE 

DETECTION OF SUBCLINICAL COGNITIVE IMPAIRMENT 


FSE REGIONE VENETO 

€ 49.418,18 

01/04/10 - 31/03/11 

Abstract del progetto The goal of the project is to create a new integrate approach to 

detect subclinical cognitive alterations in many pathologies. 


Cross Ages (Inter-generational learning. From diagnostic to impact 

evaluation) 

PRINCIPAL INVESTIGATOR: PROF R. DE BENI 


EU 

€ 126.649,00 

Durata progetto 01/12/07 - 30/11/09 

Abstract del progetto Promoting successful active ageing and social inclusion of older 

people in the local communities through the elaboration of an 

integrated lifelong learning model based on the intergenerational 

exchange.

Nome Vania Broccoli 

Contatti 

02 26434616 

broccoli.vania@hsr.it 

Istituto/Dipartimento Neuroscience Division 


Area di interesse identificata iPS technology offers also an unprecedented opportunity to recapitulate 

both normal and pathologic human tissue formation in vitro, thereby 

enabling disease investigation and drug development. 

PD might greatly benefit from an approach with iPS, since it is completely 

missing a valuable in vitro model of this pathology. In fact, up to now, 

primary skin fibroblasts and lymphocytes have been the only 

human cells available for investigating the pathogenetic mechanisms of 

PD. Although, they provided some insights on the disease, their 

completely different nature and metabolism prevents gaining information 

on the neuronal-specific degeneration processes. To overcome this 

limitation, disease affected neuronal cells 

would provide a completely new experimental system where to 

investigate the molecular roots of PD. 

This is even more important in PD, since also the animal models used for 

many years for this pathology have shown some important limitations 

failing to recapitulate the normal evolution of the pathological 

process observed in the patients. In fact, most of the studies have relied 

on mouse models of PD, which might be divided in genetic or 

symptomatic. Mutant animals for the PD-genes have permitted the 

investigation of specific processes associated to neuronal death such as 

oxidative stress and mitochondrial dysfunctions, but usually lack key 

features of the human disease as for instance selective degeneration of 

dopaminergic neurons. 

On the other hand, neurotoxins (6-hydroxydopamine, Rotenone, MPTP, 

Paraquate) have been widely used to efficiently induce dopaminergic 

neurodegeneration in animals, but again, some of the specific 

pathological signs of the human disease are not recapitulated as, for 

instance, the formation of Lewy bodies, the dynamics of neuronal death 

and a comparable development of the neurobehavioral deficits. 

For these reasons, the establishment of an iPS model system for PD might 

result particular significant. 

Indeed, there are some important features, which might enable this 

technology to be particular informative in PD research. First of all, some 

different procedures have been proved rather efficient in 

generating dopaminergic neurons starting from human ES and iPS cells. In 

particular, a treatment with FGF8 and Sonic Hedgehog (Shh) acts a strong 

enhancer of the in vitro dopaminergic commitment. These 

factors could be associated with different in vitro conditions such as coculture 

systems with stromal cells or SMAD inhibitors that strongly induce 

neuronal specification (Barberi et al., 2003; Chambers et al., 

2009). Moreover, the identification of genes whose mutations cause 

dopaminergic neuronal loss in a mainly cell-autonomous fashion allows to 

generate lines of iPS cells with a known genetic alteration and 

compare them to those derived from PD sporadic cases. 

Furthermore, we plan to use iPS-derived dopaminergic neurons as a 

source of transplantable cells in 6-hydroxidopamine-treated rats and 

mice. Neurons will be grafted in either the striatum or the substantia



nigra and their survival, integration and functional activity will be 

analyzed. Behavioral studies will assess whether transplanted cells will be 

able to rescue the neurological impairment in these animals. 

Modeling Parkinson’s disease by iPS technology: generation of human 

affected 

dopaminergic neurons and gene disease correction by site-specific 

integration 

Ente finanziatore ERANET-NEUON- EU 


Abstract del progetto Induced pluripotent stem (iPS) cells derived from somatic cells of patients 

represent an innovative tool for in vitro modeling of diseases and may 

provide a source for replacement therapies. In Parkinson disease (PD), 

the identification of the pathophysiological mechanisms behind this 

common neurodegenerative pathology has been particularly hampered 

by the lack of genuine in vitro models and animal models faithful to the 

human pathology. 

Exploiting new generations of viral vectors expressing the reprogramming 

(Oct4, Sox2 and Klf4) genes in fibroblasts from skin biopsies of the 

patients, we aim to establish faithful pluripotent iPS cell 

ines from sporadic and various monogenic forms of PD carrying 

mutations in either SNCA, PINK1, Parkin, LRRK2, DJ-1 or UCH-L1. In order 

to create an unprecedented in vitro culture system for PD affected human 

dopaminergic (DA) neurons, iPS cells will be differentiated in vitro 

through the Shh/Fgf8 stepwise induction protocol. Such patient-specific 

neurons will provide a system where we will investigate the mechanisms 

of genetic PD such as neuronal morphology, Lewy body formation, 

mitochondrial dysfunction, cell viability after oxidative and other 

stressors, interactions between different disease proteins, as well as 

electrophysiological deficits. For those experiments requiring a pure 

population of DA neurons, Pitx3-GFP iPS cell lines will be generated by 

means of BACtransgenesis enabling specific labelling of nigral DA 

neurons. Thus, pure populations of GFP+ dopaminergic neurons will be 

utilized for microarray based transcriptome profiling comparing PD versus 

control neuronal populations. These results promise to add precious 

information on both cellular and molecular pathological mechanisms 

leading ultimately to understand PD progression. 

Further, the establishment of human PD affected neurons will offer a 

remarkable cell platform for future systematic drug screenings. Finally, 

we will exploit an innovative strategy for site-specific integration of the 

therapeutic gene based on zinc finger nucleases mediated site-specific 

integration. This procedure will complement Parkin mutated iPS cells by 

targeting the therapeutic gene in a safe and transcriptionally competent 

genomic harbour. 

Overall, this project has the ambitious task to pioneer new approaches 

for iPS cell generation and combining them to develop safe gene 

correction strategies. These new procedures represent a big advantage 

for a broad functional characterization of iPS-derived dopaminergic 

neurons affected by PD. The successful achievements of our goals will 

disclose new opportunities for disease modelling, drug-screenings and 

cell replacements strategies in PD.

Nome Neural regeneration in the cerebellum: development of cell replacement 

strategies for the management of spinocerebellar ataxias. 

Contatti 

Tel. 

E mail 

Gian Giacomo Consalez, Ferdinando Rossi 

02 2643 4838 

giacomo.consalez@hsr.it, ferdinando.rossi@unito.it 

Istituto/Dipartimento Istituto Scientifico San Raffaele, Università degli Studi di Torino 


Optical And Biomolecular Methods For The Functional Investigation Of Neural Circuits In Vivo. 


rigenerazione in modelli di malattia neurodegenerativa 

identificata 



Neural regeneration in the cerebellum: development of cell replacement 

strategies for the management of spinocerebellar ataxias. 

Ente finanziatore Ataxia UK 


The project will use the postnatal and adult mouse cerebellum as a model system to set up cell 

replacement protocols to treat degenerative disorders of the central nervous system and, namely, 

spinocerebellar ataxias. 

In this project, we plan to: 

1. determine the ability of PC progenitors isolated from the embryonic cerebellum to proliferate in the 

host tissue and integrate into the wildtype and mutant cerebellar cortex 

2. analyze the spatio-temporal distribution of cues guiding PCs in their migration into the developing and 

adult cerebellar cortex 

3. manipulate the system to restore developmentally regulated cues guiding PC progenitor homing and 

connectivity 

The cerebellum is a highly plastic system whose development is completed after birth. Numerous 

observations show that exogenous PCs grafted to the cerebellum may integrate productively and establish 

proper synaptic connections with their afferents and post-synaptic targets. Several Mendelian cerebellar 

disorders, metabolic or neurodegenerative, feature a selective loss of PCs, fully warranting an attempt to 

develop cell replacement strategies. 

We plan to analyze the ontogenetic mechanisms that direct the integration of PC progenitors in the 

cerebellar cortex of wild type and spinocerebellar ataxia type 2 mice, and determine whether they are still 

available (or they can be re-activated) in the adult to promote repair. Our strategy for cerebellar 

regeneration will be driven by this analysis, in an attempt to exploit existing cues and to enhance those 

that are lost in time, eventually favoring the integration of exogenous precursors in the mature 

cerebellum. 

Protocols suited to promote the anatomical and functional integration of early PC progenitors into the wt 

or mutant mouse cerebellum.

Nome Federico Schena 

Contatti 

Federico.schena@univr.it 

Istituto/Dipartimento Dipartimento di Neuroscienze-Sezione Scienze Motorie Università di 

Verona & Centro di ricerca in Bioingegneria e Scienze Motorie, Rovereto 



identificata 






B. Physical activity programs for elderly 

subjects with different grades of cognitive 

impairment. 

Walking program for Alzheimer patients. 

Comune di Rovereto – Assessorato alle attività sociali 

Fondazione Mazzali - Mantova 

3 years 

Today, one of the main purposes of institutional long-term care is to offer 

well-organized therapy that also enhances the patients’ functional 

abilities. New integrated clinical approaches concerning physical exercise 

and Alzheimer’s disease (AD) have received attention in the scientific 

literature. Apart from individual rehabilitation, in the special care unit, 

patients rarely do physical activity alone; this phenomenon has been seen 

to cause a decline in the patients’ functional progress. 

Motor inactivity and asthenia become evident in the later stages of AD, 

including the ability to step, walk, and sit up independently. There is 

evidence, however, that specific physical activities can slow down this 

inexorable decline. For example, Fiatarone et al. demonstrated that in 

very old subjects high intensity resistance training and nutritional 

supplementation increased gait velocity, stair climbing power, 

spontaneous activity level, cross-sectional thigh area and maximal leg 

strength. Moreover, reports in the literature and recent guidelines 

suggest that the oldest-old can enhance cardiovascular function, 

flexibility, balance and strength through systematic exercise training. 

Most researchers in this field agree that also patients with Alzheimer’s 

disease can benefit from a targeted exercise program, because exercise 

has additional benefits for elderly dementia patients. [8-10] This notion 

has been proven in studies on AD patients; which have reported

numerous positive effects of physical exercise and walking programs: 

improvement in walking endurance, better urinary continence, enhanced 

communication, reduced depression and an increase in activities of daily 

life (ADLs). 

However in this institutional scenario, caregiver intervention and 

interaction with the medical staff and patients are rather difficult. In a 

nursing home, however, these important positive relationships were 

easier to manage. 

The aim of this program is to determinate whether an institution-based 

walking program carried out together with caregivers would reduce the 

functional, cognitive and physical decline of patients in the later stages of 

Alzheimer’s disease. The walking program will be compared with routine 

care in a randomized controlled clinical trial. We hypothesized that 

patients in the walking program would show improvements in physical 

capacity, cognitive performance and biomedical outcomes compared 

with those in the control care group.

Nome 

Prof. Enrico Granieri 

Contatti 

enrico.granieri@unife.it 

patrik.fazio@unife.it 

gino.granieri@unife.it 

Istituto/Dipartimento Sezione di Neurologia, Dipartimento di Discipline Medico-chirurgiche della 

Comunicazione e del Comportamento, Università di Ferrara 



identificata 




Promotion of Adapted Motor Activity among patients with Parkinson’s 

disease 


Fondazione Cassa di Risparmio di Cento, 

University of Ferrara 


Abstract del progetto An adapted motor activity could improve the pattern of body 

accelerometer and quality of life of patients with Parkinson’s disease. A 

pleasant, playful motor activity associated with music can give a motor 

performance improvement in terms of velocity and acceleration of the 

gait, stability in posture and better and rapid execution of movements. 

Objective of the study is to demonstrate with different technique and 

measures whether or not objective differences are detectable in subjects 

with Parkinson’s disease after an adapted motor activity based on 

emotional involvement through pleasant exercises, such as music and 

dance. For this study we developed a triaxial accelerometer that can be 

located in three different locations of the body that allow to measure the 

average accelerations in the body trunk. 

At least 100 patients suffering from Parkinson’s disease will be evaluated. 

The adapted motor activity session will be run twice a week, each of 

which lasted 60 minutes. They will be tested at entry of the study, after 4 

months of continuative motor activity and after 3 months following the 

end of the sessions by accelerometer test, disability scale (UPDRS III), 

quality of life questionnaire (SF-36), MMSE, tapping test. Measures of 

different domains with tests and objective measures with accelerometric 

techniques could improve our capability for the clinical monitoring of 

patients with movement disorders as Parkinson disease. In the same 

contest we aim to demonstrate the efficacy, in clinical terms, of an 

adapted motor activity based on emotional involvement through 

pleasant exercises with the support of music as a powerful cognitive and 

affective stimuli. 

(preliminary results of pilot studies has been shown at the Congresses of 

Italian Society of Neurology and LIMPE in 2008, 2009 and 2010, at the 

Congresses of Neurology in Moskow [Russia] and Belgrade [Serbia] in 

2010. Papers are submitted for publication in neurological journals).

Nome Dott.ssa Mariachiara Sensi 

Contatti 

mchiasen@gmail.com 

Istituto/Dipartimento U.O.Neurologia ,Dpartimento di Neuroscienze Riabilitazione, 

Ospedale S.Anna Ferrara 



identificata 







New treatment strategies (non pharmacologic 

therapeutic strategies in neurodegenerative diseases) with Deep brain 

Stimulation in dystonic and Parkinson Disease patients. 

None 

3 years 

To evaluate the long term response of Deep Brain Stimulations on motor 

and non motor symptoms in Parkinson disease and Dystonic Patients. 

The hypothesis is to test axial symptoms, dysartria, dysfagia, pain, both 

clinically and with non invasive imaging techniques .


F. Drug Pharmacogenetics

Nome Alberto Pilotto 

Contatti 

Phone: +39 0882 410271 

Fax: +39 0882 410271 

E-mail: alberto.pilotto@operapadrepio.it 

Istituto/Dipartimento Unità Operativa di Geriatria & Laboratorio di Gerontologia-Geriatria 

Dipartimento di Scienze Mediche 

IRCCS Casa Sollievo della Sofferenza 

Viale Cappuccini, 1 – 71013 San Giovanni Rotondo (FG) 


Sporadic Alzheimer’s disease (AD) is a progressive neurodegenerative disorder occurring predominantly in 

older age. The prevalence of AD rise from 20% after 75 years to 30% after 85 years, and about 5% of 

people aged 65 years or older have AD. With about 3 to 4 million people affected in the United States and 

about 350,000 new cases per year, AD is the most frequent cause of dementia in U.S. and in Western 

countries. Among the main cause leading to AD, the deficit of cholinergic system plays a major role. 

Accordingly, one of the most common therapy for the symptomatic treatment of AD is the block of 

acetylcholinesterase (AChE) by means of acetylcholinesterase inhibitors. The inhibition of AChE increases 

the concentration of acetylcholine (ACh) in the synaptic cleft, thus restoring the physiological effects of 

ACh within the central nervous system. Recent studies reported a significant benefits of 

acetylcholinesterase inhibitors vs placebo on cognitive function, activities of daily living, and behavior. 

These improvements, however, are not always detectable in clinical practice. Thus, it has been recently 

suggested that the detection of improvements on cognitive function, activities of daily living, and behavior 

may be obtained by the concomitant use of drugs acting on other pathogenetic AD mechanism. In 

particular the European Medical Agency (EMEA) recommend the use of the AChE inhibitors donepezil and 

rivastigmine in mild-to-moderate AD, and the addition of memantine in moderate-to-severe AD. 

Memantine block the glutamate receptor, limiting the uncontrolled entrance of Ca 2+ ion in the 

postsynaptic neuron, thus delaying the Ca 2+ -excess neurodegeneration. Most studies reported that 

interindividual differences in response to these drugs may be due to variability in drug metabolism related 

to behavioral, clinical, and genetic factors, mainly hereditary polymorphisms of drug-metabolizing and 

drug-transporting enzymes. Final objective of this proposal is to identify the genetic component 

underlying the response/non response to the most common drugs currently used in the treatment of AD, 

i.e. donepezil, rivastigmine and memantine, throughout the following specific objectives: 1) identification 

of significant difference in the distribution of the genotypes of the CYP2D6 gene polymorphisms among 

patients responders/non-responders to donepezil treatment; 2) identification of the specific DNA 

alteration producing modifications in the CYP2D6 enzyme activity; 3) confirmation of the modification of 

the enzyme activity by means of donepezil dosage in vivo; 4) identification of significant difference in the 

distribution of the genotypes of the acetylcholinesterase gene polymorphisms among patients 

responders/non-responders to rivastigmine treatment; 5) identification of the specific DNA alteration 

producing modifications in the acetylcholinesterase-mediated hydrolysis of rivastigmine; 6) confirmation 

of the modification of the enzyme activity by means of rivastigmine dosage in vivo; 7) identification of 

significant difference in the distribution of the genotypes of the OCT2 gene polymorphisms among 

patients responders/non-responders to memantine treatment; 8) identification of the specific DNA 

alteration producing modifications in the OCT2 transporter of memantine; 9) confirmation of the 

modification of the enzyme activity by means of memantine dosage in vivo. We expected that the 

identification of functional polymorphisms in the CYP2D6, AChE and OCT2 genes may influence the clinical 

efficacy of donepezil, rivastigmine and memantine in AD patients, and may be useful in identifying 

subgroups of AD patients with different clinical response to treatment. Criteria and indicators to verify 

results are: 1) the number of patients enrolled in the study in the first 6 months; 2) the number of 

genotypes investigated at the end of the 1 th year of the project; 3) the number of specific gene alteration 

influencing the enzyme activity at the 2 nd year of the project; 4) the quantitative data obtained from 

plasma drug dosage at the end of the 2 nd year of the project. This project is coherent with the ministerial

guidelines for the I.R.C.C.S. and well integrate the clinical practice in neurology. 


identificata 






Pharmacogenetics of drugs for Alzheimer’s Disease 

Effect of weight loss on metabolic, functional, cognitive status and 

biological markers of longevity in obese frail elderly 

Ministero della Salute 

Two years 

Obesity, and in particular abdominal obesity causes serious metabolic and 

medical complications and impairs quality of life. Moreover, in elderly 

persons, obesity can lead to frailty by promoting chronic inflammation 

and by exacerbating the decline in strength, endurance, balance and 

mobility associated with aging and physical inactivity. Weight loss, 

induced by calorie restriction and/or physical exercise, simultaneously 

improves multiple metabolic risk factors for cardiovascular disease and 

other medical abnormalities associated with obesity, and reduce 

morbidity and mortality. However, the appropriate treatment for obesity 

in elderly persons is controversial because of the potential harmful 

effects of weight loss on bone and muscle mass. It has been reported that 

weight loss can also modify biological pathways at the cellular level. 

Insulin and of insulin-like growth factor-1 (IGF-1) have been involved in 

cellular caloric equilibrium and other mechanisms, i.e. oxidative stress 

and inflammation. Thus genetic polymorphisms in genes involved in these 

metabolic pathways such as apolipoprotein E (APOE), sterol regulatory 

element-binding protein cleavage-activating protein (SCAP), peroxisome 

proliferators-activated receptor gamma-2 (PPRγ), α-2B-adrenergic 

receptor (ADRA2B), acyl-CoA synthetase 5 (ACSL5) and interleukin-6 (IL-6) 

may influence the effect of weight loss on biological mechanisms and 

possibly on functional and cognitive status in the frail elderly. It is possible 

that weight loss in obese elderly persons can be beneficial by improving 

metabolic, functional and cognitive status, or harmful by causing a 

decrease in muscle and bone mass. However, the clinical and 

physiological effects of weight loss in obese elderly subjects have never 

been carefully evaluated. Thus, appropriate treatment for this rapidly 

increasing segment of the elderly population remains controversial. The 

purpose of this project is to advance our scientific knowledge by 

determining the effects of weight loss on metabolic, functional and 

cognitive status in obese elderly subjects. Moreover, we will evaluate the 

role of weight loss on biological markers of metabolic health and 

longevity, i.e. serum level of Insulin, insulin-like growth factor-1 (IGF-1), 

transforming growth factor-β (TGF-β), tumor-necrosis factor-α (TNF-α), 

interleukin-1 (IL-1) and C-Reactive Protein (CRP), and their relationships 

with genetic polymorphisms in APOE, SCAP, PPR-γ, ADRA2B, ACSL5 and IL- 

6 genes. As biomarker of longevity we will also evaluate the accumulation 

of mitochondrial (mt) DNA mutation through the analysis of the D310 

mononucleotide repeat of mtDNA.





Smart Home for Elderly People (HOPE) 

Ministero della salute/Ministero dell’Università e della Ricerca Scientifica 

Two years 

Since Europe’s and worldwide population grows older, the need for care 

is growing as well as people spend more money to get it. The solution 

Smart Home for Elderly People (HOPE) promises to reduce the need for 

carers, improve the life of older people and cut the cost of assistance. The 

main objective of the “Hope” project is to produce an Integrated 

Computer Technology (ICT) solution that will help the elderly people, 

specifically those that suffer with the Alzheimer s disease (AD), achieve a 

richer and more independent lifestyle. Dementia causes long and 

oppressive suffering to patients and their relatives and imposes 

enormous costs on society. About 25 million people suffered from 

dementia in recent years. As a 4-fold increase of this number is expected 

by 2050, dementia is one main health issue of the next decades. AD 

covers 50-70% of all dementia cases, no cure exists, and effective and 

reliable early diagnostic techniques are lacking. Early diagnosis and 

progress monitoring of AD is a central part of treatment until future drugs 

and prevention strategies become available. Elderly people with AD 

spend most of their time at home. The “HOPE” solution consists of an 

integrated, smart platform that will enable the elderly people with AD to 

use innovative technology for a more independent life, easy access to 

information, monitor their health, and serve as a source of inspiration for 

users as well as for people working with assistive devices. Moreover, it 

will enable them to perform by themselves activities they were not able 

to do before and which are important for their daily personal life. The 

main advantage of the proposed system is that it provides a basis for 

integrating services for the elderly population while they are at home. 

HOPE is a budgeted solution that will be installed at the elderly people’s 

homes, and will provide services for (a) life-long, self organized, 

appropriate educational environment and access to information, (b) care 

management and health support, (c) self monitoring and decision making. 

Optionally, the user will be able to activate software which automatically 

establishes the necessary interactive, triple-play connection for receiving 

tele-help and tele-assistance from specialized service providers, doctors 

or other medical personnel. The proposed application will be intelligent 

enough to offer safety in terms of controlling efficiently the home 

environment, economy in terms of controlling and decreasing the need 

for external help, and convenience in terms of adjusting the operation of 

connected sub-systems to achieve the best possible user experience.


G. Clinical studies

Nome Prof. Carlo Caltagirone 

Contatti 

Tel. 

E mail 

Via Ardeatina, 306 – 00179 Rome 

06 51501409 


Istituto/Dipartimento IRCCS Fondazione Santa Lucia – Laboratory of Clinical and Behavioral 

Neurology. 



identificata 






Neurocognition in Alzheimer Disease 

Mortality of elderly using conventional and non conventional 

antipsychotic drugs for the behavioural and psychological symptoms of 

dementia and for other psychical disorders. 

AIFA 

2007-2010 

Background. Antipsychotic drugs are used to control the 

behavioral and psychological disturbances that often occur in 

elderly, particularly in those who suffer from dementia. Nonconventional 

drugs are perceived by clinicians to be better tolerated 

than conventional ones and for this reason are more frequently 

adopted for frail and elderly patients. However, reports from 

ongoing randomized clinical trials on risperidone and olanzapine 

(two non conventional antipsychotic drugs) in elderly with dementia, 

signaled an increased mortality and incidence of cerebrovascular 

events in patients assigned to active drug with respect to placebo. 

After this initial report, a formal meta-analysis of fifteen randomized 

placebo-controlled trials, (3,353 patients randomized to 

nonconventional antipsychotic drugs and 1,757 randomized to 

placebo) confirmed the existence of a small increase in mortality in 

those patients who had been assigned to active drugs. For this 

reason, in European and extra European countries, the health 

authorities introduced some limitations to the use of these non 

conventional drugs. When a drug treatment for behavioral 

disturbances becomes necessary, these limitative measures favor 

the use of conventional drugs but it is not known whether 

conventional antipsychotics are safer than conventional ones. 

Objectives. To assess, in a large population of elderly 

residents in different Italian Regions (Piemonte, Emilia, 

Marche) and in the metropolitan area of Milan the overall 

and cause specific mortality associated with the use of

conventional and non conventional antipsychotic drugs. 

Methods. Using the informative systems of the regional health 

authorities and of the health district of Milan we will compose a 

population cohort formed by all the persons of 60 years or older 

who were prescribed an antipsychotic drug for the first time from 

January 2001. It is expected that this cohort might be of over 30000 

elderly persons. For each included person the last date of existence 

in life or the date of death will be retrieved. Mortality will be 

evaluated from first assumption of antipsychotic drug by type of 

drug used (conventional, non conventional) and by great groups of 

causes of death (tumors, cardio vascular diseases and other 

causes). Mortality will be separately assessed for elderly users of 

antipsychotic and antidementia drugs. 

Expected results. This retrospective analysis of survival on an 

extraordinarily large cohort will give further evidences about the 

potential role of treatment with antipsychotic drugs in survival of 

elderly with and without dementia.


A . Human –epidemiology/risk factors


Contact person: Gabbianelli Rosita or Cinzia Nasuti 

School of Pharmacy, University of Camerino, 

Date: 29/01/2010 


The main focus of our research is the development of a progressive neurodegenerative PD rat model. The 

research is based upon a multidisciplinary and integrated consortium bringing together field based 

researchers (genetic, molecular biology, biochemistry, toxicology and chemistry), from 1 Asian and 7 

European countries, and 1 SME (University Medical Center Groningen Netherlands; Institut für Organische 

Chemie der RWTH, Germany; Biological Research Center of the Hungarian Academy of Sciences, Hungary; 

Instituto de Tecnologia Química e Biológica, Portugal; Departamento de Engenharia Química ISEP, Portugal; 

Technological Educational Institute of Athens, Greece; Alagappa University, India; SME, Chema Diagnostica 

di Marco Fiore, Italy). 

Environmental contaminants accumulated through the food chain can perturb normal physiologic 

processes leading to the initiation and progression of the Parkinson’s disease (PD). The consequences 

deriving from early life exposure to these environmental factors can influence variability of life span and 

the health profile of the elderly in the human population. 

PD neuropathology is characterized by the degeneration of the nigrostriatal dopaminergic pathway. It is the 

second most common neurodegenerative disorder, affecting 1-2 % of the population over the age of 50. PD 

prevalence studies show that the number of individuals with PD over age 50 (4.1-4.6 millions) will double 

by 2030 in Western European nations (8.7-9.3 million). Moreover the burden of PD will also shift from more 

industrialized nations to developing Asian nations (India, China etc.) where survival of individuals is likely 

expected to grow with improving economic conditions and health care. 

Environmental factors such as lifestyle, diet, pesticides, metals and solvents, seem to be involved in the 

development of neurodegeneration by causing epigenetic modification in people with PD over 50 years old. 

Pesticides are one of the most important risk factors of epigenetic changes that affect the development of 

PD. Permethrin (PERM) is a pesticide mainly used in agriculture, in healthcare, in industrial and domestic 

settings, and in public health services. It is employed in tropical areas to prevent mosquito-borne disease. 

Several studies find that pyrethroids persist in house dust in significant concentrations for months after 

they are applied for pest control operation and specifically, their metabolites were found in greater than 50 

% in the urine of the subjects tested (Riederer et al., 2008). The potential consequences of pesticide 

exposure are greater for children than for adults. Children have higher risk of pesticide exposure due to 

behaviours such as mouthing items and playing on floors. Nevertheless, the primary exposure pathway of 

pyrethroids in humans is thought to be through dietary intake of vegetables, fruits and cereals. It is a 

consequence of pesticide residues that has accumulated in soil and groundwater that is re-cycled for 

irrigation of crops.

Given the complexity of the many factors to which the populations described in the PD epidemiological 

studies have been exposed, the criteria to identify candidate substances as causative factors of PD have to 

follow these points: 1) effects on the striatal dopaminergic system (decrease in dopamine level/increase of 

its metabolites, decreased in dopamine transporter and enzyme tyrosine hydroxylase activity; 2) damages 

on substantia nigra and striatum; 3) mechanistic effects (e.g. on oxidative stress, mitochondrial 

dysfunction/complex I inhibition, and alpha-synuclein aggregation). Our previous studies show that 

permethrin insecticide, belonging to the pyrethroids family, could be the ideal candidate to satisfy the main 

criteria suggested above. Firstly, mitochondria complex I activity was inhibited in striatum cells incubated 

with PERM (paper submitted to Neuroscience). Secondly, in vivo studies on pup rats treated with PERM 

during brain development (from 6 th to 15 th day of life) showed an imbalance in the dopaminergic system 

(Nasuti et al., 2007). High homovanillic acid levels and reduced dopamine concentrations were measured in 

the striatum of 35 day old rats treated. The period of treatment corresponding to neurodevelopment 

events (synaptogenesis) in rat brain, occurs in the third trimester of pregnancy in humans. Increased 

oxidative stress in striatum (glutathione depletion, increase of protein oxidation), blood cells (decrease of 

glutathione peroxidase, increase of lipid peroxidation, unbalance of monocytes and neuthrophils 

respiratory burst), and behavioural modifications were measured (Nasuti et al., 2007; Gabbianelli et al., 

2009, a; Gabbianelli et al., 2009, b). Thirdly, markers related to oxidative stress, such as DNA damage, 

changes in plasma membrane fluidity and antioxidant enzyme activities were observed in both striatum 

and blood cells of adult rats treated with PERM for two months (Nasuti et al.,2003, 2008; Gabbianelli et al., 

2002, 2004; paper submitted to Neuroscience). 

Based on assumptions described above we suggest the followed strategic issue: Pathogenic mechanisms, 

early diagnosis and prevention of Parkinson’s disease (PD) following pesticide intake. 

The research aims to capitalize on knowledge acquired in the field of pesticide effects on 

neurodegenerative disease starting from developmental to old age. Studies of the role of epigenetic 

mechanisms are a research priority. The studies take into account new approaches for early diagnosis of PD 

and strategies able to prevent the development of the neurodegeneration. 

Five major objectives to reach: 

1) Define the role of the early-life pesticide intake as a causative environmental agent for the development 

of PD; 

2) Define the correlation between pesticide and epigenetic mechanisms in regulating transcription of 

genes; 

3) Identify the specific peripheral markers related to early diagnosis of PD; 

4) Evaluate the risk following pesticide food intake and translate the outcome to human population; 

5) Prevention of PD onset and/or progression by three strategies: reduction of pesticide residues in the 

environment through bioremediation, screening of compounds for neuroprotective therapy and 

development of diagnostic kits to monitor peripheral markers related to the early PD diagnosis.


Contact person: Micaela Caserta (CNR) 

Date: January 28 th , 2010 


Increase in longevity does not correlate with an increase in disease-free life expectancy. The molecular 

mechanisms at the basis of this lack of correlation are still matter of debate and require research 

investment. 

Cellular senescence has long been used as a cellular model for understanding mechanisms underlying the 

ageing process. Compelling evidence obtained in recent years demonstrate that DNA damage is a common 

mediator for both replicative senescence, which is triggered by telomere shortening, and premature 

cellular senescence induced by various stressors such as oncogenic stress and oxidative stress. Extensive 

observations suggest that DNA damage accumulates with age and that this may be due to an increase in 

production of reactive oxygen species (ROS) and a decline in DNA repair capacity. Genomic instability is 

therefore thought to play a causative role in the ageing process. 

Our working hypothesis is based on the possibility that chronic exposure to environmental factors causing 

oxidative stress during life, including pollutants but also noxious nutrients such as alcohol, could actually 

reduce the ability of relevant genes, implicated in the process of protecting the cell from protein and DNA 

damage, to respond properly. If, at the same time, inhibition of programmed cell death (apoptosis) occurs, 

cellular damage starts to accumulate. 

We will develop experimental strategies to analyse the mechanisms regulating the delicate balance 

between rescue and apoptotic pathways in two model systems, human monocytic cells and yeast S. 

cerevisiae, with the specific aim to define molecular targets of pharmacological intervention. 


Human cells 

The major goal is to understand the molecular mechanisms by which environmental insults, causing 

oxidative stress in several cell types and tissues, alter the expression of genes involved in counterbalancing 

the damaging effects, like those coding for inducible heat shock proteins, detoxifying enzymes, proteins 

involved in DNA repair processes as well as factors controlling programmed cell death. 

One of the experimental systems that will be employed consist of human monocytic cell lines, that will be 

subjected to various stress agents (mutagenic stress, ethanol, heat shock, and oxidative stress) either as 

single treatment or as repeated cycles of treatment. The parameters that will be evaluated are: 

- mRNA level of selected genes (Hsp70, Hsp90, TRAP1, ERCC1, TDG, MCL1, DAD1); 

- chromatin remodelling and histone modifications occurring at their promoters;

- induction of apoptosis/necrosis; 

- epigenetic modifications and activity of PARP-1 and SIRT1, two key regulators of the balance between life 

and death of stressed cells. 

The subsequent step will be to evaluate some of these parameters (those requiring a limited number of 

cells) on primary blood monocytic cells. 

S. cerevisiae 

Saccharomyces cerevisiae has played an important role as a model system to understand the biochemistry 

and molecular biology of mammalian cells. The genetic tools available and the short life span have also 

made S. cerevisiae a powerful system to study aging. The yeast chronological life span (CLS) is a measure of 

the survival of a non-dividing population of cells, and thus can model aging of mammalian non-dividing cells 

but also of higher eukaryotic organisms. The parallel description of the pro-aging role of homologs of Akt, 

S6 kinase, adenylate cyclase, and Tor in yeast and in higher eukaryotes, suggests that findings in S. 

cerevisiae will be valuable to understand human aging and diseases. Moreover, the similarities between 

mitochondria and age-dependent mitochondrial damage in yeast and mammalian cells indicate that S. 

cerevisiae is a valuable model to study mitochondrial dysfunction and diseases involving this organelle. 

Here, we propose to study the relevance of epigenetic modifications due to the yeast sirtuins (Sir2, HST1-4) 

at relevant aging loci such as telomeres and ribosomal DNA genes. Analyses will be carried out in condition 

of oxidative stress and at different times of chronological life. The results obtained will be then compared 

with parallel treatments in human cells were the sirtuin set (seven members in human) could act in a very 

similar way. 

III) Impact 

The comprehension of the basic mechanisms underlying cell response to environmental cues, alongside 

with the increase of knowledge on human biological variation in health and disease, will allow to identify 

specific pathways and genes whose activity needs to be strengthened or reduced by pharmacological 

intervention.

Nome Valerio Carelli, MD, PhD 

Contatti 

Valerio.carelli@unibo.it 

Istituto/Dipartimento Department of Neurological Sciences, University of Bologna 


MELANOPSIN RETINAL GANGLION CELLS AND CIRCADIAN RHYTHMS IN PARKINSON AND 

ALZHEIMER DISEASE 


identificata 






Biomarkers (chronobiology and neurodegeneration) 

MELANOPSIN RETINAL GANGLION CELLS AND CIRCADIAN RHYTHMS IN 

PARKINSON AND ALZHEIMER DISEASE 

none 

2 years 

Background: 

Circadian rhythm abnormalities have been described in aging and 

neurodegenerative disorders such as Parkinson and Alzheimer disease 

(1,2). In particular, the “sundowning” (i.e. the appearance of 

psychomotor agitation during the evening-night) described in Alzheimer 

patients, has been related to an abnormal circadian rhythm of body 

temperature (3). In the last years many retinal abnormalities (mainly 

contrast sensitivity but also colour vision defects and others) and the 

presence of optic neuropathy have been documented in both Alzheimer 

and Parkinson disease (4-6). Moreover, in both diseases bright light

therapy has been reported to be effective (7,8). Nevertheless, no data are 

available on the possible relationship between visual dysfunction and 

circadian rhythm abnormalities in these disorders. Their possible link 

comes from the recent discovery of a new class of photoreceptors, the 

melanopsin RGCs (mRGCs) (9). Melanopsin retinal ganglion cells (mRGCs) 

are a new class of photoreceptors playing a crucial role in non-image 

forming visual functions such as entrainment of circadian rhythms to light 

and dark cycle, pupillary light reflex and regulation of synthesis of 

melatonin during the night (10). These cells represent about 1% of RGCs 

and originate the retinohypothalamic tract (RHT) (11). Furthermore, In 

the retina of patients with Parkinson disease a depletion of dopamine 

levels has been documented. The amount of this reduction is strictly 

influenced by the time of last levodopa assumption (12). It is of note that 

dopaminergic amacrine cells interact with mRGCs and regulate 

melanopsin gene expression (13-15). 

In collaboration with Professor Alfredo Sadun (Doheny Eye Institute, USC, 

Los Angeles, California) we recently demonstrated that mRGCs are 

partially spared in mitochondrial optic neuropathies, characterized by 

selective loss of RGCs (i.e. LHON and DOA). Our study also showed that 

mRGCs are lost after age eighty in normal subjects (16,17). 

Rationale: 

Based on the current knowledge we hypothesize that the abnormalities 

of circadian rhythms documented in Alzheimer and Parkinson disease 

may be related to the presence of optic neuropathy and involvement of 

the mRGCs system. 

Aims: 

The aims of this project are: 

- to verify the presence of rest-activity rhythms by means of actigraphic 

recordings in Parkinson and Alzheimer patients 

- to verify the presence of optic neuropathy by measurement of nerve 

fiber layer thickness, as evaluated with Optical Coherence Tomography 

- to verify and quantify the presence of optic neuropathy and the 

involvement of mRGCs in post-mortem retinas of Alzheimer and 

Parkinson patients 

- to verify the presence of dopamine depletion in Parkinson retina 

- to correlate the presence of optic neuropathy and circadian rhythm 

abnormalities in patients with Alzheimer and Parkinson disease. 

References: 

1. Wu YH, Swaab DF. Disturbance and strategies for reactivation of 

the circadian rhythm system in aging and Alzheimer's disease. 

Sleep Med. 2007 Sep;8(6):623-36. 

2. Willis GL. Parkinson's disease as a neuroendocrine disorder of 

circadian function: dopamine-melatonin imbalance and the visual 

system in the genesis and progression of the degenerative

process. Rev Neurosci. 2008;19(4-5):245-316. 

3. Volicer L, et al. Sundowning and circadian rhythms in Alzheimer's 

disease. Am J Psychiatry. 2001 May;158(5):704-11. 

4. Hinton DR, et al. Optic-nerve degeneration in Alzheimer's 

disease. N Engl J Med. 1986 Aug 21;315(8):485-7. 

5. Sadun AA, Bassi CJ. Optic nerve damage in Alzheimer's disease. 

Ophthalmology. 1990 Jan;97(1):9-17. 

6. Archibald NK, et al. The retina in Parkinson's disease. Brain. 2009 

May;132(Pt 5):1128-45. 

7. Paus S, et al. Bright light therapy in Parkinson's disease: a pilot 

study. Mov Disord. 2007; 22(10):1495-8. 

8. Dowling GA, et al. Effect of morning bright light treatment for 

rest-activity disruption in institutionalized patients with severe 

Alzheimer's disease. Int Psychogeriatr. 2005; 17(2):221-36. 

9. Berson DM, et al. Phototransduction by retinal ganglion cells that 

set the circadian clock. Science. 2002 Feb 8;295(5557):1070-3. 

10. Hattar S, et al. Melanopsin-containing retinal ganglion cells: 

architecture, projections, and intrinsic photosensitivity. Science. 

2002 Feb 8;295(5557):1065-70. 

11. Hannibal J, Fahrenkrug J. Neuronal input pathways to the brain's 

biological clock and their functional significance. Adv Anat 

Embryol Cell Biol. 2006;182:1-71. 

12. Harnois C, Di Paolo T. Decreased Dopamine in the Retinas of 

Patients with Parkinson's Disease. Investigative Ophthalmology & 

Visual Science, 1990; 3 1(11) 

13. Vugler AA, et al. Dopamine neurones form a discrete plexus with 

melanopsin cells in normal and degenerating retina. Exp Neurol. 

2007 May; 205(1):26-35. 

14. Sakamoto K, et al. Dopamine regulates melanopsin mRNA 

expression in intrinsically photosensitive retinal ganglion cells. Eur 

J Neurosci. 2005 Dec;22(12):3129-36. 

15. Zhang DQ, et al. Wong KY, Sollars PJ, Berson DM, Pickard GE, 

McMahon DG. Intraretinal signaling by ganglion cell 

photoreceptors to dopaminergic amacrine neurons. Proc Natl 

Acad Sci U S A. 2008 Sep 16;105(37):14181-6. 

16. La Morgia C et al. Melanopsin-containting retinal ganglion cells 

and circadian phototransduction are spared by 

Neurodegeneration in Mitochondrial Optic Neuropathies. 

Neurology March 17, 2009 Suppl 3 A183 (P04.075) 

17. La Morgia C et al. Light-induced melatonin suppression in 

Mitochondrial Optic Neuropathies. Arvo Annual Meeting, May 3-7

2009 Fort Lauderdale, Florida N. 5663

Nome Prof. Roberto Lucchini 

Contatti Tel +39030 3996604 Fax +39030 3996080 Email: lucchini@med.unibs.it 

Istituto/Dipartimento Dipartimento di Medicina Sperimentale e Applicata, Sezione di Medicina del 

Lavoro, Università di Brescia, P.le Spedali Civili 1, 25123 Brescia 



identificata 


1. Titolo progetto 




2.Titolo progetto 




3.Titolo progetto 




Gene-metal interaction as a determinant of neurodegenerative diseases 

PHIME - Public health impact of long-term, low-level mixed 

element exposure in susceptible population strata 

EU 6th Frame Program (www.phime.org) 

2006-2011 

PHIME: Gene-environment interaction on manganese induced 

parkinsonism are studied in children and elderly resident in areas with 

environmental exposure to heavy metals 

Epigenomics Studies of Toxic Hazards in Environmental health Research 

CARIPLO Foundation, Italy 

2008-2011 

This is an interdisciplinary program to conduct in-vitro and human 

investigations to evaluate the role of DNA methylation and histone 

modifications in mediating the health effects of metals and other 

environmental exposures 

PROFILO BIOLOGICO E GENETICO DELLA DISFUNZIONE DEI METALLI 

NELLA MALATTIA DI ALZHEIMER E NEL ‘MILD COGNITIVE IMPAIRMENT’ 

Ministero del Lavoro della Salute e delle Politiche Sociali 

2009-2011 

Several cross-sectional as well as cohort studies corroborate the concept 

of metal implication in Alzheimer’s disease (AD), including clinical 

observations suggesting that higher-than normal levels of copper (Squitti 

et al, Neurol 2002) and in particular ‘free’ copper – the quantity of serum 

copper which is not bound to ceruloplasmin (Squitti et al, Neurol 2005, 

2006) – can distinguish AD from healthy controls. Here we tested the 

hypothesis that this is the case also for the classification of healthy elderly 

subjects vs. Mild Cognitive Impairment (MCI) which represents an 

intermediate cognitive state between ‘normalcy’ and full dementia.

ABSTRACT 

Exposure to heavy metals can induce neurodegenerative effects in humans (Zatta et al., 2003). Various 

metals like aluminium, manganese, iron, copper, zinc are likely to play important roles in the pathogenesis 

of neurodegenerative illnesses such as Alzheimer’s Dementia (AD) and Parkinson’s Disease (PD). In 

particular for AD - the most common form of dementia in the elderly -a consensus has been recently 

established in the Metal Hypothesis of AD (Bush and Tanzi 2008). This hypothesis suggests that the 

interaction of Abeta – the major component of the senile plaques - with specific metals - iron, zinc and 

especially Cu - can drive Abeta pathogenicity and AD development and progression. Numerous clinical 

studies, including some from our group (Squitti et al, 2002, 2006, 2009) support this hypothesis. In 

particular, we have identified in AD patients a serum-level increase in the Cu quantity that does not bind to 

ceruloplasmin (free Cu) (Squitti et al, 2005), which correlates with the typical deficits, cerebrospinal 

markers (Squitti et al, 2006) and with a worse prognosis of the disease (Squitti et 2009a). The interaction 

between environmental exposure with genetic susceptibility is suspected as an important determinant of 

these chronic diseases as well (Wright and Baccarelli 2010; Baccarelli e Bollati, 2009). In fact, if on one hand 

metabolic dysfunction could have an effect on metal disbalance – especially at the liver lever, being the 

liver the controlling organ of metal homeostasis, on the other hand epigenetic factors have been 

highlighted in conjunction with oxidative stress mechanism to be triggered by exposure to neurotoxicants 

like heavy metals (Zawia et al., 2009). In particular, the effects of metal exposures in early life and 

throughout the developmental stages could account for clinical manifestations in older age. Our group has 

produced seminal contributions showing that exposure to metals (Wright and Baccarelli 2010; Tarantini 

and Baccarelli 2009) modifies global and gene-specific methylation states and that these changes can be 

detected in blood DNA. Epidemiological assessment needs accurate reconstruction of long-term and 

lifetime exposure, possibly with exposure biomarkers able to reflect long-term cumulative mechanism of 

neurotoxicity. Our group has developed various research projects in these areas, covering different aspects 

of metal exposure as determinant of AD (Papaleo et al., 2004) and PD (Lucchini et al., 2007; Squitti et al., 

2009b). The understanding of exposure related determinants have important implication in terms of 

possible reduction of the epidemiological impacts of neurodegenerative diseases, through preventive 

intervention. The line of research we are interested in focuses on an integration of all the aspects – 

exposure-related as well as the metabolic ones - of the metal implication in neuro-degenerative disorders. 

In particular, we will explore the genetic and epigenetic influence on AD and PD patients exposed to heavy 

metals in the highly industrialized province of Brescia, and the hypothesis that gene controlling metal 

metabolism – such as the Wilson’s disease ATP7B gene - which controls the free copper levels in the body, 

the hemochromatosis HFE gene and Transferrin gene - involved in the regulation of body iron - arbour 

susceptibility loci for late-onset AD as well . Moreover, to provide proof-of-concept that a decoppering 

approach has a beneficial effect on AD progression, we are currently starting a large randomized controlled 

clinical trial based on the concept that metal complexing or ligand agents properly tune the redistribution 

of metals in the body. The influence of genes like parkin and PARK9 will also be assessed in relation to 

exposure to neurotoxic metals like manganese. Assessment of cumulative exposure (Sinha and Mark, 2005) 

to metals will be based on geospatial approach and will be applied to AD and PD patients and age-sex 

matched controls from geographical areas with different environmental exposures to heavy metals.

REFERENCES 

Baccarelli A, Bollati V. Epigenetics and Environmental Chemicals. Curr Opin Pediatr 2009; 21: 243-51. 

Lucchini R, Albini E, Benedetti L, Borghesi S, Coccaglio R, Malara E, Parrinello G, Garattini S, Resola S, Alessio 

L, High Prevalence Of Parkinsonian Disorders Associated To Manganese Exposure In The Vicinities Of 

Ferroalloy Industries. Am J Ind Med 2007; 50: 11: 788-800 

Papaleo B, Alessio, Apostoli P, Battista G, Benedetti F, Lucchini R, Pasqualetti P. Alzheimer disease: study of 

occupational risk factors [in Italian]. G Ital Med Lav Ergon 2004;26(4):310-312 

Bush AI, Tanzi RE. Therapeutics for Alzheimer's disease based on the metal hypothesis. Neurotherapeutics. 

2008 Jul;5(3):421-32. Review 

Squitti R, Lupoi D, Pasqualetti P, Dal Forno G, Vernieri F, Chiovenda P, Rossi L, Cortesi M, Cassetta E, Rossini 

PM. Elevation of serum copper levels in Alzheimer's disease. Neurology. 2002 Oct 22;59(8):1153-61.. 

Squitti R, Pasqualetti P, Dal Forno G, Moffa F, Cassetta E, Lupoi D, Vernieri F, Rossi L, Baldassini M, Rossini 

PM. Excess of serum copper not related to ceruloplasmin in Alzheimer disease. Neurology. 2005 Mar 

22;64(6):1040-6. 

Squitti R, Barbati G, Rossi L, Ventriglia M, Dal Forno G, Cesaretti S, Moffa F, Caridi I, Cassetta E, Pasqualetti 

P, Calabrese L, Lupoi D, Rossini PM. Excess of nonceruloplasmin serum copper in AD correlates with 

MMSE, CSF [beta]-amyloid, and h-tau. Neurology. 2006 Jul 11;67(1):76-82. 

Squitti R, Bressi F, Pasqualetti P, Bonomini C, Ghidoni R, Binetti G, Cassetta E, Moffa F, Ventriglia M, Vernieri 

F, Rossini PM. Longitudinal prognostic value of serum "free" copper in patients with Alzheimer disease. 

Neurology. 2009a 6;72(1):50-5. 

Squitti R, Gorgone G, Panetta V, Lucchini R, Bucossi S, Albini E, Alessio L, Alberici A, Melgari JM, Benussi L, 

Binetti G, Rossini PM, Draicchio F. Implications of metal exposure and liver function in Parkinsonian 

patients resident in the vicinities of ferroalloy plants. J Neural Transm. 2009b Oct;116(10):1281-7 

Tarantini L, Bonzini M, Apostoli P, Pegoraro V, Bollati V, Marinelli B, Cantone L, Rizzo G, Hou LF, Schwartz J, 

Bertazzi PA, Baccarelli A. Effects of Particulate Matter on Genomic DNA Methylation Content and iNOS 

Promoter Methylation. Environ Health Perspect 2009; 117: 217-22. 

Wright RO, Schwartz J, Wright RJ, Bollati V, Tarantini L, Park SK, Hu H, Sparrow D, Vokonas P, Baccarelli A. 

Biomarkers of Lead Exposure and DNA Methylation within Retrotransposons. Environ Health Perspect, 

online ahead of press 

Zatta P, Lucchini R, van Rensburg SJ, Taylor A.The role of metals in neurodegenerative processes: 

aluminum, manganese, and zinc. Brain Res Bull. 2003 Nov 15;62(1):15-28. 

Zawia NH, Lahiri DK, Cardozo-Pelaez F. Epigenetics, oxidative stress, and Alzheimer disease. Free Radic Biol 

Med. 2009 May 1;46(9):1241-9. Epub 2009 Feb 23.

Nome Stefano Mattioli 

Contatti 

Occupational Medicine, 

S.Orsola-Malpighi University Hospital 

via Pelagio Palagi 9, 

I-40138 Bologna, Italy. 

Tel: +39-051 636 2761 

Fax: +39-051 636 2609 

E-mail: s.mattioli@unibo.it 

Istituto/Dipartimento Section of Occupational Medicine, Department of Internal 

Medicine, Geriatrics and Nephrology, University of Bologna, 

Italy 






Using prevention to reduce the burden of Alzheimer’s 

disease (and other ageing diseases) 

Occupational exposure to Valsalva manoeuvre and risk of Alzheimer 

disease 

None



2-3 Years (6 months for Study design and questionnaire preparation; 

at least 1 year for data collection; 6 month for data cleaning, quality 

controls and statistical analysis; 6 month for writing the manuscript 

and internal revision process) 

Introduction 

Chronic increased intracranial pressure or exposure to repetitive 

intermittent intracranial pressure elevations might contribute to 

Alzheimer’s disease (AD) via damage to the choroid plexus resulting 

from cumulative effects of repetitive intermittent intracranial 

pressure elevations, and leading to a reduction in cerebrospinal 

fluid (CSF) production, and hence to diminished CSF clearance of 

neurotoxins such as β-amyloid [Wostyn, Can chronic increased 

intracranial pressure or exposure to repetitive intermittent 

intracranial pressure elevations raise your risk for Alzheimer’s 

disease? Med Hypotheses 2004;62:925–30, 2004]. 

Weightlifters, glassblowers and wind instrument musicians can 

generate very high intracranial pressures due to repeated Valsalva 

manoeuvres (elevated subglottic pressures), which impair venous 

return from the head [Greenfield et al., Transient changes in 

cerebral vascular resistance during the Valsalva maneuver in man. 

Stroke 1984;15:76–9]. Herein, occupational exposure to Valsalva 

manoeuvres might be a risk factor for Alzheimer disease. 

Our aim will be the conduction of an etiological study to test the 

hypothesis of causal link between occupational history of frequent 

Valsalva manoeuvres and the onset of Alzheimer diseases in later 

ages. 

Methods 

We think that a case-control study design would be feasible to our 

purpose. Firstly, our hypothesis implies a long latency between 

exposure and disease onset. Moreover, regular lifting is a common 

task among manual workers. In a previous study conducted in 

Bologna, we estimated a lifetime prevalence of manual lifting works 

around 32% [Mattioli et al., Physical exertion (lifting) and retinal 

detachment among people with myopia. Epidemiology. 2008 

Nov;19(6):868-71]. 

Considering the frequency of Alzheimer disease (10,000 new cases 

registered in Emilia-Romagna every year) we can recruit only 

incident cases, in order to minimize recall bias. 

Selection of cases will be conducted in cooperation with 

Neurological Clinic of Bologna; collaboration of relatives will be

asked in order to properly collect information. 

Selection of controls (2 for each case) will be based on random 

sampling from the national health service registries. 

For data collection, we will use a standardized questionnaire 

designed for assessment of a series of potential occupational and 

non-occupational risk factors, based on those proposed in the 

literature. In particular, weight lifting will be assessed both by 

analysis of occupational history and by asking specific questions. 

Multivariate logistic regression analyses will be conducted to study 

the relationship between past occupational exposure to Valsalva 

manoeuvres and Alzheimer disease. 

Conclusions 

Chronic repetitive Valsalva manoeuvres have been proposed has a 

possible cause of Alzheimer disease. Weight lifting, a very common 

task for manual workers, might play an important role in Alzheimer 

diseases aetiology. If confirmed, this hypothesis could have 

important impact on the global burden of the disease. Indeed these 

findings could have significant implications in prevention, with 

important consequences on health and socio-economic fields. 

Epidemiological studies are needed to test this hypothesis and to 

asses other possible etiologic factors that could be implicated in the 

pathologic mechanism [Wostyn, 2004].

Nome Giuseppe Mele (Professore Aggregato di Chimica) 

Contatti 

giuseppe.mele@unisalento.it 

Studio: 0832-297281 

Cellulare: 333-9593228 

Istituto/Dipartimento Dipartimento di Ingegneria dell’Innovazione-Università del Salento 




Titolo progetto (1) 


Durata progetto (1) 

Abstract del progetto (1) 

Costruzione di un “Alzheimer Web-GIS” che consenta di georeferenziare 

casi di morbo di Alzheimer su aree campione 

associando ad ogni caso in esame una “base di dati” contenente 

parametri geografici e ambientali allo scopo di capire “se possibile” le 

correlazioni tra i parametri (geografici, scientifici, medici, sociali) ed i 

casi oggetto di studio. 

Nella costruzione del “Web_GIS”, sebbene in ambiti diversi, si potrebbe 

trasferire il know-how acquisito nei due progetti di seguito riportati di cui 

il proponente è stato Responsabile Scientifico. 

Acronimo ECODO-NET 

"Development of a model Web based virtual observatory of Acherontas, 

Kalamas and Torre Guaceto ecosystems and its application as a mobile 

exhibit and permanent environmental kiosk towards public awareness 

and sustainable development of coastal ecosystems (EcoDo-net)" 

focuses on the improvement of the management, the protection and the 

sustainable development of the Greek and Italian coastal ecosystems in 

the border area (Acherontas, Kalamas, their intermediary coastal zone 

and -Torre Guaceto) through the observation of the ecosystems' quality 

and the increase of public awareness about environmental issues. 

Programma Interreg IIIA Italia-Grecia 2000-2006 

Asse III. Ambiente e Patrimonio Culturale 

Misura 3.1 Miglioramento della gestione degli ecosistemi 

FESR-Stato-Regione Puglia 

2 anni gennaio /2007-Dicembre/2008 

The program "Development of a model Web based virtual observatory of 

Acherontas, Kalamas and Torre Guaceto ecosystems and its application as 

a mobile exhibit and permanent environmental kiosk towards public 

awareness and sustainable development of coastal ecosystems (EcoDonet)" 

focuses on the improvement of the management, the protection 

and the sustainable development of the Greek and Italian coastal 

ecosystems in the border area (Acherontas, Kalamas, their intermediary 

coastal zone and -Torre Guaceto) through the observation of the 

ecosystems' quality and the increase of public awareness about 

environmental issues.

Titolo progetto (2) 


Durata progetto(2) 

Abstract del progetto (2) 

Linee di intervento per la valorizzazione delle colture agricole locali della 

Valle della Cupa 

-Web GIS (Geographic Information System) as support for sustainable 

development local economy in the fields of agro-industry and fine 

chemicals 

Consorzio Universitario Interprovinciale Salentino 

Consorzio di Comuni della Valle della Cupa 

1 anno 2006 

There is nowadays an increasing interest in the development of natural 

products and new fine chemicals based on renewable organic materials 

using sustainable processes. At the same time innovative developments 

associated to main local productions (e.g. olive oil and wine industries) as 

well as to the extraction fine chemicals from less important local cultivar 

(e.g. mulberry, black-berry, bilberry etc.), are of topical importance 

especially in view of a sustainable “green chemistry”. 

Emerging technologies for simulating, controlling, and optimizing 

complex systems will be considered, crossing a long list of disciplines such 

as, botanical, chemical, computational, material science and process 

simulation. 

In this context we report our studies concerning a planned sampling of 

local agri-food and non-food species (traditional and/or minor cultivar), 

which could be interesting for this purpose. With this sampling we are 

able to trace the real distribution of local cultivations and to map these 

on a Geographic Information System (G.I.S.). Then, each sample will be 

listed depending on its own biological properties and physical condition 

of the selected cultivar.

Nome Prof. Calogero Caruso MED04 

Contatti 

Immunopatology (preclinical): Calogero Caruso, email: 

immunopatologia@unipa.it 

Neurology: Roberto Monastero, email: roberto.monastero@ki.se 

Oral Medicine: Giuseppina Campisi, email: campisi@odonto.unipa.it 

Istituto/Dipartimento Department of Pathobiology and Medical and Forensic Biotechnologies 



Experimental (preclinical) research 

The preclinical group (Immunopathology Unit) is evaluating the 

role of inflammaging in the expression/progression of the disease. So far 

the alterations of the inflammatory response and the pro-inflammatory 

profile of aged people have been studied as risk factors for AD 

development. This aspect has to be further characterized in terms of the 

comparison of the genes and proteins involved in determining the proinflammatory 

profile in healthy old and very old people and in demented 

(both Alzheimer and non Alzheimer). On the other hand, persistent 

peripheral multibacteria infection such as periodontitis (PD), associated 

with gram-anaerobic bacteria capable of exhibiting localized and systemic 

infections in the host, has been proposed as possible aggravating cofactor 

in subjects with vascular disease. In PD the balance between bacteria and 

host response is altered, resulting in a production of high levels of proinflammatory 

mediators and low levels of anti-inflammatory ones. Data 

regarding the PD association with neurodegenerative disorders are 

lacking; however, it seems plausible that, during PD, cytokines increase in 

AD brain due to stimulation of trigeminal nerve fibres, or directly from 

bacterial products. So, the Immunopathology group in association with 

the Oral Medicine group is going to gain insight into this relationship, 

using biological samples collected from the Neurological group. 

Many studies have reported different alterations of the immune 

system in AD, and the involvement of the acquired branches of the 

immune system. To investigate the systemic signs of immune responses 

in AD, the Immunopathology group is also analyzing the expression of 

activation markers on peripheral blood mononuclear cells from AD 

patients and age-matched healthy controls activated in vitro by 

recombinant amyloid peptide (rAβ42) and their cytokine production. This 

kind of study is also useful to obtain biomarkers of AD for monitoring the 

effectiveness of therapeutic interventions. 

Clinical research 

Risk factors for Mild Cognitive Impairment (MCI) - which may precede 

the clinical detection of AD and dementia - and predictors for the 

conversion from MCI to dementia are currently evaluated by the 

Neurological group. For this purpose, hospital-based data and crosssectional 

as well as prospective data from a population-based cohort are

used. In particular, modifiable (non-genetic) and non-modifiable (genetic) 

risk factors are under evaluation. 

• Concerning modifiable risk factors, three etiologically-driven 

hypotheses are currently carried out: 

1. The role of modifiable vascular risk factors (eg, diabetes, 

hypertension, dyslipidemia, obesity), alone or clustered, in the 

concept of metabolic syndrome are under investigation; 

2. The role of behavioural symptoms, particularly depression, in 

determining the risk of MCI and its progression to dementia/AD 

are under evaluation; 

3. The effects of oxidative stress and vitamin E on the development 

of MCI and dementia/AD, and their effect in the progression from 

MCI to dementia/AD are in assessment. 

• Concerning non-modifiable risk factors, we are going to evaluate 

the role of recently proposed and novel putative genes involved 

in AD. We hypothesize to include a large sample sample of AD 

and control subjects (case:control of 1,000:1,000); accordingly, 

this study would benefit from a valid statistical power in 

detecting novel potential genetic markers for AD. These genes 

will be evaluated alone, in clusters and in conjunction with the 

APOE4 allele. This study will be performed in association with 

Immunopathology Unit. Drugs currently in use to treat AD are 

effective only in 20% of patients; their therapeutic effect is 

predominantly under genetic control. Thus, these data would 

encourage new research in pharmacogenomics. 

Finanziamenti ricevuti Principal Investigator: Dr. Roberto Monastero ; Amount of fund: 590,000 euros 





Epidemiology and risk factors for Mild Cognitive Impairment, Alzheimer's 

disease and dementia. The Zabut Aging Project: 5-year follow-up study on 

a rural Sicilian population 

Ministero della Salute, Italia 

3 years 

We would extend recent research on risk factors for Mild Cognitive 

Impairment (MCI) and Alzheimer's disease (AD) and the progression of 

MCI to dementia in the elderly with a special focus regarding modifiable 

(non-genetic) and non-modifiable (genetic) risk factors. Baseline (n=1930 

subjects) and 5-year population-based data (we are expecting to enrol 

approximately 1500 subjects) will, therefore, be used. Firstly, the role of 

vascular, modifiable vascular risk factors, alone or clustered in the 

concept of metabolic syndrome would be investigated. Secondly, the role 

of behavioural symptoms, particularly depression, in determining the risk 

of MCI and its progression to dementia/AD would be evaluated. Thirdly, 

the effects of oxidative stress and vitamin E on the development of MCI 

and dementia/AD, and their effect in the progression from MCI to 

dementia/AD would be assessed. Lastly, this project would evaluate the 

role of recently proposed and novel putative genes involved in AD.

Roberto Monastero, MD, PhD 

Lab of Epidemiology and Psychology of Aging and Dementia 

Dept of Clinical Neuroscience 

University of Palermo 

Via La Loggia 1 

90129 - Palermo, Italy 

Tel. +39-091-6555185 

Fax. +39-091-6555113 

email. roberto.monastero@ki.se 

roberto.monastero@unipa.it 

Type: Progetto Giovani Ricercatori Ministero Salute, 2007 

Title: Epidemiology and risk factors for Mild Cognitive Impairment, Alzheimer's disease and dementia. The 

Zabut Aging Project: 5-year follow-up study on a rural Sicilian population 

Principal Investigator: Dr. Roberto Monastero 

Amount of fund: 590,000 euros 

Lenght: 3-year project 

Abstract 

We would extend recent research on risk factors for Mild Cognitive Impairment (MCI) and Alzheimer's 

disease (AD) and the progression of MCI to dementia in the elderly with a special focus regarding 

modifiable (non-genetic) and non-modifiable (genetic) risk factors. Baseline (n=1930 subjects) and 5-year 

population-based data (we are expecting to enrol approximately 1500 subjects) will, therefore, be used. 

Firstly, the role of modifiable vascular risk factors, alone or clustered, in the concept of metabolic syndrome 

would be investigated. Secondly, the role of behavioural symptoms, particularly depression, in determining 

the risk of MCI and its progression to dementia/AD would be evaluated. Thirdly, the effects of oxidative 

stress and vitamin E on the development of MCI and dementia/AD, and their effect in the progression from 

MCI to dementia/AD would be assessed. Lastly, this project would evaluate the role of recently proposed 

and novel putative genes involved in AD. Due to the large, hypothesized sample of AD and control subjects 

(about 1,000 for each group), this study would benefit from a valid statistical power in detecting novel 

potential genetic markers for AD. These genes would be evaluated alone, in clusters and in conjunction 

with the APOE4 allele.

Nome Emilio Di Maria, MD PhD 

Contatti 

emilio.dimaria@unige.it; +39 0105634368 

Istituto/Dipartimento Dept. of Neuroscience, Ophthalmology and Genetics - University of 

Genova 

and Division of Medical Genetics, Galliera Hospital, Genova, Italy 



identificata 


Problem statement: The clinical manifestations of Alzheimer’s disease (AD) 

often include behavioural disturbances, such as apathy and mood disorders, 

as well as psychotic symptoms (delusions and hallucinations). These features, 

which significantly affect the clinical outcome, are globally referred to as 

behavioural and psychological symptoms in dementia (BPSD). BPSD are 

recognised to cause a poorer course of disease, contributing to increase the 

social, pharmacological, and managing costs of AD. Family studies have 

reinforced the hypothesis that the occurrence of psychosis in AD patients is 

partially determined by genetic factors. 

The present proposal is in accordance to the priorities listed in the letter of 

intent for the joint initiative, namely: 

- Genetic susceptibility to Alzheimer’s disease 

- Biomarkers 

- Studying early onset forms of Alzheimer’s disease to follow-up disease 

progression. 

Objective: to identify genetic risk factors responsible for the occurrence of 

behavioural and neuropsychiatric symptoms in patients with Alzheimer’s 

disease. We intend to pursue this objective by using both main approaches: i) 

genome-wide association studies; ii) candidate gene approach, in that the 

analysis will be performed only on genetic markers which were demonstrated 

to be associated with neurodegeneration or psychiatric disorders. 

To accomplish this plan, a network of Italian clinical centres and laboratory of 

genetics with distinguished expertise in the field has already been built. 

Preliminary results: A large cohort of patients with AD has been recruited 

through a multicentre effort. Patients were assessed at their admittance with 

a complete sociodemographic and clinical data collection. The study protocol 

was approved by the Institutional Review Boards. Patients enrolment is still 

ongoing. By applying the candidate gene approach outlined above, we have 

already demonstrated that the DAOA gene is a genetic risk factor for the 

occurrence of psychotic symptoms in AD [Di Maria et al, J Alzh Dis 2009]. 

Relevance: The identification of genetic factors influencing the occurrence of 

BPSD in AD will shed new light on the disease pathophysiology and allow a 

better prediction of the illness. On the basis of new lines of evidence, earlier 

and tailored treatments could be appropriately administered. 

Note: the Italian network on behavioural and psychological symptoms in dementia, quoted in the proposal 

section, has not been formally assembled. Therefore, the funded project granted to other 

researchers involved could not be reported. 

Titolo progetto MacAge – MCI-AD Conversion: Analysis of Genetic Effects. A cohort




association study on the role of tau gene haplotypes and ApoE in 

determining the evolution from mild cognitive impairment to Alzheimer’s 

disease. 

CARIGE Foundation, Italy 

1 year 

Background: The association of microtubule-associated tau protein gene 

(MAPT) with Alzheimer’s disease (AD) had been controversial, but recent 

literature demonstrated that the MAPT H1c haplotype is a genetic risk 

factor for AD [Myers AJ, 2005]. H1c haplotype is associated with 

increased tau expression and predisposes, in late-onset AD, to the 

unbalance between the 4-repeat- and the 3-repeat-containing isoforms 

(4rep/3rep), thus leading to tau dysfunction and deposition [Myers AJ, 

2007]. A distortion of the 4rep/3rep expression pattern was also found in 

cholinergic neurons from mild cognitive impairment (MCI), a clinical 

condition which is prodromal to AD [Ginsberg SD, 2006]. Reducing tau 

levels in mice protects against amyloid-β-induced neuronal dysfunction 

[Roberson ED, 2007]. 

Preliminary results: We previously demonstrated that the tau H1 

haplotype is a genetic risk factor for corticobasal degeneration, a sporadic 

tauopathy [Di Maria E, 2000]. More recently, a longitudinal study (PI: 

Prof. M. Tabaton) characterised individuals with amnestic MCI (aMCI), in 

order to find biological markers associated with the risk of AD [Assini A, 

2004; Odetti P, 2005]. To date, 224 individuals with aMCI were recruited. 

We foresee to collect within the first phase of the project a cohort of at 

least 240 cases: all followed-up for 1 year, about ⅓ for ≥2 years. 

Objective: To evaluate the hypotheses that the evolution from MCI to AD 

is influenced by the tau locus, and that its effect is mediated by a 

qualitative and quantitative unbalance in the expression of the tau gene 

isoforms. The specific aims will be accomplished according to the 

following experimental plan (total duration: 1 year). 

- Patients evaluation: i) patients follow-up, blood sampling (according 

to the current protocol for the longitudinal study); ii) DNA, plasma, 

serum, cell lines banking (Galliera Genetic Bank). 

- Cohort association study: i) the tau tagging polymorphisms will be 

genotyped in the case series; ii) the association study will investigate 

the effect of the tau haplotypes on the MCI-AD transition, taking into 

account ApoE genotypes, as well as age, age at onset, gender and 

possible interacting variables. 

- Gene expression study: expression level of the tau isoforms will be 

investigated in lymphoblasts from patients belonging to the two 

subsets (MCI and MCI-AD converted) and carrying different tau 

genotypes. 

Relevance and deliverables: The identification of genetic factors 

influencing the evolution from MCI to AD will shed new light on the 

disease pathophysiology and allow a better prediction of the disease 

progression. On the basis of new lines of evidence, earlier and tailored

therapeutic approaches can be envisaged.

Nome Prof. Carlo Caltagirone 

Contatti 

Tel. 

E mail 

Via Aretina, 306 – 00179 Rome 

05 51501409 


Istituto/Dipartimento IRCCS Fondazione Santa Lucia – Laboratory of Clinical and Behavioral 

Neurology. 


Area di interesse identificata Preclinical Diagnosis 






Markers to diagnose neurodegenerative diseases and of new targets to 

test the pharmacological treatments. 


2006-2008 

The present investigation involved 200 individual with AD, whose 

cognitive functioning was monitored along a 18-months period by means 

of administration of the Mini Mental State Examination. The results 

document, on one side, a progressive decline of cognitive performance 

and, on the other side, a significant relationship between the occurrence 

of some specific genome variants and both the severity and the earlier 

onset of disease. In particular, the presence of i) allele e4 of ApoE 

polymorphism, of ii) allele T of the polymorphism of the serotonin 

receptor 5HT2A (102T/C), of iii) some variants of GST genes as well as of 

the iv) polymorphism -137C/C of the promoter region for IL-18, 

significantly influence both the onset and the clinical disease course. 

Moreover, in line with the above data, the analysis of the cellular 

components of innate immunity confirmed that the AD patients’ immune 

system cells are more likely to produce phlogistic factors such as IL-6 and 

IL-18.

Furthermore, it has also been showed that the previously reported 

decrease of proteinic level of ADAM10 within the AD patients’ platelets, is 

not owed to reduced levels of ADAM10 mRNA but, rather, to others 

events that involve the translation processes or the proteolysis of the 

molecule itself. 

Finally, groundbreaking in vivo and in vitro animals studies indicate that 

the Tat-Pro ADAM peptide can produce alterations of the molecular 

composition of the glutamatergic synapses, affecting the synaptic 

functioning. 

In conclusion, the present study adds some valuable information about 

the role played by some disease mediators, such as early inflammatory 

episodes and synaptic changes, thus, helping to identify new molecular 

cascades and cellular processes involved in the pathogenesis of 

neurodegeneration and, likely, to be used as markers of the disease. 

Indeed, by means of project here presented, we were allowed to 

implement an interdisciplinary work to explore the physio-pathogenetic 

pathways in AD, involving a wide number of individuals with AD that 

belong to several Italian centres. As a matter of fact, in the view of the 

multidisciplinary network, relying upon the synergistic coordination 

between basic and clinical researchers, the involvement of the most 

representative ITINAD units allowed us to reach results that significantly 

advance our understanding of AD pathogenesis. Moreover, the same 

results might represent relevant clues to individuate an advanced 

strategy in order both to early diagnose AD and to formulate an accurate 

prognosis.

Nome Manservigi Roberto 

Contatti 

Tel. +39 0532 455401/455412 

Fax +39 0532 974470 

Mail: roberto.manservigi@unife.it 

Istituto/Dipartimento Department of Experimental and Diagnostic Medicine 













Neurodegenerative Diseases 

Search of herpetic proteins involved in the pathogenesis of Alzheimer. 

MiUR 

2006-2008 (two years) 

The overall objective of this proposal is to study the correlation between 

HSV-1 infection and Alzheimer’s disease (AD) and to understand those 

HSV-1 functions that are involved in the pathogenesis of AD. 

: 

Search of herpetic proteins involved in the pathogenesis of Alzheimer's 

disease for the development of new therapeutic strategies 

MiUR 

2008-2010 (two years) 

Despite the very numerous studies on Alzheimer's disease (AD), especially 

on amyloid plaques and neurofibrillary tangles, little information has been 

obtained thus on the cause of the disease. Moreover, different 

experimental evidences define a role for Herpes simplex virus type 1 (HSV- 

1) infection as a risk factor for AD development. The goal of this project is 

to study and characterize the different interactions between viral proteins 

and the disease development, in order to obtain a better understanding of 

AD origins and, above all, to develop innovative therapies.


B. Epidemiology genetics

Nome Luisa Benussi, Giuliano Binetti 

Contatti 

0339-030-3501725 

lbenussi@fatebenefratelli.it 

Istituto/Dipartimento IRCCS “Centro S. Giovanni di Dio-Fatebenefratelli” NeuroBioGen Lab- 

Memory Clinic, Brescia Italy 


Alzheimer disease (AD) is a genetically complex and heterogeneous disorder. Less than 10% of AD patients 

are familial with an autosomal dominant inheritance of the disease (FAD). In autosomal dominant AD 

families, rare mutations in the amyloid precursor protein gene, and the presenilin 1 and 2 genes have been 

identified. In sporadic patients the disease is expected to result from the interaction of multiple 

susceptibility genes and unknown environmental factors. So far, the e4 allele of the apolipoprotein E 

(APOE) gene has been the only consistently replicated genetic risk factor for late-onset AD. Through 

genome wide association studies (GWAS), a novel opportunity of identifying dementia risk genes and 

associated disease pathways has emerged with several potential AD risk genes already reported. Further 

validation and replication in other patients cohort combined with functional characterization of these 

genes could lead to a better understanding of the pathophysiology underlying dementia and open new 

therapeutic possibilities for AD. In the last years our group has contributed to: 1) the identification of 

genetic factors that influence the disease risk and the pharmacological response in AD patients (Nicosia et 

al., Dement Geriatr Cogn Disord. 2001;Ventriglia et al, Mol Psychiatry 2002; Scassellati et al, Neurosci Lett 

2004; Ventriglia et al, Aging Clin Exp Res 2005; Emanuele et al, NBA 2009; Di Maria et al, JAD 2010; Santoro 

et al, CNS Drugs 2010); 2) the identification of mutations that are associated with familial forms of AD 

(Finckh et al, Am J Hum Genet 2000; Finckh et al, Neurology 2000; Binetti et al, Neurosci Lett 2003;Binetti 

et al, Ann Neurol 2003; Signorini et al, Curr Alzheimer Res 2004; Alberici et al, Eur J Neurol 2007;Ghidoni et 

al, JAD 2009); 3) the assessment of areas of intervention for genetic counseling of dementia by a crosscultural 

comparison between Italians and Americans (Binetti et al., PEC 2006). 

The specific objectives of this proposal are: 

1) Identification of genetic factors that influence the disease susceptibility and the pharmacological 

response in AD patients (genome wide association studies); 

2) Identification of new mutations and novel AD-associated loci (genetic screening; linkage analysis; 

quantitative trait - proteomic profiles- linkage analysis); 

3) Evaluating the psychosocial impact of genetic testing in families with hereditary forms of AD. 

To carry out this project our group will collaborate with: Dr Roberta Ghidoni (Proteomics Unit, IRCCS - 

Fatebenefratelli, Brescia) on genetic and proteomic studies; Dr F Tagliavini (IRCCS Fondazione Istituto Carlo 

Besta, Milano), Dr D Galimberti and Dr E Scarpini (University of Milan) on genetics and genetic counseling; 

Prof C Franceschi (Università di Bologna), Dr G Forloni (Istituto Mario Negri, Milano) on genetics and 

pharmacogenetics. 

Our Group, along with the mentioned National collaborators, can offer to the proposed project the 

following resources: 

1) Clinical expertise in Alzheimer disease; 2) A large collection of biological samples from sporadic and 

familial AD patients (more than 1000 index patients) and a similar number of age-matched controls; 3) A 

large collection of families with autosomal dominant inheritance trait for AD (more than 90 families); 4) 

Genetic testing as well as a genetic counselling program for families with hereditary forms of dementia. 

Area di interesse identificata Genetic susceptibility to Alzheimer’s disease and genome wide 




Project1: Validation of genetic and biochemical markers for early 

diagnosis of AD and the prediction of conversion of MCI into AD, and




design of a multivariate molecular protocol having high diagnostic and 

prognostic accuracy. 

Project2: Marker biologici di neurodegenerazione utilizzabili nella pratica 

clinica ai fini della diagnosi precoce e differenziale, della individuazione 

delle forme con risposta ottimale alle attuali terapie 

Project3:GenEtica- Profili bioetici e biogiuridici della genetica tra ricerca 

sperimentale, consulenza e prospettive terapeutiche. 

Project1:Italian Ministry of Health Progr Strategico PS39, prog 1 

Project2:Italian Ministry of Health Progr Strategico conv.71 prog 6 

Project3: Italian Ministry of University and Research, FIRB2006 

Project1:01-01-2009/31-12-2010 

Project2:7-01-2008/7-04-2010 

Project3:15-10-2008/15/10/2011 

Project 1 and Project 2 aims: 

1) The development and validation of a multi-factorial protocol that 

integrates molecular, imaging, neurophysiological, neuropsychological 

and behavioral data for early diagnosis of AD, in particular during the 

preclinical phase and the prodromal stage of "mild cognitive impairment" 

when the symptoms are not severe enough to fulfill the current 

diagnostic criteria for AD. 

2) Validation of this new diagnostic protocol within a regional public 

health network, and evaluation of the health care, organization and 

economic implications of its transfer to the National Health Service. 

Project3 aims: 

1) to evaluate the psychosocial impact of genetic testing in families with 

hereditary forms of dementia; 2) to evaluate the efficacy of the pre-test 

educational session and the information comprehension during genetic 

counselling 

3) analysis of the ethical issues arising from a genetic counselling service 

for dementia

Nome 

Maria Del Zompo 

Contatti 

Phone: +39 070 6092438 

Fax: +39 070 653584 

Email: delzompo@unica.it 

Istituto/Dipartimento Department of Neurosciences “B.B. Brodie”, University of Cagliari, 

Cagliari, Italy 



- Genetic susceptibility to Alzheimer’s disease and genome 

wide association studies 


(In the Biobank of the Section of Clinical Pharmacolgy of the Department 

of Neuroscience, University of Cagliary, are cryopreserved the 

lymphoblastoid cell lines of 147 patients with Alzheimer's disease and 116 

controls. AD patients of proven Sardinian ancestry (parents, grandparents 

and great-grandparents) were diagnosed according to DSM-IV, and 

National Institute of Neurologic and Communicative Disorders and 

Stroke–AD and Related Disorders Association (NINCDS-ADRDA) criteria 

for possible or probable AD (McKhann et al, 1984 ). Cognitive screening 

was performed by means of Mini-Mental State Examination (MMSE) 

(Folstein et al, 1975). All control subjects were cognitively intact 

volunteers with an MMSE score ≥ 26 and with negative family history for 

dementia.)


C. Biomarkers and early diagnosis and imaging

Nome Patrizia Mecocci 

Contatti 

mecocci@unipg.it 

+39 075 5783270 

Istituto/Dipartimento Sezione di Gerontologia e Geriatria 

Dipartimento di Medicina Clinica e Sperimentale 

Università degli Studi di Perugia 


Area di interesse 1)Studies on plasma biomarkers of oxidative/nitrosative stress, inflammation, 

identificata 

lipid and glucose metabolism, in older adults, both in population based and 

clinical based longitudinal studies. Samples (already collected or to be 

collected) derived from Italy and other European countries, with which we are 

collaborating, to develop easily available biomarkers that can be used in the 

elderly to: a) study age-related changes in metabolism, also in relation to 

lifestyle (i.e. diet, physical activity); b) obtain diagnostic and prognostic tools, 

which can be used in different age-related disorders (cognitive 

decline/dementia; multimorbidity; disability). The main goal is to obtain noninvasive 

tools that can be used in age-related disorders for i) investigating 

pathogenetic mechanisms; ii) diagnosis, prognosis and monitoring of 

preventative and therapeutic interventions. 

Age-related cognitive decline and dementia are one of our main research areas, 

and our studies integrated plasma biomarkers and neuroimaging data. 

2) Studie on cellular and animal models of aging, using markers of 

oxidative/nitrosative stress and mitochondrial structure and activity to evaluate 

the effects of novel therapeutics (chemically modified antioxidants, vitamins, 

nutriceuticals) that can be used in humans to prevent age-related cognitive 

disorders. 


Titolo progetto ZINCAGE Contract n° 506850 

Ente finanziatore European Union in the 6th Framework Program (FP6): "Food Quality and 

Safety", Priority 5, Thematic Area T6 


Abstract del progetto This is an epidemiological study on the influence of diet and lifestyle on healthy 

ageing, aimed at preventing adult degenerative disease, particularly focusing on 

cardiovascular and neurodegenerative diseases and also addressing 

malnutrition of the elderly. 


Titolo progetto InnoMed (Innovative Medicines for Europe) Project. Contract n° 518170. 

Ente finanziatore European Union in the 6th Framework Program (FP6): Life Science Health, 

Priority 5. 


Abstract del progetto The project, led by the European Federation of Pharmaceutical Industry and 

Associations (EFPIA), addresses the complex issues associated with the future of 

biomedical research in the EU,and addresses ways of achieving accelerated 

development of, safe and more effective medicines, aiming to revitalize the 

European biopharmaceutical research environment. The project is mainly 

devoted to study cognitive impairment in the elderly and Alzheimer’s disease. 


Titolo progetto K4Care Project (Knowledge-Based Homecare eServices for an Ageing Europe)

Contract n° 026968 

Ente finanziatore European Union in the 6th Framework Program (FP6): Information, Society, 

Technology, Priority 2 


Abstract del progetto The main objective of the K4CARE project is to improve the capabilities of the 

new EU society to manage and respond to the increasing number of senior 

population requiring a personalized healthcare assistance. The project will 

capture and integrate the information, skills, expertises, and experiences of 

specialised centres and professionals of several old and new EU countries, and 

will incorporate them in an intelligent web platform in order to give service to 

health professionals, patients, and citizens in general. Chronic conditions, such 

as dementia, are the main focus of the project. 


Titolo progetto PRIN Oxidative and nitrosative damage of mitochondrial proteins in models of 

cell senescence and in subjects with Alzheimer's disease: a pre-clinical study on 

natural forms and new synthetic analogues of vitamin E 



Abstract del progetto Mitochondria are key targets of oxidative stress with relevance in cell 

senescence (mitochondrial theory of aging) and possibly in the pathogenetic 

mechanisms of age-related diseases such as Mild Cognitive Impairment 

(MCI)and Alzheimer's disease (AD). In this context emerging aspects deal with 

the study of post-transductional modifications (PTMs) in proteins caused by 

reactive oxygen or nitrogen species (ROS and RNS, respectively). The aim of this 

project is to evaluate PTMs in mitochondrial proteins in "in vitro" models of 

cellular senescence as well as in peripheral cells from subjects with MCI and AD 

also evaluating the role as protecting agents of natural forms and newly 

synthesized analogues of vitamin E. 



Young researchers 2007 Epidemiology and risk factors for Mild Cognitive 

Impairment, Alzheimer’s disease and dementia. The Zabut Aging Project: 

5-year follow-up study on a rural Sicilian population. 





3 years 

The study will evaluate the risk factors for MCI and AD and the 

progression of MCI to dementia in the elderly with a special focus 

regarding modifiable (non-genetic) and non-modifiable (genetic) risk 

factors. Baseline and 5-year population-based data will, therefore, be 

used. Main topics are i) the role of modifiable vascular risk factors; ii) the 

role of behavioural symptoms, particularly depression, in determining the 

risk of MCI and its progression to dementia/AD; iii) the effects of 

oxidative stress and vitamin E on the development of MCI and 

dementia/AD; iv) the role of recently proposed and novel putative genes 

involved in would encourage new research in pharmacogenomics.

Nome Prof. Salvatore Monaco 

Contatti 

salvatore.monaco@univr.it 

045-8124285/4461 

Istituto/Dipartimento Neuropatologia/Scienze Neurologiche e della Visione 


Dementia is a clinical syndrome characterised by a progressive loss of cognitive abilities and executive 

functions, with a significant compromise of patient’s social, relational and working functions. Recent data 

suggest that pathological and biochemical deteriorations take place years before the clinical emergence of 

the different dementia conditions. The search for novel markers with a high diagnostic sensitivity is 

therefore one of the major and high-priority objective of dementia research. There are several 

methodologies that are presently used or under development, including proteomics which has been 

shown as one of the most promising and useful for the identification of novel markers. 

Aim of the present program is therefore to identify clinical and biological markers of dementia according 

to the following experimental strategy: 

A). To study mild cognitive impairment and Alzheimer’s disease patients by assessing a specific clinical, 

neuropsychological, genetic and biochemical profile at the individual level. 

B). To identify novel genotypic markers associated to the specific clinical profile from all individual 

patients. 

C). To identify novel CSF and plasmatic phenotypic markers from all patients, by using a proteomic 

approach. 







Biomarkers 

Biobanking 

Translational research 

Disabilità cognitiva e comportamentale nelle demenze e nelle psicosi 

Fondazione CARIVERONA 

3 anni

Nome Rita Casadio, Pier Luigi Martelli 

Contatti 

casadio@biocomp.unibo.it 

gigi@biocomp.unibo.it 

Istituto/Dipartimento Dipartimento di Biologia Evoluzionistica Sperimentale 


Genetic association studies have contributed lists of genetic variations (mainly SNPs) that are 

putatively related with the insurgence and/or the prognosis of the Alzheimer disease (AD). Next 

generation sequencing techniques will enormously increase the amount of data on genome wide 

variability associated with the AD. These data will be useful for discovering new genetic markers of 

the risk and for better understanding the molecular mechanisms underlying the aetiology of the AD. 

To benefit from these data, an important analysis effort has to be done in two different and 

complementary directions: 

1) prioritisation, for highlighting among all the reported variations those that are more likely to 

be related to the AD; 

2) annotation, for understanding if the variations occur in coding or in regulatory regions of the 

genome and for discovering their likely molecular effects. 

Computational methods that are able to integrate information from different sources in a Systems 

Biology perspective are then essential. In particular we propose to implement a pipeline able to: 

[DA 1] search if the variations occur in a protein coding region, in a RNA-coding region or in a 

regulatory element; 

[DA 2] predict the effect of protein mutations on localization, structure, stability, and 

function; 

[DA 3] predict the correlation between protein mutations and diseases; 

[DA 4] predict the effect of protein mutations on the protein-protein and protein-DNA 

interactions; 

[DA 5] identify the mRNAs targeted by miRNAs and to predict the effect of variations in 

miRNAs; 

[DA 6] predict the effect of variations in regulatory regions on the binding with transcription 

factors; 

[DA 7] analyse the effect of cariations on regulatory and interaction networks and on 

metabolic and signalling pathways; 

[DA 8] collect, compare and store the results of previous analyses for different variations. 

The Biocomputing Group of the University of Bologna developed several methods that can be

integrated in the pipeline (see www.biocomp.unibo.it). Among the others, efficient tools of interest in 

this particular case are devoted to: 

1) the structural and functional annotation of protein sequences (BAR: Bartoli et al. 2009); 

2) the prediction of membrane protein topology (ENSEMBLE: Martelli et al. 2003); 

3) the prediction of perturbations in protein stability upon mutations (I-Mutant: Capriotti et al. 

2008); 

4) the prediction of the correlation between protein mutations and diseases (SNPs&GO: 

Calabrese et al. 2009); 

5) the protein localization (BaCelLo: Pierleoni et al. 2009) 

6) the prediction of protein interaction sites (ISpred: Fariselli et al. 2002) and their analysis in the 

context of genome-wide interaction networks (Bartoli et al., in press) 

7) the prediction of silencing effect of miRNAs (Montanucci et al. 2008). 

References 

� Bartoli L, Montanucci L, Fronza R, Martelli PL, Fariselli P, Carota L, Donvito G, Maggi G, 

Casadio R -The Bologna Annotation Resource: a non-hierarchical method for the functional 

and structural annotation of protein sequences relying on a comparative large-scale genome 

analysis- J Proteome Res 8:4362-4371 (2009) 

� Bartoli L, Martelli PL, Rossi I, Fariselli P, Casadio R -The prediction of protein-protein 

interaction sites in genome-wide protein interaction networks: the test case of the human cell 

cycle- Curr Prot Pept Sci (in press) 

� Calabrese R, Capriotti E, Fariselli P, Martelli PL, Casadio R -Functional annotations improve the 

predictive score of human disease-related mutations in proteins- Hum Mutat 30:1237-1244 

(2009) 

� Capriotti E, Fariselli P, Rossi I, Casadio R -A three-state prediction of single point mutations on 

protein stability changes- BMC Bioinformatics 9 Suppl 2:S6 (2008) 

� Fariselli P, Pazos F, Valencia A, Casadio R -Prediction of protein-protein interaction sites in 

heterocomplexes with neural networks- Eur J Biochem 269:1356-1361 (2002) 

� Martelli PL, Fariselli P, Casadio R -An ENSEMBLE machine learning approach for the prediction 

of all-alpha membrane proteins- Bioinformatics 19:I205-I211 (2003) 

� Montanucci L, Fariselli P, Martelli PL, Rossi I, Casadio R -In Silico Evidence of the Relationship 

Between miRNAs and siRNAs- Open Appl Inf Journal 2:9-13 (2008) 

� Pierleoni A, Martelli PL, Fariselli P, Casadio R -BaCelLo: a balanced subcellular localization 

predictor- Bioinformatics 22:e408-e416 (2006) 

Area di interesse identificata Genome wide association studies

Nome 

Elio Scarpini, Daniela Galimberti 

Contatti 

0255033847 

daniela.galimberti@unimi.it 

Istituto/Dipartimento Università di Milano, Dipartimento di Scienze Neurologiche 


Given the rationale that AD is a multifactorial disease and that inflammation plays a role in its 

pathogenesis, this proposal aims to study in detail genes encoding for inflammatory molecules, including 

progranulin and chemokines, all of which located in chromosome 17q.21, both from a genetic and 

functional point of view. This research will take advantage from the availability of a biobank including 500 

patients with AD diagnosed according to current criteria, and a similar number of age-matched controls. 

Genomic DNA have been collected for each subject. In addition, in a group of about 200 patients and 

controls, serum, plasma, CSF and RNA from PBMC and CSF cells have been collected as well. 

In particular, the specific objectives of this research are aimed to: 

1. Carry out a thorough association analysis of the region of chromosome 17 containing PGRN and the 

selected chemokine cluster (CCL2, CCL3, CCL4, CCL5, CCL7, CCL8, CCL11, CCL13, CCL14, CCL15, 

CCL16, CCL18, CCL23, CCR7, CCR10). Optimal tagging SNPs covering all the linkage blocks will be 

chosen. To this aim, we will perform a power calculation using the Genetic Power Calculator 

program (http://pngu.mgh.harvard.edu/~purcell/gpc/) based on sample size, the average observed 

MAF, and assuming a multiplicative model and an estimated prevalence of AD in Italy of 1 case per 

1000 inhabitants after 65 years of age. 

2. Evaluate expression levels of the above mentioned genes in cells isolated from CSF and PBMC by 

Real-Time PCR. Prioritized hits selected at aim 1) will be correlated with expression levels to test 

whether these SNPs influence the level of transcription. 

3. Analyze the pattern of miRNA which act as silencer of mRNA studied in aim 2). miRNA possibly 

influencing the translation of the above mentioned genes will be selected by submitting queries 

at the website http://microrna.sanger.ac.uk/, which contains sequences of all published mature 

miRNA, together with their predicted source hairpin precursors. 

4. Evalutate, by ELISA, CSF and serum levels of PGRN and chemokines. 

5. Combine genetic, expression and functional data with the clinical phenotype of patients, including 

longitudinal full neuropsychological testing, treatment efficacy and brain/hippocampal atrophy at 

MRI, in order to identify a genetic pattern and/or novel pathogenetic mechanisms suitable to 

predict which patients will likely decline more rapidly or will respond to therapy. 

Centers collaborating to this project: University of Brescia (A. Padovani, D. Finazzi), IRCCS 

Fatebenefratelli Brescia (G. Binetti, R. Ghidoni), IRCCS Castellanza (M. Franceschi), IRCCS S. Raffaele, 

Milan (S. Cappa), University of Turin (I. Rainero). From the University of Milan: C. Mariani, R. Dominici, 




Genetic susceptibility to Alzheimer’s disease and genome wide 


- Genetic Risk factors and peripheral biological markers of conversion 

from Mild Cognitive Impairment to Alzheimer's Disease (to DG) 

- Introducing new bio-markers in clinical practice for the early diagnosis of 

Alzheimer disease: methodological, clinical and organisational aspects for




the National Health System (to ES) 

Progetti ( 1 and 4) parte di Programma Strategico (PS39), Italian Ministry 

of Health 

01-01-2009/31-12-2010 

This program has two primary aims: 

A. The development and validation of a multi-factorial protocol that 

integrates molecular, imaging, neurophysiological, neuropsychological 

and behavioral data for early diagnosis of Alzheimer's disease (AD), in 

particular during the preclinical pha se and the prodromal stage of "mild 

cognitive impairment" (MCI) when the symptoms are not severe enough 

to fulfill the current diagnostic criteria for AD. The achievement of this 

objective is one of the priorities of the international research com munity, 

in the perspective of specific, disease-modifying therapies that are under 

development. 

B. Validation of this new diagnostic protocol within a regional public 

health network, and evaluation of the health care, orga nization and 

economic implications of its transfer to the National Health Service (NHS). 

The program will be articulated into four complementary and strictly 

integrated projects with the following specific aims: 

PROJECT 1. Validation of genetic and biochemical markers for early 

diagnosis of AD and the prediction of conversion of MCI into AD, and 


prognostic accuracy. 

PROJECT 2. Development of standard operating procedures (SOPs) and 

quality control (QC) for neuroimaging (MRI, 18F-FDG PET and 18F-DDNP 

PET) and clinical neurophysiological analysis (including 

electroencephalography, magnetoencephalography and transcranial 

magnetic stimulation) for the diagnosis of prodromic or incipient AD. 

PROJECT 3. Definition of cognitive and behavioral indicators that 

differentiate various forms of MCI and enable the diagno-sis of incipient 

AD. 

PROJECT 4. Validation of a comprehensive diagnostic protocol based on 

the results obtained in the above studies, in a series of Alzheimer 

Evaluation Units of Lombardia Region, and estimation of costs and 

benefits of its transfer to the NHS.

Nome Prof. Paolo Maria Rossini 

Contatti 

Email: paolomaria.rossini@afar.it 

Istituto/Dipartimento Neurology, University “Campus Biomedico”, Rome, Italy 


Individual evaluation of cognitive decline: definition of a low-cost non-invasive examination battery and 

implementation of a predictive algorithm to identify Mild cognitive impairment. 


Mild cognitive impairment (MCI) is a state of the elderly brain 

affecting a large number of individuals mainly characterized by memory 

impairment not yet encompassing the definition of dementia and, in the 

majority of cases, represents a precursor of Alzheimers’ Disease (AD). 

Because of MCI’s prodromal qualities do not greatly impair daily 

functioning, MCI can be identified only by specific clinical and 

neuropsychological evaluation. Within such a theoretical frame a reliable 

marker of MCI-to-AD progression is not yet available, even if it would be 

highly desirable both for research and health system purposes. Ideally, 

these markers should be non-invasive, widely available and of low-cost in 

order to screen out large population samples. Along this line a 

combination of neurophysiological (electroencephalography, transcranial 

magnetic stimulation), blood chemistries (including metals, pro- and 

antioxidant compounds) and genetics are a potential candidate. . In this 

way we have collected numerous studies (1-10). The line of research we 

are interested in focuses on the study of healthy subjects and patients 

(suffering from MCI, AD and vascular dementia – VaD-) by means of this 

multidimensional investigation and will try to identify prognostic indexes 

able to predict the risk of conversion from MCI to AD. 

References: 1.Babiloni C, Frisoni GB, Vecchio F, Pievani M, Geroldi 

C, De Carli C, Ferri R, Vernieri F, Lizio R, Rossini PM Global functional 

coupling of resting EEG rhythms is related to white-matter lesions along 

the cholinergic tracts in subjects with amnesic mild cognitive impairment. 

J Alzheimers Dis. 2010 Jan;19(3):859-71. 

2.Moretti DV, Frisoni GB, Fracassi C, Pievani M, Geroldi C, Binetti 

G, Rossini PM, Zanetti O. MCI patients' EEGs show group differences 

between those who progress and those who do not progress to 

AD.Neurobiol Aging. 2009 Dec 16. [Epub ahead of print] 

3. Babiloni C, Frisoni GB, Vecchio F, Pievani M, Geroldi C, De Carli 

C, Ferri R, Vernieri F, Lizio R, Rossini PM. Global Functional Coupling of 

Resting EEG Rhythms is Related to White-Matter Lesions Along the 

Cholinergic Tracts in Subjects with Amnesic Mild Cognitive Impairment. J 

Alzheimers Dis. 2009 Nov 17. [Epub ahead of print] 

4. Moretti DV, Pievani M, Geroldi C, Binetti G, Zanetti O, Cotelli 

M, Rossini PM, Frisoni GB. 

Increasing hippocampal atrophy and cerebrovascular damage is 

differently associated with functional cortical coupling in MCI patients. 

Alzheimer Dis Assoc Disord. 2009 Oct-Dec;23(4):323-32. 

5. Moretti DV, Pievani M, Geroldi C, Binetti G, Zanetti O, Rossini






PM, Frisoni GB. 

EEG markers discriminate among different subgroup of patients with mild 

cognitive impairment. Am J Alzheimers Dis Other Demen. 2010 

Feb;25(1):58-73. Epub 2009 Feb 9. 

6. Moretti DV, Fracassi C, Pievani M, Geroldi C, Binetti G, Zanetti 

O, Sosta K, Rossini PM, Frisoni GB. Increase of theta/gamma ratio is 

associated with memory impairment. Clin Neurophysiol. 2009 

Feb;120(2):295-303. Epub 2009 Jan 1. 

7. Babiloni C, Pievani M, Vecchio F, Geroldi C, Eusebi F, Fracassi C, 

Fletcher E, De Carli C, Boccardi M, Rossini PM, Frisoni GB. White-matter 

lesions along the cholinergic tracts are related to cortical sources of EEG 

rhythms in amnesic mild cognitive impairment. Hum Brain Mapp. 2009 

May;30(5):1431-43. 

8. Babiloni C, Visser PJ, Frisoni G, De Deyn PP, Bresciani L, Jelic V, 

Nagels G, Rodriguez G, Rossini PM, Vecchio F, Colombo D, Verhey F, 

Wahlund LO, Nobili F. Cortical sources of resting EEG rhythms in mild 

cognitive impairment and subjective memory complaint. Neurobiol Aging. 

2008 Nov 20. [Epub ahead of print] 

9 Moretti DV, Pievani M, Fracassi C, Geroldi C, Calabria M, De 

Carli CS, Rossini PM, Frisoni GB. Brain vascular damage of cholinergic 

pathways and EEG markers in mild cognitive impairment. J Alzheimers 

Dis. 2008 Nov;15(3):357-72. 

10. Rossini PM, Buscema M, Capriotti M, Grossi E, Rodriguez G, Del Percio 

C, Babiloni C. Is it possible to automatically distinguish resting EEG data of 

normal elderly vs. mild cognitive impairment subjects with high degree of 

accuracy? Clin Neurophysiol. 2008 Jul;119(7):1534-45. Epub 2008 May 15. 

Is it possible to automatically distinguish resting EEG data of normal 

elderly vs. mild cognitive impairment subjects with high degree of 

accuracy? 

AFaR, Associazione Fatebenefratelli per Ricerca Biomedica e Sanitaria 

2005-2006; 2006-2007; 2007-2008; 2008-2009; 2009-2010 

To explore if Implicit function as squashing time (IFAST) based 

electroencephalographic (EEG) can reliably distinguish of mild cognitive 

impairment (MCI) and Alzheimer's disease (AD) subjects with

Nome ORAZIO ZANETTI 

Contatti 

TEL 030 3501 358 

Email: ozanetti@fatebenefratelli.it 

Istituto/Dipartimento U.O.Alzheimer-Centro per la Memoria 

IRCCS Centro S.Giovanni di Dio-Fatebenefratelli, Brescia 


Nell’agosto 2007 un gruppo di studiosi ha proposto nuovi criteri diagnostici per la diagnosi di malattia di 

Alzheimer (AD) prodromica o preclinica (Dubois et al.:Research criteria for the diagnosis of Alzheimer's 

disease: revising the NINCDS-ADRDA criteria. Lancet Neurol. 2007 Aug;6(8):734-46.) incorporando, oltre ai 

dati anamnestici, clinici e neurospicologici l’utilità di marker neuroradiologici (atrofia ippocampale o 

ipoperfusione temporo-parietale alla PET), e liquorali (riduzione della proteina ABeta 42 ed aumento di 

TAU). Dal luglio 2006 è attivo presso l’IRCCS S.Giovanni di Dio Fatebenefratelli un Ambulatorio 

Traslazionale per la Memoria (ATM) che coinvolge tutte le Unità Operative cliniche e di ricerca dell’Istituto 

al fine di offrire, soprattutto ai pazienti lievi un percorso diagnostico in grado di rispondere 

all’interrogativo ciclopico, con risvolti laceranti, che ci veniva posto: “vorrei sapere se ho o non ho la 

malattia di Alzheimer”. Nell’arco di 12 mesi sono stati valutati 144 pazienti consecutivamente afferiti all’ 

ATM, divisi in quattro gruppi sulla base della diagnosi clinica (utilizzando i noti criteri diagnostici NINCDS- 

ADRDA per la AD): 12 pazienti sono affetti da disturbo soggettivo di memoria (SMC, MMSE 28.0±1.1); 37 

da “mild cognitive impairment” (MCI, MMSE 25.1±3.6), 55 da malattia di Alzheimer (MMSE 21.1±3.5), e 40 

da demenze non Alzheimer (MMSE 21.6±5.5)(Vedi tabella). A tutti i pazienti sono state proposte le 

seguenti indagini: 1) la volumetria ippocampale alla MR; 2) l’analisi visiva della PET con 

fluorodesossiglucosio; 3) la concentrazione di Tau totale e di Abeta1-42 sul liquor cerebrospinale. 

La sensibilità di ogni marker è più elevata (43 to 71%) per la malattia di Alzheimer rispetto all’ MCI (18 to 

31%). La specificità dei marker rispetto a SMC e demenze non Alzheimer è risultata buona, essendo 

superiore al’ 83% ed al 69% rispettivamente. La positività di almeno un marker presenta un’elevata 

sensibilità per la AD ed un’elevata specificità per il SMC. L’utilizzo di un numero maggiore di marker (due o 

tre) contemporaneamente aumenta notevolmente la specificità diagnostica a scapito però della sensibilità. 

In conclusione i marker recentemente proposti per la diagnosi precoce di malattia di Alzheimer possono 

essere di utilità nella pratica clinica. La valutazione longitudinale consentirà di valutare meglio il loro 

“valore aggiunto” rispetto alla tradizionale diagnosi che si fonda prevalentemente sull’indagine clinica 

corroborata da una semplice TAC encefalica. Per un oculato impiego delle risorse e delle nuove tecnologie 

è infatti importante stabilire con precisione quali siano i pazienti (verosimilmente coloro con un disturbo 

cognitivo molto lieve) che maggiormente possono trarre vantaggi da una procedura diagnostica complessa

e costosa. 


Finanziamenti ricevuti – € 20.000 

(Programma strategico 2006) 





Impiego dei makers biologici e neuro radiologici nella diagnosi 

differenziale precoce 

Strumenti e procedure diagnostiche per le demenze utilizzabili nella 

clinica ai fini della diagnosi differenziale precoce 

Ministero della Salute – Programma strategico 2006- 

triennale

Nome Sandro Sorbi 

Contatti 







sorbi@unifi.it 

tel +390554298465 

Dept. of Neurological and Psychiatric Sciences, University of Florence, 

Italy 

Research in the field of aging and dementia is focusing on the 

characterization of the early stages of cognitive decline. It has been 

suggested that many patients with Mild cognitive impairment (MCI) 

represents already not recognized AD patients. The purpose of the 

research is to identify tools and diagnostic procedures that can identify 

early stages of the disease. We will study the various domains in order to: 

a) Diagnose and characterize demented patients for a proper 

classification of the forms of dementia (Alzheimer's disease, fronto 

temporal dementia, FTD, dementia with motor signs such as Lewy body 

disease and others) b) Diagnose and characterize patients with Mild 

Cognitive Impairement c) To study the neuropsychological aspects to 

identified early MCI patients, both multi domain and amnesic MCI. d) 

Analyze the genetic patient profile to identify the genetic risk factors 

involved in the development and progression of AD. 

The study of genetic factors for AD will be performed in patients with 

familial forms of dementia and in patients with MCI. In addition 

neuropsychological and neuroimaging evaluations will be performed for 

each patients. The study will be divided into the following phases: - 

analysis of autosomal dominant genetic mutations: new families affected 

by AD will be identified and characterized clinically and genetically. The 

goal will be to identify new causative mutations. the Initial screening of 

molecular diagnosis will be a detailed genetic screening of the genes 

associated with familial form of the disease, presenilins (PS-1 and PS-2) 

and the amyloid precursor protein (APP) - Search for other possible 

factors of genetic susceptibility will be performed on both familial and 

sporadic AD patients. It will be studied allelic variations of genes 

potentially involved in chromosomal regions of interest. It will also be 

carried out the study of Apolipoprotein E genotype (ApoE), considered 

today the only susceptibility factor associated with the disease. 

“Strumenti e procedure diagnostiche per le demenze utilizzabili nella 

clinica ai fini della diagnosi precoce e differenziale, della individuazione 

delle forme a rapida o lenta progressione e delle forme con risposta 

ottimale alle attuali terapie” 

MINISTERO DELLA SALUTE , RICERCA FINALIZZATA 2006













6/1/2008-6/4/2010 

The main aim of the project is to integrate the results of neurocognitive, 

neuropsychological, neuroimaging and genetic studies as markers for the 

differential diagnosis of early dementia. 

GENETIC STUDY ON ALZHEIMER'S DISEASE AND THE INVOLVEMENT OF 

OTHER NON-GENETIC FACTORS THAT MAY INFLUENCE THE CLINICAL 

EXPRESSION OF THE DISEASE 

Progetto di ricerca scientifica d'Ateneo (ex quota 60%) 

2009-2010 

GENETIC STUDY ON ALZHEIMER'S DISEASE 

"Analisi dei fattori genetici coinvolti nelle malattie neurodegenerative: 

Utilizzo e applicazione di tecnologie di avanguardia per lo studio della 

malattia di Alzheimer, della demenza frontotemporale e della malattia di 

Parkinson" /* 

COMPAGNIA SAN PAOLO 

2008-2010 

GENETIC STUDY ON ALZHEIMER'S DISEASE 

Aim of the study is to determine the clinical and genetic features of 

movement disorders in patient affected by dementia and the clinical 

features of cognitive decline in patients affected by movement disorders. 

All the patients recruited will be assessed at baseline and during the 

follow-up with selected clinical scales and neuropsychological tests. 

Beyond movement disorder and cognitive deficit, attention will be paid to 

behavioural disturbances, mood disorders and impulse control disorders 

in these patients. 

Patients with a familial history of movement disorders or dementia will 

undergo genetic analysis on the genes implicated in autosomal dominant 

forms of Parkinson disease including PARK1, PARK4, SNCA and LRRK2. 

All the patients recruited will be submitted to FP-CIT-SPECT in order to 

verify and to measure the presence of nigrostriatal pathway

C. Nome 






Flavio Nobili, Guido Rodriguez 

degeneration. 

We plan to enrol 50-100 patients affected by those neurodegenerative 

disorders that may be characterized by movement disorders and 

cognitive decline: Parkinson’s disease, Alzheimer’s disease with 

parkinsonism, Frontotemporal dementia, Lewy body disease, vascular 

dementia, corticobasal degeneration and progressive supranuclear palsy. 

“ Epidemiologia e sviluppo di strumenti di integrazione socio-sanitaria 

nella gestione dei pazienti affetti da Malattia di Parkinson e da altre 

malattie degenerative con concomitante disturbo motorio e declino 

cognitivo” 

MINISTERO DELLA SALUTE, RICERCA FINALIZZATA 2007 

12/2009-12/2010 

Main aim of the project is to analyze and develop in detail the natural 

history of Parkinson's disease and Parkinsonism due to cognitive decline 

and behavioural disturbances. Epidemiological study is of fundamental 

importance in developing diagnostic and therapeutic strategies and new 

methods of integrating social and health care. 

In particular we will identify and recruit individuals with Parkinson's 

disease and classical subjects with parkinsonism linked to cognitive 

decline, it will estimate the incidence of dementia in the two types of 

disturbances. Will be performed genetic analysis of genes related to 

Parkinson's disease in people with familiarity with the disease. All 

patients will undergo a battery of standardized tests (Unified Parkinson's 

Disease Rating Scale and Hoehn and Yahr and neuropsychological tests) 

and FP-CIT-SPECT imaging.

Contatti 

Flaviomariano.nobili@hsanmartino.it; guido@unige.it 

Tel: 010 3537568; Fax: 010 5556893; cell: 333 8331613 

Istituto/Dipartimento Neurofisiologia Clinica, Dip. di Neuroscienze, Oftalmologia e Genetica, 

Università di Genova 


A. ‘A multimodality index for the early diagnosis of Alzheimer’s disease’ 















Early diagnosis 

‘Descripa’ (http://www.descripa.eu/) 

EU FP-5 

4 years 

The DESCRIPA study aims to develop screening guidelines and clinical 

criteria for Alzheimer’s disease in non-demented subjects. The clinical 

criteria will be based on a prospective cohort study of non-demented 

subjects from a memory clinic. The screening guidelines will be based on 

a meta-analysis of prospective population-based cohort studies in 

Europe. 

‘ICTUS’ (Neuroepidemiology. 2007;29(1-2):29-38.) 

EU FP-5 

3 years 

Observational study of cognitive and behavioural effects of 

acetylcholinesterase inhibitors in patients with mild to moderate 


‘Descripa’ (http://www.descripa.eu/) 

EU FP-5 

4 years 

Abstract del progetto The DESCRIPA study aims to develop screening guidelines and clinical 

criteria for Alzheimer’s disease in non-demented subjects. The clinical 

criteria will be based on a prospective cohort study of non-demented 

subjects from a memory clinic. The screening guidelines will be based on 

a meta-analysis of prospective population-based cohort studies in 

Europe. 

Titolo progetto ‘ICTUS’ (Neuroepidemiology. 2007;29(1-2):29-38.)




EU FP-5 

3 years 

Observational study of cognitive and behavioural effects of 

acetylcholinesterase inhibitors in patients with mild to moderate 


BACKGROUND. During the last decade, there has been a great effort to improve diagnostic accuracy of 

Alzheimer’s disease (AD) since the earliest stages. The main advances have been observed with 

morphological Magnetic Resonance (MR) and functional (FDG-PET) brain imaging, and with Cereborspinal 

fluid (CSF) assays of Tau protein and β amyloid fragments. The accuracy of these tools is rather good when 

comparing patients with overt AD dementia versus healthy controls, but decreases in the very early stages 

of the disease. In these stages, the subjects still maintain full autonomy in everyday life, but subtle deficit in 

memory and/or other cognitive domains can already be shown by cognitive tests (Mild Cognitive 

Impairment, MCI). Moreover, some patients complain of memory disturbances that cannot be confirmed 

by cognitive assessments (subjective memory complaints, SMC) but that may sometimes be prodromic of 

AD. 

Pharmacological research is currently very active in trying to identify drugs that can modify the 

natural course of the disease (‘disease-modifying’). While they are tested mainly in overt AD dementia at 

present, it is obvious that effective drugs will be used as soon as possible in the course of the disease, i.e., 

at the MCI stage, or even earlier. 

Thus, an early diagnosis is of paramount importance to start effective treatment as soon as these 

drugs will be available. Yet, individual diagnostic accuracy of functional and morphological neuroimaging 

and CSF assays in these early stages is not fully satisfactory versus healthy controls but even more versus 

the other commonest forms of dementia, such as dementia with Lewy bodies (DLB), frontotemporal 

dementia (FTD) and vascular dementia (VaD). The more recent amyloid PET imaging has added value to 

neuroimaging, showing very high sensitivity but limited specificity versus healthy controls, DLB and 

Parkinson’s disease dementia. 

The reasons of incomplete accuracy are complex and partially understood. Genetic heterogeneity, 

environmental factors, and phenotipic variants of AD may explain incomplete sensitivity on the one hand. 

On the other hand, overlapping atrophy, hypometabolic and biochemical patterns with FTD, DLB and VaD 

may account for incomplete specificity of MR, PET, and CSF assays, respectively. 

The idea that the ‘shadow’ area of each diagnostic tool could be 'unmasked' by the other tools has 

moved researchers to join together the data deriving from more than one modality. Some work has already 

been done, showing an increase accuracy when two examinations are employed together. In this context, 

the ‘core’ cognitive deficit of AD, i.e., verbal episodic memory delayed recall, has also been included. 

Previous attempts to obtain combination indexes include neuropsychology and PET (1-2), neuropsychology, 

PET and Apolipoprotein E (ApoE) genotype (3), neuropsychology and MR imaging (4), Single Photon 

Emission Computed Tomography (SPECT) and MR imaging (5), CSF markers and SPECT (6). However, the 

data are still largely incomplete, especially regarding the very early stages of the disease. 

AIM. To test the hypothesis that joining together the information deriving from MR and FDG-PET imaging, 

memory tests, CSF assays and genetic predisposing factors (such as the ApoE genotype) can improve 

diagnostic accuracy in patients with AD dementia at the MCI stage. A single index could be achieved with 

integrated accuracy deriving from each tool. 

METHODS. Patients. A multicentric approach is needed to reach large patient series in a limited time. 

Patients with MCI of the amnestic type (aMCI), as well as age-matched healthy controls, should be 

recruited by Centres with proven expertise in dementia diagnosis and management. These subjects should 

undergo full neuropsychological examination (common minimum battery to be agreed), MR imaging, and

FDG-PET. CSF assays are performed in MCI patients and in that part of healthy controls giving the informed 

consent for this invasive procedure, providing Ethic Committees approve this manoeuvre in healthy 

subjects. Administration of radiopharmaceuticals to healthy subjects must also be allowed by Ethic 

Committees and by the National Authority governing radiation exposure to healthy subjects for research 

purposes. 

Then, both patients and controls are regularly followed by means of clinical and neuropsychological 

examinations on yearly basis for at least three years to pick up those MCI patients matching the current 

criteria for the diagnosis of AD or other dementias. Other study groups are represented by patients 

affected with FTD, VaD or LBD in a mild stage (i.e., MMSE score >20), recruited by the same centres, 

following the internationally accepted guidelines. 

Image analysis. Several software are available to automatically segment the whole brain or specific brain 

regions, such as the hippocampus and the inferior parietal lobule, that better distinguish AD patients and 

controls. These software can either run both MR and PET images (such as Statistical parametric Mapping, 

SPM) or specifically one of the two (such as the boundary shift integral for MR and ‘Neurostat’ for PET). As 

for MR images, the output could be a volumetric index. The same applies to FDG-PET where a metabolic 

index can be automatically computed. For both MR and PET, other indexes expressing the probability to 

have maximum or minimum atrophy/hypometabolism versus a control group can be computed. 

Statistical analysis. Statistics should rely on a multidimensional approach able to identify the best 

combination of indexes that distinguish between AD patients (at MCI stage) and either healthy controls or 

DLB, FTD, and VaD, respectively. Stepwise and discriminant approaches are favourites. 

EXPECTED RESULTS. An index able to discriminate AD from healthy controls and from the other 

commonest foms of dementia with very high accuracy, i.e., >95%. This index should work, even if with a 

more limited accuracy, in those cases where just some of the requested information is available, for 

instance just with neuropsychology, ApoE, MR and PET images, without CSF biomarkers. Once established, 

such an index may be applied also to subjects with SMC to unveil AD pathology, as well as to screen 

population at high risk of AD, such as direct relatives of AD patients. 

REFERENCES. (1) Anchisi D, et al. Heterogeneity of brain glucose metabolism in mild cognitive impairment 

and clinical progression to Alzheimer disease. Arch Neurol 2005;62:1728-33. (2) Nobili F, et al. Principal 

component analysis of FDG PET in amnestic MCI. Eur J Nucl Med Mol Imaging 2008;35:2191-2202. (3) 

Mosconi L, et al. MCI conversion to dementia and the APOE genotype. A prediction study with FDG-PET. 

Neurology 2004;63:2332-40. (4) Visser PJ, et al. Medial temporal lobe atrophy and memory dysfunction as 

predictors for dementia in subjects with mild cognitive impairment. J Neurol 1999;246:477-85. (5) El Fakhri 

G, et al. MRI-guided SPECT perfusion measures and volumetric MRI in prodromal Alzheimer disease. Arch 

Neurol. 2003;60:1066-72. (6) Okamura N et al. Combined Analysis of CSF Tau Levels and 

[(123)I]Iodoamphetamine SPECT in Mild Cognitive Impairment: Implications for a Novel Predictor of 

Alzheimer's Disease. Am J Psychiatry. 2002;159:474-6. (7) Barnes J, et al. Automatic calculation of 

hippocampal atrophy rates using a hippocampal template and the boundary shift integral. Neurobiol Aging. 

2007;28:1657-63. (8) Ishii K, et al. Statistical brain mapping of 18F-FDG PET in Alzheimer's disease: 

validation of anatomic standardization for atrophied brains. J Nucl Med. 2001;42:548-57.

Nome Professor Paolo Caffarra, M.D. 

Contatti 

e-mail: paolo.caffarra@unipr.it 

phone/fax: 0039-0521-704116 

Istituto/Dipartimento Department of Neuroscience 


Via Gramsci 14 

43100 Parma 

Italy 



The need of an early diagnosis is essential for promoting a strategic 

pharmacological and non-pharmacological intervention on Alzheimer’s 

disease (AD). The identification of cognitive and neurobiological 

markers which might represent salient pre-clinical predictors of the 

insurgence of AD is essential nowadays. Our research focuses on the 

discovery of cognitive instruments useful for a differential diagnosis of 

AD, such as memory and executive tasks, and neuroimaging markers 

using different techniques, such as magnetic resonance imaging (MRI) 

and positron emission tomography (PET). Recently, we have set up an 

innovative and computerized experimental procedure to score semantic 

abilities in patients suffering from Mild Cognitive Impairment (MCI) and 

mild AD (Arango-Lasprilla et al., 2007; Ahmed et al., 2008) , investigating 

also the functional and metabolic neural substrates using MRI and F 18 - 

FDG-PET. Different aspects of structural MRI will be observed, such as 

volumetric analysis, diffusion tensor imaging (DTI), imaging of default 

mode at rest and imaging of the deposition of iron in memory-related 

critical brain regions, i.e. hippocampus. In particular, the observation of 

iron deposition in the brain of patients at risk for AD, represents an 

innovative approach in the neuroimaging field of dementia (Ding et al., 

2009). F 18 -FDG-PET constitutes an effective technique in the monitoring 

of early Alzheimer’s disease progression (Fouquet et al., 2009). 

Cognitive and neuroimaging follow-up data will be informative on the 

identification of specific possible markers of conversion (Devanand et 

al., 2007; McGeown et al., 2009). The role of the polymorphism of APOE 

is observed and correlated with clinical progression of the disease, 

cognitive performance and neuroimaging pattern. 

References: 

Ahmed S, Arnold R, Thompson SA, Graham KS, Hodges JR. (2008). 

Naming of objects, faces and buildings in mild cognitive impairment. 

Cortex, 44, 746-52. 

Arango-Lasprilla JC, Cuetos F, Valencia C, Uribe C, Lopera F. (2007). 

Cognitive changes in the preclinical phase of familial Alzheimer's 

disease. J Clin Exp Neuropsychol. 29, 892-900. 

Devanand DP, Pradhaban G, Liu X, Khandji A, De Santi S, Segal S, 

Rusinek H, Pelton GH, Honig LS, Mayeux R, Stern Y, Tabert MH, de Leon 

MJ. (2007). Hippocampal and entorhinal atrophy in mild cognitive 

impairment: prediction of Alzheimer disease. Neurology, 68, 828-36. 

Ding B, Chen KM, Ling HW, Sun F, Li X, Wan T, Chai WM, Zhang H, Zhan 

Y, Guan YJ. (2009). Correlation of iron in the hippocampus with MMSE in






patients with Alzheimer's disease. J Magn Reson Imaging, 29, 793-8. 

McGeown, W.J., Shanks, M.F., Forbes-McKay, K.E., Venneri, A. (2009). 

Patterns of brain activity during a semantic task differentiate normal 

aging from early Alzheimer's disease. Psychiatry Res., 173, 218-27. 

“CORRELATI FUNZIONALI E STRUTTURALI DELLE FUNZIONI COGNITIVE 

NELLA FASE PRE-CLINICA E CONCLAMATA DELLE DEMENZE 

DEGENERATIVE” 

Cassa di Risparmio di Parma e Piacenza 

24 mesi 

The present project aims to investigate the neurobiological substrates 

associated with the insurgence and progression of Alzheimer’s disease 

(AD). In particular, the attention will be focused on the study of brain 

regions more vulnerable and affected by AD pathology and responsible 

for AD symptoms, such as hippocampal, posterior cingulate and parietal 

circuits involved in memory function. The project will concentrate on the 

identification of distinct neurofunctional and structural pattern which 

characterize the different forms of dementia, in order to establish the 

effectiveness of neuroimaging techniques in the diagnostic process of 

dementia. Particular attention will be give on the study of neuroimaging 

correlates of Mild Cognitive Impairment as this is considered a critical 

phase in the progression to dementia: the identification of typical preclinical 

neuroimaging markers of AD will provide the possibility to act an 

early and effective intervention.

Nome Stefano F. Cappa 

Contatti 

Tel. 

E mail 

DIBIT via Olgettina 58 

20132 Milano tel 0226434887 

cappa.stefano@hsr.it 

Istituto/Dipartimento Vita Salute University and 

San Raffaele Scientific Institute, Division of Neuroscience 

Proposta di ricerca. 

Our multi-disciplinary reaserach unit, combines expertises from different backgrounds (neurology, 

neuropsychology, linguistics, cognitive psychology, epistemology), with the general aim to investigate the 

neural mechanisms of language and high-order cognition. The experimental approaches include 

behavioural studies in normal subjects and in neurological patients, brain imaging using magnetic 

resonance techniques and positron emission tomography, and neurophysiological investigations based on 

evoked responses and transcranial magnetic stimulation. In the field of dementia we are leading a number 

of investigation in the areas of linguistic function, memory and social neuroscience. Besides the theoretical 

interest , these studies are of relevance for issues such as early diagnosis (in particular distinction from 

healthy aging), differential diagnosis among different causes of degenerative dementia, and assessment of 

treatment effect. The studies are typically based on the development of innovative behavioural testing 

procedures, combined with multimodal imaging, biomarkers and genetic investigations, and are largely 

based on collaboration amiong multiple clinical and research units, both within the San Raffaele 

Foundation and with other national and international institution. 

Area di interesse identificata Cognitive neuroscience of dementia: clinical and theoretical aspects 






The reseach has been supported by grants from the Ministry of Health, 

the MIUR, the EEC and Human Frontiers organization

Nome Daniela Perani 

Contatti 

Tel. 

E mail 

02-26432224 -2223 

daniela.perani@hsr.it 

Istituto/Dipartimento V-S San Raffaele University, Division of Neuroscienze and Nuclear 

Medicine Dpt. San Raffaele Scientific Institute 


Area di interesse identificata Studies are aimed at investigating with PET the brain functional 

parameters and neurotransmission systems in neurological and 

psychiatric diseases : 



18 F-FDG is used for the evaluation of brain glucose metabolism and 11 C - 

PIB for amyloid deposition. Present protocols include dementias (AD and 

FTLD, LBD, PDD) in early and pre-clinical phase, including in Mild Cognitive 

Impairment. 

11 C-PK and its developed forms are used in the study of 

neuroinflammation. Present protocols include Parkinson’s disease and 

parkinsonisms such as Lewy Body Dementia, prion diseases, and 

Amiotrophic Lateral Sclerosis. To this aim mathematical models for 

quantification of activity are under development. 

11 C-Raclopride and 11 C-Fe CIT for the study of dopaminergic system. 

Present protocols include Parkinson’s disease and parkinsonisms, 

particularly in early phase. Voxel-based methods for automatic 

quantification are under study validation. 

11 C MP4 for measuring the AchE activity, in AD and MCI subjects in order 

to obtain correlations between reduction of enzymatic activity and 

cognitive status. To this aim mathematical models for quantification of 

activity are under development. 

11 C-MDL for 5HT2 serotonin receptors and 11 C-raclopride. Present 

protocols include psychiatric conditions such as obsessive compulsive 

disorders and depression 

11 C-FMZ for the evaluation of the gabaergic system. Present protocols 

include movement disorders such as dystonias (genetic and sporadic 

cases) 

1. Diagnosis by Statistical and Intelligent Systems (ICT 2009, 5.3). 

2. Molecular imaging for the early diagnosis and monitoring of 

Alzheimer’s disease in old individuals with cognitive disturbances:




an ADNI-compatible prospective study. 

3. Identification of diagnostic biomarkers in Amyotrophic Lateral 

Sclerosis patients, and development of related computational 

methods. 

1. Comunità Europea 

2. Ministero della Sanità 

3. Ministero della Sanità 

1. 2010-2013 

2. 2010-2012 

3. 2008-2010 

1. Validazione di nuovi metodi voxel-based e pattern recognition 

per il neuroimaging funzionale PET, SPECT e strutturale MRI nella 

diagnosi precoce e nella diagnosi differenziale delle malattie 

neuroddegenerative associate a demenza. 

2. Studio dei correlati neurofunzionali, anatomici e molecolari nel 

disturbi cognitivi lievi. 

3. Studio di biomarker in vitro e in vivo nella Sclerosi Laterale 

Amiotrofica.

Nome Maurizio Popoli 

Contatti 

Tel. 

E mail 

Tel: +390250318361 

E-mail: maurizio.popoli@unimi.it 

Istituto/Dipartimento Center of Neuropharmacology, Department of Pharmacological Sciences, 

CEND, University of Milan 


Area di interesse identificata 3C. Global proteomics of human fibroblasts for the identification of 

early biomarkers for dementia and Alzheimer’s Disease 










Identification of biomarkers for Alzheimer’s dementia: genomics and 

proteomics of neurodegeneration 


2004-2006 

Main object of the present project was the identification of genetic and 

biological markers for Mild Cognitive Impairment (MCI) in order to 

identify a strategy useful for an early diagnosis and for the individuation 

of subjects at risk for the development of Alzheimer’s disease. 

Neurodegeneration, physiological cerebral aging and dementia: an 

integrated approach for a early diagnosis, evolution predictors, and for an 

advanced care organization. 

Ministero della Sanità 

2000-2003 

Aim of this project was the investigation of biological, cellular, genetic 

and neurotoxic processes underling dementia for the identification and 

definition of predictive markers of pathology in the general population 

and in presence of risk factors.

Nome Gianfranco Spalletta 

Contatti 

Tel. 

E mail 

Via Ardeatina, 306 -00179 Rome 

06-51501575 


Istituto/Dipartimento IRCCS Santa Lucia Foundation/Department of Clinical and behavioral 

neurology - Neuropsychiatry laboratory 


Area di interesse identificata Parkinson disease and parkinsonisms 






Analysis of cognitive and neuropsychiatric disturbances in Parkinson 

disease and parkinsonisms: implications for early diagnosis 


22/12/2008 - 22/12/2011 

The prevalence rate for neuropsychiatric symptoms of depression, 

anxiety, and hallucinations in subjects with Parkinson Disease (PD) has a 

very wide range. This suggests how undefined the results of studies done 

up till now are. Moreover, longitudinal studies suggest that the majority 

of subjects with PD could develop dementia during the course of the 

disease. This study aims at evaluating the differential features of 

cognitive and neuropsychiatric disturbances in subjects with PD and other 

Parkinsonisms at the onset of the disease vs. advanced phases. A crosssectional 

model could help to understand the phenomena changing along 

time in a time-limited project. If psychopathological disturbances 

associated with Parkinson disease are difficult to be studied, those that 

accompany other motor disturbances are almost completely unknown. 

Therefore, it is urgent to fill this gap in knowledge with studies aimed at a 

better diagnostic typing between PD, LBD and other degenerative and 

vascular parkinsonisms. Another important issue concerns the validity of 

diagnostic criteria of DSM-IV for mood disturbances and anxiety 

disturbances in subjects with motor disturbances. Thus, in this crosssectional 

study, 300 patients affected by motor disturbances will be 

recruited. In particular, we will include 150 subjects with idiopatic 

Parkinson Disease (PD) of which 50 at onset and 100 in the advanced 

phase, 50 subjects with Dementia with Lewy Bodies (DLB); 50 subjects 

with vascular PD; 25 subjects with Progressive Supranuclear Palsy (PSP); 

20 subjects with corticobasal degeneration (CBD) and 10 subjects with 

multisystemic atrophy (MSA). Patients will be assessed using a clinical 

battery comprising: a neurological battery including socio-demographic 

and clinical history, Unified Parkinson’s Disease Rating Scale (UPDRS), and 

Hoehn and Yahr scale. The neuropsychological battery will include: Mini 

Mental State Examination, Rey 15-word test, reduced Wisconsin Card

Sorting, reduced Stroop Color-Word test, Verbal and visual-spatial Nback, 

Verbal semantic fluency, Verbal phonological fluency, Sentence 

construction, Immediate and deferred copy of Rey figure. The 

neuropschiatric battery will include: Psychiatric diagnosis according with 

the DSMIV-TR (in this case we will use both the standard diagnostic 

criteria and modified diagnostic criteria proposed by the Starkstein group 

in order to improve the neuropsychiatric diagnosis in patients with motor 

disturbances), Beck Depression Inventory, Toronto Alexithymia Scale, 

Penn Emotional Recognition Test, Snaith-Hamilton Pleasure Scale. In 

another part of the study we will include 20 idiopatic PD patients treated 

with Deep Brain Stimulation (DBS). They will be clinically evaluated with 

UPDRS Part III and with the neuropsychological and neuropsychiatric 

battery above-mentioned. The evaluation of subjects with stimulator 

implants will be carried out without any treatment and in 3 different 

phases: 1) during proper and stable antiparkinsonian therapy and 

Stimulator on, 2) during pharmacological therapy and DBS stimulator off 

since 90 minutes, 3) absence of pharmacological therapy with DBS 

stimulator on. The neuropsychological and behavioural tests will be 

carried out in a random way on different days. In the subgroup of 

patients with DBS the evaluation for depression will be carried out with a 

visual-analogical scale to instantly capture the severity of the depression 

in each of the 4 experimental conditions. Results of studies in this field 

will allow to propose new diagnostic criteria for neuropsychiatric and 

behavioural disturbances which will help clinicians in the early and 

differential diagnosis of movement disorders allowing also a better 

treatment. The research on patients in treatment with DBS will allow a 

better understanding of the features of non motor symptoms and 

directions towards treatments. The study will also provide a picture of the 

differential expression of cognitive versus neuropsychiatric symptoms 

within each pathological entity. Such achievements will be possibly imply 

a reduction of the burden caused by these diseases on the patient’s 

family, as well as a decrease of the social and heath costs.


Contatti 

Tel. 

E mail 

Via Ardeatina, 306 - 00179 Rome 

O6-51501575 





Area di interesse identificata Mild Cognitive Impairment and Dementias 






Cognitive and behavioural indicators of conversion from Mild Cognitive 

Impairment to neurodegenerative dementia and development of 

cognitive rehabilitation protocols 


01/01/2009 - 31/12/2011 

According to a four-group classification, the MCI amnestic single domain 

is characterized by a selective deficit of amnesic performances, multidomain 

amnestic MCI presents memory impairment and at least one 

other cognitive domain impairment, nonamnestic single domain MCI is 

associated with the compromise of one nonamnestic domain, and 

nonamnestic multi-domain MCI is associated with the compromise of two 

or more nonamnestic cognitive domains. The hypothetic usefulness of 

subdividing MCI to identify specifically the type of dementia to which the 

patient will convert has partially failed. However, there is solid evidence 

that multi-domain MCI has the highest rate of conversion in dementia. 

Although there is convergence on the description of the high risk of 

developing AD in subjects with chronic depression, often associated with 

hippocampal atrophy, there are also studies which clarify with sufficient 

reliability that other neuropsychiatric disturbances may predict 

conversion to AD. Regarding the fact that therapeutic strategies for the 

pathogenesis of AD have failed up till now, and considering the high risk 

of patients with MCI to develop AD, a revision of the clinical-diagnostic 

criteria of AD has been recently proposed. This revision makes AD 

diagnosis similar to those of amnesic MCI with the exception of biological 

and neuroradiological markers recognized as specific for AD diagnostic 

purposes. Unfortunately, the specific neuropsychiatric symptoms of AD 

were not included even in this recent diagnostic revision proposal. This 

research project will attempt to identify clinical characteristics useful for 

predicting the risk of developing neurodegenerative dementia from the 4 

different forms of MCI, the type of dementia more frequently associated 

with each clinical characteristics, the progression severity of the illness; d)

the effect of caregiver support on the illness progression, and the efficacy 

of cognitive rehabilitation on every subtype of clinical characteristics. The 

method of open study on cognitive rehabilitation will give an early clinical 

indication of the potential of this treatment. One hundred patients 

diagnosed with MCI will be included in the study. The patients included 

will carry out an in depth anamnestic evaluation and a 

diagnostic\categorical evaluation for MCI subtype. In addition, categorical 

neuropsychiatric disorders of depression, psychosis, and apathy will be 

assessed. Dimensional neuropsychiatric symptom severity will be 

measured with a battery of tests including: Neuropsychiatric Inventory-12 

items, CERAD Disforia, and Dementia Apathy Interview and Rating. The 

age at onset of each behavioural disturbance and the first memory deficit 

will be accurately evaluated. Moreover, To obtain a global index of 

cognitive impairment, we will administer the MMSE. The Mental 

Deterioration Battery and other cognitive task will be used to assess 

individual cognitive domains. A subsample of 30 MCI patients will be 

treated with 12 weeks cognitive rehabilitation. Also, a subsample of 

caregivers of 30 MCI patients will undergo psychometric evaluation at 

baseline and at the three and six-month follow-up. All subjects will be 

included during the first 6 months period of the project. All diagnostic, 

cognitive and neuropsychiatric tests will be administered every six 

months until 18 months from the first evaluation. 

This project will allow a more correct clinical-diagnostic subclassification 

of MCI subgroups in order to make the clinical subtypes of MCI more 

homogeneous both on the diagnostic level and the biological correlates. 

In particular, this most reliable subclassification should help to fasten the 

recognition of the disorders and the differential diagnosis, thus 

facilitating a more precocious and specific treatment of patients who will 

develop the different types of dementia. This project will also suggest the 

potential role of cognitive rehabilitation therapy and caregivers support 

on the cognitive and behavioural symptom progression of MCI. To obtain 

this result potentially means a reduced cost of the management of 

dementia for the national health service and a better outcome of the 

illness for patients.

Nome Paolo Calabresi 

Contatti Clinica Neurologica, Università degli Studi di Perugia, Ospedale S. Maria 

della Misericordia, 06132 Perugia, Italy 

e-mail: calabre@unipg.it 

Istituto/Dipartimento Clinica Neurologica, Laboratori di Neurologia sperimentale, 

Dipartimento di Specialitá Medico Chirurgiche e Sanitá Pubblica, 

Università degli Studi di Perugia 


Area di interesse identificata Electrophysiological study of striatal physiology in normal conditions 

and in experimental parkinsonism: use of genetic and pathogenetic 

animal models. 

Pre-clinical research is conducted at the Laboratory of Experimental 

Neurology of the Clinica Neurologica at the Hospital S. Maria della 

Misericordia in Perugia. The activities focus on the study of important 

neurological pathologies such Parkinson’s disease (PD) and other 

neurological and neurodegenerative conditions (Huntington disease, 

Alzheimer disease, Multiple sclerosis, stroke, epilepsy) by means of in 

vitro animal models. 

The main topic of the laboratory is the study of the mechanisms 

involved in the physiopathology of such diseases by means of up to 

date electrophysiology and behavioural techniques in tight 

collaboration with the Laboratories of Neurophysiology located at the 

Fondazione S. Lucia in Rome. 

Motor and non-motor symptoms occurrence in PD represent the 

downstream effect of a pathological cascade resulting in the 

degeneration of dopaminergic neurons of the substantia nigra pars 

compacta (SNpc) projecting to the nucleus striatum. In this nucleus 

physiological activation of neuronal pathways are able to produce longterm 

depression (LTD), long-term potentiation (LTP) and 

“depotentiation” of synaptic transmission that are all disrupted by the 

PD-related pathological process. While Levodopa (L-DOPA) remains the 

gold standard of symptomatic therapy of PD it also causes severe longterm 

side effects (L-DOPA-induced dyskinesia, LID). The cellular and 

molecular mechanisms of LID formation and maintenance have started 

to be elucidated. 

For the study of these phenomena we take advantage of several animal 

models of PD. A well accepted model is obtained with the injection of 6hydroxydopamine 

(6-OHDA) into the SN causing permanent DAdenervation 

of the ipsilateral striatum mimicking the symptoms of the 

human PD. The in vitro model of PD by acute application of the 

neurotoxin rotenone that affects the complex I of mitochondrial chain 

is an important tool to explore mitochondrial function linked to PD. In 

order to assess specific aspects of the pathogenesis of PD, we employ



Ente finanziatore Prin 2008 



transgenic animal lines expressing different mutant forms of αsynucleins 

such as A53T-α-synuclein mice and mice expressing human 

α-synuclein lacking the C-terminal 20 amino acids (α -Syn120). 

Furthermore, in order to study and to prevent LID development 

(priming) and maintenance in PD models we focussed on the 

modulation of the Ras-ERK pathway by the use of lentiviral vector (LV)mediated 

expression of either small harpin RNAs (shRNAs) specific for 

Ras-GRF1 and DARPP-32. 

Nuovi approcci terapeutici per le discinesie indotte dal trattamento con 

L-DOPA in un modello sperimentale di Parkinson: ruolo delle subunità 

del recettore NMDA e della via molecolare Ras-ERK 

LIDYAS - An innovative approach to treat levodopa-induced dyskinesia 

based on targeting striatal intracellular signalling 

Ente finanziatore Progetto San Paolo di Torino - Coordinatore: Istituto Nazionale di 

Neuroscienze (INN) 



A multidisciplinary approach to test the therapeutic potential of neural 

stem cell transplantation in preclinical mouse models of Parkinson’s 

disease 

Ente finanziatore Progetto Strategico ex art56 Centro San Raffaele del Monte Tabor 



Ente finanziatore Progetto Finalizzato 2005 


Meccanismi di danno neuronale alla base della neurodegenerazione 

nelle patologie dei gangli della base 

Titolo progetto SYNSCAFF - Synaptic scaffolding proteins orchestrating cortical synapse 

organisation during development. 

Ente finanziatore SIXTH FRAMEWORK PROGRAMME - PRIORITY [LSH-2003-2.1.3-5] 

[Cortical development] Proposal/Contract no.:511995 

Durata progetto Four years 


Mechanisms of L-DOPA-induced dyskinesia in an experimental model of 

Parkinson's Disease: focus on NMDA receptors 

Ente finanziatore Ministero dell’Istruzione dell’Università e della Ricerca (Prin 2005) 



Ente finanziatore Sigma Tau 

Durata progetto One years 

Ruolo dell’adenosina nella trasmissione sinaptica eccitatoria striatale: 

interazione con il sistema dopaminergico


Mechanisms underlying cell-type specific vulnerability of striatal 

neurons: implications for huntington's disease 

Ente finanziatore Telethon GGP02035 


Titolo progetto REPLACES - Restorative Plasticity At Corticostriatal Excitatory Synapses 

Ente finanziatore Comunità Europea - HEALTH-2007-2.2.1-7 - Grant agreement no.: 

222918 

Durata progetto Four years 

Titolo progetto Imaging and biological markers of disease progression 

Ente finanziatore Progetto STRATEGICO Ministero della Sanità - Sottoprogetto 5 

Durata progetto Two years

Nome Lucilla Parnetti 

Contatti 

Clinica Neurologica, Università degli Studi di Perugia, Ospedale S. Maria della 

Misericordia, 06132 Perugia, Italy 

Tel: +39 075 578 3545; Fax: +39 075 578 3621. 

Tel. 

parnetti@unipg.it 

E mail 

Istituto/Dipartimento Clinica Neurologica, Centro Disturbi della Memoria – Unità Valutativa 

Alzheimer - Laboratorio di Neurochimica Clinica, Dipartimento di 

Specialitá Medico Chirurgiche e Sanitá Pubblica, Università degli Studi di 

Perugia 


Area di interesse identificata Cerebrospinal fluid biomarkers in Alzheimer’s disease and other 


Clinical research in the Laboratory of Neurochemistry is performed in 

close contact with the Center for Memory disturbancies, the Center for 

Movement disorders and the Center for Demyelinating disease of the 

Hospital S. Maria della Misericordia in Perugia. In the Laboratory of 

Neurochemistry, a biobank of cerebrospinal fluid (CSF) and plasma 

samples from patients with neurological diseases is available, making 

feasible retrospective studies on several pathologies such as Alzheimer’s 

disease (AD), Parkinson’s disease (PD) and other neurodegenerative 

diseases. 

The main topic of the laboratory is the study of CSF biomarkers for the 

early and/or differential diagnosis of AD. The classical biomarkers Aβ1-42, 

tau protein (t-tau), and phosphorylated tau 181 (p-tau) are routinely 

assayed in CSF of patients with suspected dementia and/or patients with 

mild cognitive impairment (MCI) for which is also available a clinical 

follow up by means of neuropsychological tests.Techniques available in 

the Laboratory of Neurochemistry include mainly immunoassays such as 

ELISA and xMAP technique. The latter allows us to multiplex several 

analytes increasing the number of molecules that can be measured 

starting from the same volume of sample. 

CSF biomarkers show promise as a diagnostic tool in patients with AD, but 

recent multicenter studies showed that the levels of these biomarkers 

vary between different research centers. To study the source of variation 

our laboratory is actually involved in a world quality control program run 

by the Alzheimer Association in conjunction with Clinical Neurochemistry 

Laboratory in Gothenburg, Sweden, with the final goal of standardize 

these assays for clinical routine. 

Beside the classical CSF biomarkers we are actually studying other 

proteins as possible biomarkers for neurodegenerative diseases. We have 

recently shown that heart-fatty acid binding protein (H-FABP) is 

significantly increased in MCI patients which converted to AD and it may 

represent a new marker of neurodegeneration. Other projects are related 

to progranulin measurement in CSF, a protein which has been related to 

frontotemporal dementia and amyotrophic lateral sclerosis and to the 

evaluation of Aβ1-40 as a useful marker to differentiate various dementia 

conditions. Finally we are also evaluating the usefulness of the 

measurement of α-synuclein and classical CSF biomarkers to distinguish










synucleinopathies (PD and dementia with Lewy Bodies) from other 


cNEUPRO (contract no. 199 LSHM-CT-2007-037950) 

EU – FP6 

Three years (2007-2010) 

Recent research has clearly demonstrated that multiparametric 

neurochemical dementia diagnostics (NDD) in cerebrospinal fluid (CSF) 

does improve the early and differential diagnosis of dementias. cNEUPRO 

will apply advanced proteomic tools to discover novel neurochemical 

dementia markers in blood and CSF for the improved early and possibly 

predictive diagnosis of AD. A predictive dementia diagnosis will support 

the most effective use of forthcoming preventive therapeutic strategies. 

Our initiative will establish European SOPs for current neurochemical 

dementia diagnostics (NDD). A strong methodological impact is put on 

the quality of pre-analytical sample handling and clinical phenotyping, 

which has been neglected in industry-driven discovery studies. cNEUPRO 

integrates innovative biotech and bioinformatic companies with leading 

clinical and proteomic dementia research centres. cNEUPRO will also 

support the discovery of new diagnostic targets and is also promising to 

identify novel scaffolds for advanced molecular neuroimaging. 

CSF lysosomal hydrolases’ activity as possible marker of Parkinson’s 

disease 

MJFF 

1 year 

Objective/Rationale: 

Recent studies have linked lysosomal dysfunction to the accumulation of 

α-synuclein oligomers and α-synuclein-mediated cell death. Clinical, 

neuropathological and genetic associations between Gaucher’s disease, a 

lysosomal storage disease, and Parkison’s disease (PD) have been 

reported. Also, a reduction of the activities of β-glucocerebrosidase, α 

and β-mannosidase has been reported in CSF of PD patients. 

To date there is no accepted diagnostic test for Parkinson’s disease based 

on biochemical analysis of blood or cerebrospinal fluid (CSF). The 

potential use of CSF lysosomal enzyme activities as a diagnostic indicator, 

or as a marker of disease progression in PD, will be investigated. 

Moreover the CSF levels of alpha-synuclein – a candidate diagnostic 

biomarker of PD – will be combined with the lysosomal enzyme activities 

in order to verify if there is any association between these biochemical 

parameters.Project Description: De-novo and treated patients referring 

to the Section of Neurology, University of Perugia, Italy, will be included 

in the study. CSF samples will be collected from patients with PD (n=80) 

and healthy subjects (n=50). Lysosomal enzyme activities (betaglucocerebrosidase, 

alpha-mannosidase, beta-mannosidase, beta-

hexosaminidase, beta-galactosidase, alpha-fucosidase, arylsulfatase A, 

arylsulfatase B, cathepsin D, and cathepsin S) will be determined in CSF 

samples using specific fluorimetric substrates. The levels of monomeric 

and oligomeric alpha-synuclein, using a sensitive and specific ELISA 

method able to reveal levels as low as 1 pg/mL in human biological fluids, 

including plasma and CSF, will be also evaluated.


D. Diagnosis: biomarkers

Nome 

Roberta Ghidoni, Giuliano Binetti 

Contatti 

+39-030-3501725 

rghidoni@fatebenefratelli.it 

Istituto/Dipartimento IRCCS “Centro San Giovanni di Dio-Fatebenefratelli” 

Proteomics Unit, NeuroioGenLab Memory Clinic, Brescia, Italy 


Over the past 10 years, frontotemporal lobar degeneration (FTLD) has emerged as the most common cause 

of dementia under the age of 60 years, outstripping even Alzheimer disease in prevalence. The increasingly 

common occurrence of FTLD, and its devasting impact on families and patients within their most 

productive years, urges novel approaches to cure or to prevent it. To date, there are no Food and Drug 

Administration approved medications for FTLD. FTLD has a strong genetic component, with up to 50% of 

cases being caused by mutation. Recently the progranulin gene (PGRN) was shown to be the most frequent 

genetic determinant: 68 mutations have been described in 212 families which account for 5–10% of FTLD 

cases worldwide (http://www.molgen.ua.ac.be/FTDmutations/). At present, the most common PGRN 

mutations worldwide are the Arg493X mutation, which has been identified in 37 patients belonging to 30 

genealogically unrelated families with FTLD (the “Mayo Clinic” cohort) (Rademakers R et al,Lancet Neurol. 

2007) and the PGRN Leu271LeufsX10 mutation, that we identified in 49 patients belonging to 38 unrelated 

Italian families (the “Brescia” cohort) (Benussi et al. NBA 2008;Benussi L and Ghidoni R et al, NBD 2009; 

Benussi L et al. ADAD in press).We recently reported that low plasma progranulin levels predict progranulin 

mutations in frontotemporal lobar degeneration. (Ghidoni R et al, Neurology 2008). Progranulin therefore 

has become a promising target for new therapeutic FTLD approaches. Our resources are: 

1) Clinical expertise in FTLD: the IRCCS in Brescia has become a reference center for FTLD and related 

disorders serving a large geographical area in Lombardia. 2) The largest (along with the “Mayo Clinic 

cohort”) biological sample collection of PGRN mutation carriers (the “Brescia” cohort). Our scientific work 

in the last years has generated one of the largest cohort of FTLD families and biological sample collection of 

PGRN mutation carriers (affected and presymptomatic), that is an invaluable resource for research studies 

3) Cellular and animal disease models. 

Our specific aims are: 

1) Elucidating the role of progranulin in the disease onset and progression: we will study the molecular 

mechanisms underlying neurodegeneration in PGRN mutations carriers (plasma/serum) as well as in 

cellular (human fibroblasts and lymphocytes ) and animal models. 

2) Biomarkers discovery for the differential diagnosis of FTLD vs Alzheimer’s disease (AD). FTLD is often 

misdiagnosed as another type of dementia (AD) and hence biochemical diagnostic markers would aid in 

differential diagnosis. Analysis of CSF is so far the most convenient method for studying the biology of 

neurodegenerative diseases in living patients. For a global proteomic study, surface-enhanced laser 

desorption/ionization mass spectrometry (SELDI-TOF MS) as well as two-dimensional gel electrophoresis 

(2-DE) will be used to study differential CSF protein expression. 

3) Searching for new genes: Identification of novel disease-associated loci by linkage and gene expression 

analysis. 

Centers collaborating to this project: IRCCS Centro S.Giovanni di Dio-Fatebenefratelli, Brescia;. Istituto 

di Ricerche Farmacologiche "Mario Negri" , Milan (G. Forloni); Università degli Studi di Brescia (PF 

Spano); University of Milan (D. Galimberti, E. Scarpini); IRCCS Besta, Milan (F. Tagliavini) 

Area di interesse identificata Translational research 



Proteomics of cognitive and movement disorders: Identification of 

molecular mechanisms associated with disease onset and phenotypic 

variability of frontotemporal lobar degeneration- (2009-2633)








FONDAZIONE CARIPLO, Bando Ricerca Biomedica 2009 

01-01-2010/01-07-2012 

The main objective of the present study is to unravel the 

pathophysiological bases of progranulopathies by 

1) proteomic approach- For a global proteomic study, SELDI-TOF as well 

as two-dimensional gel electrophoresis (2-DE) will be used to study 

differential protein expression in subjects carrying or not PGRN mutations 

2) neuroimaging study- The aim of the neuroimaging study is to describe 

the MR features of PGRN mutations with conventional, non-conventional, 

and functional neuroimaging. 

3) personality and behavioral assessment- We will assess the personality 

and behavioral trait of asymptomatic at risk family members carrying 

PGRN mutations 

4) cellular and animal models studies- 

Project 1: “Validation of genetic and biochemical markers for early 

diagnosis of AD and the prediction of conversion of MCI into AD, and 


prognostic accuracy”. 

Project 2: “Marker biologici di neurodegenerazione utilizzabili nella 

pratica clinica ai fini della diagnosi precoce e differenziale, della 

individuazione delle forme con risposta ottimale alle attuali terapie” 

Project 1: Italian Ministry of Health, Progr. Strategico PS39, prog. 1 

Project 2: Italian Ministry of Health, Progr. Strategico conv.71, prog. 6 

Project 1: 01-01-2009/31-12-2010 

Project 2: 07-01-2008/07-04-2010 

These programs have two primary aims: 

1) The development and validation of a multi-factorial protocol that 

integrates molecular, imaging, neurophysiological, neuropsychological 

and behavioral data for early diagnosis of AD, in particular during the 

preclinical phase and the prodromal stage of "mild cognitive impairment" 

when the symptoms are not severe enough to fulfill the current 

diagnostic criteria for AD. 

2) Validation of this new diagnostic protocol within a regional public 

health network, and evaluation of the health care, organization and 

economic implications of its transfer to the National Health Service.

Referente: Prof. Carlo Ferrarese 

Phone. 039.233.3595 

Fax. 039.233.2449 

Email: carlo.ferrarese@unimib.it 

Clinica Neurologica, Ospedale San Gerardo 

Via Pergolesi 33 – 20052 Monza (MB) 

Laboratorio di Neurobiologia 

Dipartimento di Neuroscienze e Tecnologie Biomediche – Università di Milano-Bicocca 

Via Cadore 48 – 20052 Monza (MB) 

The main interest of our group consists in the integration of clinical and basic scientific research to 

understand the pathogenic mechanisms of neurodegenerative disorders and to identify new strategies for 

diagnosing, treating and preventing such untreatable diseases. Our center is composed by a clinical core 

(Department of Neurology, 42 beds plus Laboratory of Neurophysiology and Laboratory of 

Neuropsychology) at the san Gerardo Hospital (Monza, Italy) and a biological facility (Laboratory of 

Neurobiology, Department of Neuroscience and Biomedical Technologies, University of Milano-Bicocca). 

Regarding dementias and cognitive dysfunction, the main body of our clinical work is centered at the 

Memory Clinic (U.V.A. outpatient facility) that sees more than 300 patients among Alzheimer’s disease (AD) 

and other types of dementia. Clinical activity is deeply involved with research epidemiological, clinical, and 

neurobiological. Novel experimental therapies are offered; for example, a phase 3 trial testing the efficacy 

of an anti-beta amyloid antibody in AD is currently recruiting patients. 

The strict connection existing between the Memory Clinic and the Laboratory of Neurobiology allows 

studying novel peripheral markers in AD (mainly biochemical alterations in blood elements and skin 

fibroblasts) that might in the future be used for accelerating diagnostic processes, helping during follow-up, 

targeting personalized therapies, and refining prognostic previsions. Bioresearch methods currently 

employed include: cell cultures, immunochemistry, molecular biology, binding and uptake techniques, 

HPLC, confocal microscopy. For example, our group recently demonstrated an increase of sAPPα�plasma 

levels with respect to healthy controls, while no differences were found regarding beta-amyloid (Abeta) 

levels. Moreover, we set up an ELISA assay for the determination of plasma anti-Abeta 1-42 levels, although 

no differences were found between AD patients and controls. However, we observed that AD patients not 

receiving acetylcholinesterase medications (AChEI) displayed anti-Abeta 1-42 antibodies plasma levels 

significantly lower with respect both, healthy controls, and AD patients receiving AChEI. Hence, we are 

currently analyzing the effect of the AChEI therapy on the immune response in AD patients. A putative 

immunomodulatory effect of these drugs might eventually further support immunoglobulin-based 

therapies for treating dementia. Furthermore, our group extensively studies the interactions existing 

between glutamatergic system, APP processing and signal transduction mechanisms (kinases) in peripheral 

(platelets, fibroblasts, lymphomonocytes) and cell models (neuroblastoma SHSY5Y). Another 

neurobiological interest of our group is related to Abeta catabolism. We focused on neprilisin, a major 

Abeta catabolic enzyme, studying its activity and expression in human fibroblasts. Neprilisin activity is 

reduced of ~50% in AD versus controls, while the expression in unchanged. In vitro “aged” Abeta treatment 

reduces neprilisin expression in both groups, while the activity decreases in controls and increases in AD

patients, possibly due to a compensatory mechanism; this preliminary data suggests that Abeta is able to 

influence its own catabolism. A more innovative approach for reducing the toxic effect of Abeta consists in 

administering nanoparticles, molecules of nanometric dimensions able to bind Abeta limiting its 

aggregation and resulting toxicity. Within an FP7 European project we are testing the biocompatibility of 

various nanoparticles, and their ability to bind Abeta in vitro (cultured fibroblasts) and in biological fluids 

(plasma and serum) in order to identify the best candidates for future in vivo administrations. 

In the neuropsychological field, our current interests include: (a) development and validation of multiple 

ultrafast tests (5 minutes duration max) of the global cognitive status in order to propose them as rapid 

screening tools for general practitioners; (b) validation of a test of divided attention for the differential 

diagnosis of degenerative dementias; (c) development and validation of a novel test of denomination for 

the differential diagnosis between mild cognitive impairment (MCI) and dementia; (d) extensive study of 

visuo-spatial functions by a broad test battery, in order to define the differential cognitive profile between 

Lewy body dementia, corticobasal degeneration and posterior cortical atrophy; (e) study of the 

experimental neuropsychological (decision making, moral judgment, multi-tasking, humour appreciation) 

and neurobiological correlates of BDNF in AD, frontotemporal dementia and Lewy body or Parkinson 

dementia; (f) multicentric longitudinal cohort study recruiting patients affected by degenerative and 

vascular dementias or MCI to be followed for five years by repeatedly administering a neuropsychological 

test battery. 

Finally, our clinical department collaborates with the Department of Nuclear Medicine - Molecular 

Bioimaging Centre for both clinical and research purposes. Currently we are studying the existence of 

correlations between biochemical markers and functional neuroimaging data (PET-SPET). Tracers able to 

mark beta-amyloid or microglial activation, among others, might eventually integrate and confirm the value 

of the above described peripheral markers.

Nome Piero Parchi M.D., Ph.D. 

Contatti 

piero.parchi@unibo.it 

phone: 051-2092740 

fax: 051-2092751 

Istituto/Dipartimento Dipartimento di Scienze Neurologiche 


Aree di interesse identificate 





Abstract (estratti) del 

progetto 

Università di Bologna, Italy 


- Standardization of diagnostic criteria and diagnostic 


- Early diagnosis; Biomarkers 

- Basic research: Biochemical characterization of 

abnormal protein aggregates to understand phenotypic 

diversity in neurodegenerative diseases 

1) Molecular Pathology of the Human Brain; 

2) Development of new diagnostic approaches for prion 

diseases 

3) Validation and search of CSF biomarkers for the prediction 

of the clinical conversion from mild cognitive impairment 

(MCI) to dementia 

1) EU, 2) Italian Ministry of Health, 3) Galletti Foundation 

1) 5 years (Brain-Net II, project ended December 2009), COST 

application currently under evaluation (at final step). 

2) 3 years 

3) 3 years 

1) Studies on human brain tissue have enabled the discovery 

of key factors of the most common brain diseases. Aβ, tau, 

synuclein and TDP-43 are outstanding examples of proteins 

identified in diseases such as Alzheimer (AD), Parkinson (PD) 

and Frontotemporal Lobar Degeneration (FTLD). The advent of 

molecular genetic techniques enabling insights into temporal or

permanent changes of genetic activity (epigenetics) and the 

identification of novel regulators of translation (miRNA) have 

ushered in a new era that makes investigations of human brain 

tissue still indispensable. In this project ) we will provide high- 

quality human brain tissue as an essential research resource by 

applying and further developing a shared database, common 

diagnostic criteria and ethical standards at European level. 

Furthermore, new technologies will be employed to investigate 

the causes of neurodegenerative disorders using such tissues. 

This collaborative group of experts (see http://www.brainnet- 

europe.org) competent in all aspects of work with human brain 

tissue will foster European research on these devastating brain 

diseases. 

2) The overall objective of this project is to improve the 

diagnosis and the prognosis of human TSE diseases through the 

development of new biochemical tests and the optimization of 

already available diagnostic tools. To this aims we will apply 

new proteomic approaches to identify TSE-specific patterns in 

the CSF and in plasma (or serum) of patients with sporadic CJD 

and we will characterize novel biochemical properties of the 

abnormal PrP TSE aggregates that may further improve the 

strain-specific diagnosis of the disease subtype. 

3) As one of the Units involved in a larger project (see Form 

sent by Dr. Gallassi from our Department) we will validate and 

search for molecular CSF markers that might be of value in 

predicting the conversion from MCI to full-blown dementia and 

critical for the monitoring of patients once new therapies 

become available. We will focus on already established markers 

such as A-beta, total tau and fosfo-tau and more recently 

discovered molecules such as TDP-43 and progranulin. Different 

techniques such as Elisa and western blotting will be applied. 

The data will be correlated with those obtained from the 

neuroimaging and neuropsychological serial evaluation.

SICILIANO 

Analisi biochimica di parametri di stress ossidativo. 

Uno dei problemi clinici più attuali riguardala difficoltà a comprendere se lievi disturbi della memoria in 

età senile siano oppure no segni iniziali di Malattia di Alzheimer. A tal proposito va ricordato come 

attualmente si faccia riferimento a un quadro clinico-neurologico quale il decadimento cognitivo lieve 

(“Mild Cognitive Impairment o MCI”) ,quale condizione caratterizzata, come appunto il termine indica, da 

una lieve compromissione di funzioni mentali, in primo luogo la memoria, ma potenzialmente a rischio di 

sviluppare nel tempo una malattia di Alzheimer e pertanto da seguire attentamente nel suo ulteriore 

decorso. Un altro aspetto fondamentale è inoltre rappresentato dal fatto che, in assenza di un marcatore 

biologico in vivo che permetta di fare diagnosi di certezza della malattia in vita, assume particolare 

importanza la ricerca di parametri di laboratorio che siano in grado, in aggiunta a fornire elementi 

indicativi sulle cause di malattia, di essere indicatori di eventuale rischio, evoluzione e diagnosi precoce 

di malattia, oltre che predittivi di risposta al trattamento. 

Lo stress ossidativo è stato chiamato in causa come fattore implicato nella patogenesi delle malattie 

neurodegenerative, inclusa la malattia di Alzheimer, in base all’osservazione che si ha evidenza precoce 

di danno mediato da specie reattive dell’ossigeno, non soltanto a livello del sistema nervoso centrale ma 

anche a livello periferico. 

Poiché le specie reattive dell’ossigeno sono molecole altamente instabili, la loro misura diretta in 

campioni di plasma o siero può non riflettere la loro reale concentrazione in vivo; per tale motivo 

vengono dosati solo i prodotti finali di stress ossidativo. Distinguiamo i biomarker possono essere distinti 

in due grandi gruppi: 

1. Marker di danno ossidativo alle proteine, ai lipidi e al DNA, che rivelano lo stato di ossidazione di 

tali macromolecole e che danno un’idea del grado di gravità del processo degenerativo in atto all’interno 

della cellula; 

2. Marker dello stato antiossidante, che riflettono la capacità totale dei sistemi cellulari, enzimatici 

e non enzimatici, di difesa contro l’attacco delle specie chimiche reattive. 

Presso il nostro laboratorio sono valutati markers di danno ossidativo alle proteine (AOPP), markers dello 

stato antiossidante quali la capacità ferro-riducente del plasma (FRAP) e la concentrazione del glutatione 

totale, ridotto e ossidato. 

Gli AOPP consistono in un insieme di proteine tra cui la tiroglobulina, la γ-globulina, l’albumina e la 

mioglobulina (Witko-Sarsat et al., 1996). 

L’elettroforesi delle proteine mostra che il picco di AOPP ad alto peso molecolare è prevalentemente 

dovuto all’albumina che appare sotto forma di aggregati che probabilmente derivano da ponti disolfuro 

e/o da "cross-linking" della tirosina; al contrario il picco di AOPP a basso peso molecolare contiene 

albumina nella sua forma monomerica (Witko-Sarsat et al., 1996). In "vivo" i livelli plasmatici di AOPP 

correlano con i livelli di dimeri di tirosina, un marcatore di danno ossidativo delle proteine, e con la 

pentossidina, un prodotto di glicosilazione strettamente associato al danno ossidativo delle proteine 

(Selmeci et al., 2005). La relazione tra gli AOPP e i prodotti di perossidazione lipidica appare invece poco 

chiara. Le attuali conoscenze sembrano indicare che i lipidi non sono necessari per la formazione degli 

AOPP, ma che in “vivo” possono aumentare tale processo (Witko-Sarsat et al., 1996). 

Al valore della FRAP contribuiscono, per circa il 60% l’acido urico, per il 15% l’acido ascorbico, per il 5% 

l’α-tocoferolo, per il 10% le proteine e per il 5% la bilirubina (Benzie et al., 1996). La metodica che 

permette di determinare questo parametro sfrutta la capacità di questi antiossidanti di ridurre il ferro, - 

sotto forma di ione ferrico presente nel reattivo FRAP, - a ione ferroso. 

La principale funzione del glutatione è quella di operare come antiossidante proteggendo le cellule 

dall'azione di specie proossidanti (Dringer et al., 2000). Le proprietà antiossidanti del glutatione sono

dovute alla presenza del gruppo sulfidrilico della cisteina che gli permette di interagire sia con specie 

reattive dell’ossigeno, sia con altre sostanze elettrofile nell’ambito di numerosi sistemi antiossidanti e 

non (Pompella et al., 2003). Il glutatione allo stato ridotto (GSH) espleta la sua funzione riducendo i 

radicali liberi e convertendosi in glutatione ossidato (GSSG), composto da due molecole di glutatione 

unite da un ponte disolfuro. In questo senso, il GSH interviene nel metabolismo dell’acqua ossigenata, 

degli idroperossidi e di altri perossidi organici, è capace di inattivare il radicale idrossile, radicali 

perossilici, il perossinitrito, l’acido ipocloroso, radicali alcossilici e l’ossigeno singoletto. Può anche 

ripristinare la forma ridotta di altri antiossidanti come la vitamina E e la vitamina C. 

Valutazione clinica, test da sforzo e prelievi ematici per la valutazione dello stato ossidativo 

I parametri di stress ossidativo sopra descritti possono essere studiati sia in condizioni basali che dopo 

protocolli di esercizio. Le principali sedi di produzione di radicali liberi sono i mitocondri, gli organelli 

intracellulari deputati al metabolismo ossidativo; i mitocondri svolgono altresì un ruolo di primo piano 

nel mantenere l’omeostasi del calcio e nell’innescare la cascata di segnali che sottendono la 

degenerazione cellulare attraverso il meccanismo della apoptosi. 

Lo studio della funzione mitocondriale e dello stress ossidativo ad essa connesso può essere valutato 

attraverso l’andamento dei parametri ematici di stress ossidativo in relazione all’esercizio muscolare. 

Esempio di test da sforzo per la valutazione di parametri di stress ossidativo: 

I pazienti sono sottoposti a un test di sforzo incrementale sui muscoli dell’avambraccio, eseguito con 

l’ausilio di un miometro connesso ad un “hand-grip” (Digital Multi-Myometer, MIE Medical Research Ltd., 

Leeds, UK). Il protocollo di esercizio incrementale consiste nell’impugnare l’hand-grip del miometro e 

contrarre massimamente la mano contro la resistenza offerta dallo stesso, determinando, in Newton, il 

livello di contrazione volontaria massimale (CVM). Saranno eseguite, da tutti i pazienti, una serie di fasi 

contrattili o “steps”, condotte in maniera intermittente per un periodo di un minuto, con intervalli di 

riposo di 2 minuti tra uno “step” e l’altro. In ciascuno “step” le contrazioni intermittenti sono eseguite con 

frequenza di una al secondo contro la resistenza richiesta offerta dal miometro. L’esercizio inizia con un 

primo “step” al 10% della CVM, un secondo “step” al 40% della CVM ed un ultimo “step” al 70% della 

CVM. Il paziente potrà seguire su un display luminoso il livello di forza generato in ogni contrazione. 

Questo tipo di esercizio è principalmente aerobico all’inizio del test e diviene progressivamente 

anaerobico man mano che aumenta la forza esercitata per progressivo reclutamento delle unità motorie 

rapide (Milner-Brown et al., 1973). I prelievi ematici venosi per la determinazione dei marcatori 

biochimici di stress ossidativo sono eseguiti a livello basale, dopo il secondo “step”, dopo il terzo “step” e 

dopo 15 minuti dalla fine dell’esercizio.


Contatti 


Tel. +39 051 2097947 

Cell. +39 335 310979 




Multidisciplinary projects Cognitive decline in multiple sclerosis. Cognitive defects are 

recognized as early events in MS, not correlated with white matter 

lesion. We are interested in studying the pathogenesis of early 

neural distress and lesion in MS animal models and neuroprotective 

strategies 

Standardization of diagnostic 

criteria and diagnostic 

instruments, harmonization 

and assessment tools 






Quality control programs for Abeta fragments in CSF and plasma 

and tau/Ptau in CSF using xMAP platform. Focus on clinical trials 

standardization 

Translational medicine for novel diagnostic and therapeutic tools for 


Joint Regione Emilia Romagna-University of Bologna 

3 years, under negotiation 

Development of a GLP-grade service for drug testing in animal 

models of Alzheimer disease, based on computerized analysis of 

video-tracking for learning and memory performance. Development










of cell-based platform based on neural stem cells derived from 

Alzheimer mice for the screening of chemical libraries of anti- 

Alzheimer drugs. 

Effect of low-dose of ….. in chronic administration on cognitive 

behavior and morphological parameters in Tg2576 mouse 

Drug Company 

(covered by disclosure agreement) 

12 months 

The project is aimed to evaluate efficacy of a novel gammasecretase 

inhibitor on behavioral performance, histopathology, 

biomarkers and synapse morphology and molecular markers in mice 

curring the swedish mutation of APP. This esperiment will test 

efficacy of chronic treatment from 5 to 19 months of age. Two 

doses of the test compund and one dose of a reference will be 

tested in transgenic vs wild tipe, age matching mice 

Plasma assay of Ab40/Ab42 (multiplex X-MAP-based immuno assay 

technology) in phase 1 clinical trial 

Drug Company 

(covered by disclosure agreement) 

20 months 

Placebo-Controlled, Ascending Single-Dose Study to Evaluate the 

Safety, Pharmacokinetics and Pharmacodynamics of …. in Healthy 

Young Male Subjects. The safety and tolerability of five ascending 

oral doses of … will be evaluated in 6 separate study sessions. In 

each of the study sessions, subjects will be randomized to … or 

placebo treatment. Overall, 60 subjects will participate in the study, 

48 treated with … and 12 treated with placebo. Our lab in 

committed in efficacy variables blind analysis (Plasma �-amyloid 

concentrations, A40/A42 fragments). Joint QC initiative “round 

Robin” study, an international interlaboratory evaluation of AB 1- 

40/1-42 levels from a common plasma sample set, toward 

standardization of multiplex/multiparametric methods for AB 1- 

40/1-42 quantification in plasma. Participating centers, 15; number 

of samples to analyze, 19. 

Remyelination failure in multiple sclerosis: a case of inflammationinduced 

tissue hypothyroidism?



AISM/FISM 

Principla investigator: Luciana Giardino 

2 years 

Abstract del progetto This project is aimed to investigate a possible mechanims for 

remyelination failure in multiple sclerosis. This project is aimed to 

verify if the inflammation associated to demyelination causes a 

defect of T4 to T3 conversion or a decreased expression of thyroid 

hormone receptors (nuclear receptors and membrane transporters) 

in the nervous tissue, thus leading to a local/cellular hypothyroidism 

and, thus, to the defect of white matter repair. This project could 

also provide insides for other neurodegenerative disease including 

white matter alterations, like Alzheimer disease 


Contatti 


Tel. +39 051 2097947 

Cell. +39 335 310979 




Development of competitive 

animal models for AD 

Basic research 


GLP grade animal facility and behavioural testing for cognitive 

performance in AD animal models using video tracking and 

computer analysis; screening of new molecules after acute 

administration 

Brain metabolism of thyroid hormone: a risk factor for AD? We are 

interested in exploring possible connections between thyroid 

hormone content, receptor and activating enzyme expression, APP 

metabolism and Ab toxicity in the brain in normal and AD mice. 

Recent advances in knowledge on thyroid hormone brain 

metabolism and molecular biology of nuclear and membrane 

thyroid hormone receptors and transporters offer new possible 

targets to explore this topic.

Nome Fabrizio Tagliavini 

Contatti 

Tel +39 02 2394 2260; Fax +39 02 2394 2101 

Email: ftagliavini@istituto-besta.it 

Sito web: http://www.istituto-besta.it 

Istituto/Dipartimento UO Neuropatologia – Neurologia 5 


Via Celoria ,11 - 20133 Milano- ITALY 


The research group: The Division comprises (i) a Clinical Unit (Dementia Center) devoted to the diagnosis 

and treatment of patients with degenerative dementias (over 2000 out-patients and 150 in-patients per 

year) and (ii) a Laboratory Unit (certified ISO 9001:2000 - Registration Number 26080, dedicated to the 

analysis of genes and biomarkers associated with degenerative dementias, post-mortem characterization 

of the disease process and disease-specific protein, and experimental studies on disease pathogenesis and 

the development of therapeutic strategies. The course of action of this activity can be expressed as a “Bed 

to Bench to Bed” cycle, with the ultimate goal to identify disease-modifying drugs in preclinical settings 

and apply them to patients. In this regard, the Clinical Unit is currently coordinating a multicentre phase II 

clinical trial supported by AIFA, to test the effectiveness of an anti-amyloidogenic molecule identified in 

experimental models. This comprehensive approach has also the great advantage to collect biological 

samples (plasma, DNA, CSF and brain tissue) from fully characterized patients. In this regard the 

Laboratory Unit is part of the Network of Excellence “BrainNet” Europe devoted to biobanking, 

harmonization of assessment tools and standardization of diagnostic criteria of neurodegenerative 

diseases. 


comprises 13 post-doctoral fellows (7 PhD, 3 MDV, 1 MD, 1 psychologist), 2 PhD students and 1 technician 

who are committed to the research activities on degenerative dementias with different expertise and 

roles. On the overall, the team has large experience in a variety of techniques spanning from 

neuropathology (including immunohistochemistry, morphometry, electron microscopy and atomic force 

microscopy), to molecular genetics, biochemistry (purification and characterization of disease-specific 

proteins), cellular and molecular biology, and animal models. The Division of Neuropathology is provided 

with fully equipped laboratories for histology, immunohistochemistry, electron microscopy, atomic force 

microscopy, biochemistry, and molecular and cellular biology, and a 60 sqm BL3 facility. Moreover an 

animal facility composed by a 140 sqm environmental controlled area, provided with a HVAC system and 

heap-filtered ventilated racks, with separate rooms for maintaining and breeding mouse lines, and 

carrying out surgical procedures, collection of samples and post-mortem examination is available. 

Research lines on Alzheimer’s disease 

A. Clinical research 

• Early diagnosis of Alzheimer’s disease (AD) 

We will characterize the phenotypic expression of AD and other degenerative dementias through 

multiplex analysis of a number of variables including neuropsychological and behavioural profile, 

neuroimaging, neurophysiological changes with special attention to sleep abnormalities, genetics, and 

plasma and CSF biomarkers. Furthermore, we will carry out longitudinal studies of presymptomatic 

carriers of gene mutations to detect early markers of conversion to the disease state. 

• Clinical trials 

We will carry out phase II and phase III clinical trials to assess the effectiveness of potentially diseasemodifying 

drugs, including molecules targeted to Aβ and protein tau. 

• Identification of disease-specific biomarkers 

We are currently working on a peripheral marker of AD having high specificity and sensitivity, based on 

cyclic amplification of misfolded Aβ and Aβ oligomers from biological fluids, similar to the PMCA technique 

successfully developed by Claudio Soto for prion diseases. The same approach will be used to amplify and 

detect misfolded tau. 

• Biobanking and neuropathological and molecular characterization of patients

We will collect systematically plasma, DNA and CSF from patients with different types of dementia and 

non-demented individuals, and perform autopsies for neuropathological characterization and biochemical 

and molecular studies. 


• Recessive A673V APP mutation: molecular mechanisms and development of a new therapeutic strategy 

for sporadic AD 

We have recently identified an APP mutation (A673V) that causes early-onset AD only in the homozygous 

state while the heterozygous carriers are not affected. This mutation strongly boosts the production and 

amyloidogenic properties of Aβ. However, the interaction of A673V-mutated and wild-type peptides 

inhibits amyloidogenesis and A�-mediated neurotoxicity (Di Fede et al., Science 2009, 323:1473-7). These 

findings are consistent with the observation that the A673V heterozygous carriers do not develop disease 

and offer grounds for a novel therapeutic strategy based on modified Aβ peptides. A research priority of 

our lab is to unravel the molecular mechanisms of the opposite effects of the A673V APP mutation in 

homo- or heterozygous state on amyloidogenesis and to develop a lead compound for AD therapy based 

on A673V-modified Aβ peptides or peptido-mimetic molecules. To accomplish these objectives we have 

generated a panel of transfected cells and transgenic C. elegans expressing human APP or Aβ with the 

A673V mutation, respectively, and transgenic mouse lines expressing A673V-mutated APP in the 

homozygous or heterozygous state. Furthermore, we have identified a prototypic lead compound 

corresponding to a six-mer Aβ peptide with the A673V substitution and are currently working on brain 

delivery systems. 

• Pathways leading to tau pathology 

A key event in AD pathogenesis is the hyperphosphorylation, misfolding and aggregation of the 

microtubule-associated protein tau leading to formation of neurofibrillary tangles, disruption of the 

neuronal cytoskeleton and neurodegeneration. Following the “Aβ cascade hypothesis” this event is 

triggered by aggregated Aβ species, particularly oligomeric assembly intermediates. However, the 

pathways linking Aβ and tau pathology are unknown. This is largely due to lack of animal models able to 

reproduce these two central aspects of AD. We have developed a mouse model of prion disease that 

shows a secondary tauopathy following deposition of PrP amyloid (Giaccone et al., Neurobiol. Aging 2008, 

29:1864-73). This model will be used to investigate the molecular basis of tau pathology induced by 

deposition of an amyloid protein. 

• Role of nuclear tau in neurodegeneration in fronto-temporal dementia 

Tau is the major microtubule-associated protein of neurons and its primary role is to promote assembly 

and stabilization of microtubules required for morphogenesis and axonal transport. We have recently 

found that tau plays an important role also in chromosome stability, and mutations in the tau gene cause 

chromosome aberrations in peripheral cells in addition to formation of neurofibrillary tangles in neurons 

and glial cells. Our objective is to elucidate the molecular mechanisms underlying the genomic instability 

due to tau mutations using cellular models, and to investigate the contribution of the aneuploidy caused 

by mutated tau to neurodegeneration using a transgenic mouse model of tauopathy. 


CLINICA/PRECLINICA 

- Genetic susceptibility to Alzheimer’s disease (AD) 


- Biomarkers 

- Developing competitive animal models to study AD 

- Studying early onset forms of AD to follow-up disease progression 

- Cinical trials with or without involvement of pharmaceutical companies 



- Standardization of diagnostic criteria 


- Translational research 


1. Titolo progetto Genoproteomics of Age Related Disorders (GuARD).

PI: Roberto Sitia Subproject leader: Fabrizio Tagliavini 

Ente finanziatore Cariplo Foundation 


Abstract del progetto Project 3.2: Neurodegeneration. The main goals of this project are: (1) 

biochemical analysis of misfolded protein aggregates from brains of 

patients with sporadic and genetic forms of Alzheimer disease and prion 

diseases; (2) proteome analysis on brain tissue from the same patients 

and from gender- and age-matched non-demented control individuals; (3) 

proteome analysis on brain tissue from a mouse model of variant 

Creutzfeldt-Jakob disease. 


Alzheimer's disease: development and validation of a multi-factorial 

protocol for the diagnosis and follow-up of disease in the prodromal and 

incipient phase. PI: Fabrizio Tagliavini 

Ente finanziatore Italian Ministry of Health 


Abstract del progetto This program has two primary aims: (1) the development and validation 

of a multi-factorial protocol that integrates molecular, imaging, 

neurophysiological, neuropsychological and behavioral data for early 

diagnosis of AD, in particular during the stage of MCI; (2) the validation of 

this diagnostic protocol within a public health network, and evaluation of 

the health-care, organization and economic implications of its transfer to 

the NHS. 


Molecular mechanisms underlying synaptic dysfunction in prototypic 

neurodegenerative diseases related to protein misfolding (nEUROsyn). PI: 


Ente finanziatore European Union/Italian Ministry of Health 


Abstract del progetto This project is designed to investigate the mechanisms that lead to 

synaptic impairment and demise of neurons in AD and Huntington 

diseases using cellular and animal models, and patient material. The main 

goals are to: (1) study synaptic dysfunction; (2) identify alterations of 

synaptic and dendritic spine remodelling; (3) monitor changes in receptor 

trafficking at the synapse and disturbances in trafficking of different 

cellular components. 


A randomized, double-blind pilot study versus placebo for the evaluation 

of the efficacy of doxycycline administered by oral route in patients 

affected by Creutzfeld-Jakob disease. PI: Fabrizio Tagliavini 

Ente finanziatore Italian Italian Agency of Drug (AIFA) 


Abstract del progetto This project is a multicentre phase II clinical trial to test the effectiveness 

of an anti-amyloidogenic molecule (doxycline) identified in experimental 

models. 


Biological markers of neurodegeneration to be used in clinical practice for 

early and differential diagnosis of degenerative dementias, the 

identification of fast and slow progressing forms, and the evaluation of 

response to treatment. PI: Gianfranco Spalletta, Project Leader: Fabrizio 

Tagliavini 

Ente finanziatore Italian Ministry of Health 


Abstract del progetto The main goal of the project is the identification of peripheral markers of 

neurodegeneration in Alzheimer disease, Levy body dementia,

frontotemporal dementia and prion diseases to allow (1) early differential 

diagnosis, (2) identification of fast and slow progressing forms, (3) 

evaluation of response to therapy.


Contatti 

Tel. 

E mail 

Via Ardeatina, 306 – 00179 Rome 

06-51501575 





Area di interesse identificata Mild Cognitive Impairment 






Behavioral and Psychological Predictors of Cognitive Outcome in Subjects 

with Mild Cognitive Impairment: the Role of Non-conventional 

Neuroimaging and Serotonergic Genes 


01/09/2010 – 31/08/2012 

The present project will identify complex behavioural-endophenotypes, 

of behavioral-neuroimaging-genetics nature, useful for predicting the 

cognitive course of neurodegenerative dementias starting from their 

preclinical phase or MCI. In particular results will help to predict the 

outcome of the cognitive impairment, the relationship between cortical 

and subcortical brain atrophy and neuropsychiatric-neuropsychological 

longitudinal clinical manifestations, and a composite neuroimagingserotonin 

genetics index for better prediction of behavioral expression 

during the 18 months longitudinal course of the project. In order to do 

this an effort will be made to try to make the predictive capability of each 

clinical marker reliable for every single patient. 

Two-hundred people with a diagnosis of MCI will be recruited in this 

study. These patients will be drug free from psychopharmacological and 

acetilcholinesterase inhibitor drugs. One-hundred healthy controls (HC) 

will be also recruited. The subjects included will carry out an in depth 

anamnesis evaluation, a diagnostic evaluation for depression, psychosis, 

and apathy associated with dementia, and a psycho-behavioural 

dimensional assessment using a battery of tests including: 

Neuropsychiatric Inventory; CERAD Disforia; Dementia Apathy Interview 

Rating, Apathy Scale and Pittsburgh Sleep Quality Index. Moreover, the 

subjects will be evaluated through the MMSE, the Mental Deterioration 

Battery (MDB) and tests for basic and instrumental functional activities. 

This is a mixed case-control study (MCI at the baseline vs. HC) and 

longitudinal study (18-month follow-up only for MCI subjects). The first 

six months of research will be used for the inclusion of HC and MCI 

subjects in the study. Each cognitive and behavioural battery tests and 

diagnostic criteria will be administered in the MCI subjects every six 

months until 18 months from the first evaluation. Caregiver stress will be 

monitored at the baseline and the end of the follow-up. Image acquisition 

will be done at the baseline: all subjects will be examined using a 3 Tesla

Allegra MR Imager with a standard quadrature head coil. Participants will 

undergo the same MR imaging protocol including whole-brain T2*weighted, 

T1-weighted and DTI scanning. Standard clinical sequences 

(FLAIR and T2-weighted images) will be acquired on the same subjects in 

order to exclude different pathologies. Image processing will be 

performed using FreeSurfer, FSL 4.1 and in-house developed software in 

Matlab vers. 6.5. The FreeSurfer toolkit will be employed to perform 

automatic cortical thickness reconstruction. The high resolution 

anatomical T1-weighted images will be also used to extract several region 

of interest corresponding to subcortical grey matter nuclei. The automatic 

segmentation of these nuclei will be obtained with the software FIRST, 

part of the FSL toolkit. Moreover, the DTI data will be coregistered to the 

T1 images in order to extract regional values of diffusion parameters (MD, 

FA) known to be indicative of tissue microstructure disruption, from the 

same nuclei identified in the previous step. The DTI data will be also 

employed to identify the main fiber tracts within the white matter. These 

tracts that will represent additional regions of interest from where 

extracting diffusion parameters. Genomic DNA will be purified from 200 

microl of human whole blood. Polymorphisms and haplotypes of several 

genes will be analyzed in this study: the Apolipoprotein epsilon 4 (ApoE4) 

variant will be investigated in HC and MCI subjects in order to verify its 

influence on behavioural manifestations and symptom progression; the 

102T/C polymorphisms of the 5HT2A receptor gene will be investigated 

for psychosis behavioural phenotype; to test the hypothesis that allelic 

variation in 5HTT gene-linked polymorphic region (5-HTTLPR) genotype is 

associated with behavioural syndromes and symptoms and cognitive 

outcome, a common 44-base pair deletion (s allele) polymorphism in the 

5-HTTLPR which associated with reduced 5HTT transcription efficiency 

and 5HT uptake in vitro will be studied. 

The main focus of the research in the field of complex disorders, such as 

dementias, is to found new complex predictors which are valid and 

reliable in the prediction of clinical outcome starting from the early 

preclinical phases. In particular, mixed clinical (behavioural) and biological 

(neuroimaging, genetics) predictors may help to precociously identify 

those subjects who have a rapid decline of cognitive performances and 

those who will respond positively to the treatment with a stable course of 

the cognitive performances and who also improve. Another interesting 

possibility is that exist behavioural variables which may predict the course 

of the illness in the case they will not improve at the end of the follow-up. 

The longitudinal branch of this project is devoted to identify if this 

possibility exists and which are the behavioural predictors in terms of 

frequency of behavioural disorders at the end of the follow-ups.


E. Diagnosis: neuropsychology

Nome Maurizio Balestrino 

Contatti 

010-3537078/7085/7066; mbalestrino@neurologia.unige.it; FAX 010- 

3538631; mobile phone 348-7814154 

Istituto/Dipartimento Department of Neuroscience, Ophtalmology and Genetics, University of 

Genova, Italy 


Dementia of Alzheimer's type represents about 40% of the cases where a last will is challenged based on 

real or supposed mental incapacity of the testator (1). Usually, elder people find in their last will the 

reassurance that after their death their loved ones will have the resources to carry on their lives in 

agreement with the old person’s desires. However, when a person suffers (or is suspected to suffer) from 

Alzheimer’s disease, their last will is often disputed, on the ground that he/she may not have been 

mentally competent to make decisions. Thus, the peace of mind that comes with having written a 

thoughtful last will is considerably decreased. This is a growing problem, that is already painful from the 

social point of view (see for example http://elder-abuse-cyberray.blogspot.com/2007/06/where-there-iswill.html). 

Moreover, somebody may really take advantage of the declining cognitive status of an 

Alzheimer’s patient to have him/her write a will in his/her favor (2). The dispossessed heirs then claim that 

the will is invalid because the deceased was, at the time of writing it, incapacitated due to mental 

deterioration (3). 

In such cases, a judiciary trial follows, however the judgement is often difficult, because the writer of the 

will can no longer be examined, and the judgement must rely solely on the retrospective evaluation of 

medical records and of witnesses’ statements. Since both types of evidence are often incomplete or even 

discordant, the expert’s judgement is often difficult and flawed by obvious limitations. However, the last 

will has often been handwritten. If so, the quality of the handwriting may add very important elements to 

the expert’s evaluation, because it shifts the investigation from the retrospective medical records or 

witnesses’ claims to the direct observation of the deceased subject’s performance. The act of writing is by 

no means a mere movement of the hand and fingers, rather it is a cognitive performance from several 

viewpoints. The writer must (1) correctly build in his/her mind a suitable sentence, (2) translate words into 

graphic symbols, and (3) correctly arrange the writing in rows and columns, in such a way that it can be 

decoded. The first two tasks relate to speech abilities, and are usually negatively affected by aphasia. The 

third task implies a correct visuospatial ability. Both aphasia and visuospatial disturbances are prominent 

symptoms of Alzheimer’s type dementia (4). Accordingly, it has been shown that handwriting is 

compromised in demented patients (5,6,7,8). Even more important, a correlation exists between severity 

of handwriting disruption and severity of cognitive deficiency (for example, see Table 3 of reference (7) 

and Table 7 of reference (9)). 

While a wrong or confused handwriting is an obvious sign of mental impairment, the evaluation of this 

item in forensic psychiatry is currently hampered by the lack of the objective and quantitative tools that 

are required for a forensic examination. Although it has been demonstrated by several authors that 

writing is altered in demented patients (7,5,8,6), these data are difficult to apply to the evaluation of a 

single case, as it is required for judiciary purposes. By contrast, a quantitative score of handwriting that 

was able to identify severe mental deterioration with a high probability level would be an invaluable tool 

in the forensic evaluation. In preliminary research we addressed this issue by designing a semiquantitative 

standardized score system to evaluate handwriting, and investigated both its inter-rater reliability and its 

correlation with standard measures of cognitive performance (10). In that preliminary research we showed 

(a) that our semiquantitative handwriting scoring method has good interrater agreement and (b) that 

there is a statistically significant correlation between the degree of cognitive deterioration (as gauged by 

the score of both Mini Mental State Examination – MMSE - and of Milan Overall Dementia Assessment – 

MODA scale) and the handwriting score (10). 

We now propose to continue that research by testing a larger number of patients, to more definitely

quantify the correlation between severity of dementia and handwriting score. Specifically, we plan to (1) 

Repeat our investigation on a larger number of patients to hopefully better validate it. (2) Administer to 

our patients not only the MMSE and MODA tests, but also the Alzheimer´s disease assessment scale ( 

ADAS-COG), an internationally accepted tool for the assessment of Alzheimer’s disease. (3) Validate the 

test by identifying cutoff values of our handwriting score that reliably identify persons with normal mental 

capacity and persons with mild, moderate or severe dementia. 

We will test the hypothesis that (a) a writing score lower than a certain value (to be indentified in ou 

proposed research) reliably identifies patients whose mental deterioration is minimal or inexistent and (b) 

a writing score higher than a certain value (writing score=5 according to our preliminary research, see ref. 

10) reliably identifies persons with severe mental deterioration. 

We will carry out our research using a protocol similar to our published work (10). We will exclude patients 

having dementia not of Alzheimer's type, patients having severe motor deficit in the dominant hand, and 

patients having aphasia so severe as to prevent communication altogether. 

We believe that our reseach will help to bring justice and peace of mind to Alzheimer's disease victims and 

to their families, by providing a method, scientifically rooted in clinical neuropsychology, that will help 

preventing abuse and exploitation of Alzheimer’s disease patients through their last will. While this is in 

itself an important goal of our research, we also believe that the quantification of the relationship 

between Alzheimer’s disease and handwriting will open a new way of testing cognitive impairment in 

Alzheimer’s disease, with important fallout to the entire field of neuropsychological testing of Alzheimer’s 

disease patients. 

References 

8) Shulman, K. I., Cohen, C. A., and Hull, I., (2005) Psychiatric issues in retrospective challenges of 

testamentary capacity. Int.J Geriatr.Psychiatry 20:63-69. 

9) Peer, I. N., (1981) Wills, testamentary capacity and undue influence. Bull.Am Acad Psychiatry Law 

9:15-22. 

10) Redmond, F. C., (1987) Testamentary capacity. Bull.Am Acad Psychiatry Law 15:247-256. 

11) Victor, M. and Ropper, A. H., (2005) Adams and Victor's Principles of Neurology. 8th Ed.: 

12) Werner, Perla, Rosenblum, Sara, Bar-On, Gady, Heinik, Jeremia, and Korczyn, Amos, (1-7-2006) 

Handwriting Process Variables Discriminating Mild Alzheimer's Disease and Mild Cognitive 

Impairment. Journals of Gerontology Series B: Psychological Sciences and Social Sciences 61:228- 

236. 

13) Croisile, B., (2005) [Writing, aging and Alzheimer's disease]. Psychol Neuropsychiatr.Vieil. 3:183- 

197. 

14) Silveri, Maria Caterina, Corda, Francesca, and Di Nardo, Miriam, (2007) Central and peripheral 

aspects of writing disorders in Alzheimer's disease. J Clin Exp Neuropsychol. 29:179-186. 

15) Forbes, Katrina E., Shanks, Michael F., and Venneri, Annalena, (1-3-2004) The evolution of 

dysgraphia in Alzheimer's disease. Brain Research Bulletin 63:19-24. 

16) Luzzatti, Claudio, Laiacona, Marcella, and Agazzi, Daniela, (2003) Multiple patterns of writing 

disorders in dementia of the Alzheimer type and their evolution. Neuropsychologia 41:759-772. 

17) Fontana P, Dagnino F, Cocito L, Balestrino M. (2008) Handwriting as a gauge of cognitive status: a 

novel forensic tool for posthumous evaluation of testamentary capacity. Neurol Sci. 29(4):257-61. 

18) Brazzelli, M., Capitani, E., Della Sala, S., Spinnler, H., and Zuffi, M., (1994) A neuropsychological 

instrument adding to the description of patients with suspected cortical dementia: the Milan 

overall dementia assessment. J Neurol Neurosurg.Psychiatry 57:1510-1517. 

19) Folstein, M. F., Robins, L. N., and Helzer, J. E., (1983) The Mini-Mental State Examination. Archives 

of General Psychiatry 40:812. 

Area di interesse identificata - Standardization of diagnostic criteria and diagnostic


Titolo progetto // 



// 

// 


- Multidisciplinary projects

Responsabile: Angiola Maria Fasanaro (Neurologo) 

Collaboratori: Raffaele Rea (Volontario Neuropsicologo) 

Anna Carotenuto ( Volontario Neuropsicologo) 

Maila D’Anonio (Volontario Neuropsicologo) 

Luisa Colucci ( Tirocinante Neuropsicologo) 

Sabrina Carpi ( Tirocinante Neuropsicologo) 

Massimiliano Bosco ( Tirocinante Neuropsicologo) 

Ivana Molino ( Tirocinante Neuropsicologo) 

Gabriella Finizio (Tirocinante Neuropsicologo) 

Antonio Ziello ( Tirocinante Neuropsicologo) 

La Unità Valutativa Alzheimer, da me diretta è attiva dal 2000, e fa parte della Struttura Semplice di 

Malattie Involutive Cerebrali, Dipartimento di Neuroscienze, AORN "A. Cardarelli". 

Le attività dell’ ambulatorio sono prevalentemente svolte su soggetti geriatrici. 

L’esperienza maturata nel corso di 10 anni di attività ha permesso di valutare oltre 900 pazienti , ed 

attualmente circa 400 sono in terapia regolare. I dati sono raccolti in cartelle cliniche ad hoc ed archiviati 

elettronicamente. 

I protocolli seguono le Linee Guida Internazionali e si avvalgano dell’ applicazione di test neuropsicologici 

specifici di approfondimento. Particolare attenzione viene dedicata all’ amnesic/non amnesic Mild Cognitive 

Impairment e ai fattori vascolari associati alle demenze. 

I pazienti sono controllati a distanza di uno, tre e sei mesi dall’inizio della terapia, ed ogni sei mesi 

successivamente. 

Oltre alle attività diagnostiche e di monitoraggio l’Unità svolge un servizio di counseling diretto ai caregiver 

e programmi di riabilitazione diretti a preservare capacità residue. 

Attualmente il centro si avvale della collaborazione di 9 neuropsicologi tra volontari e tirocinanti che 

costantemente si occupano delle attività ambulatoriali e di ricerca. In quest’ ambito è stata svolta una 

collaborazione regolare nel triennio 2005-2007 con L’ Università di Tunisi per un progetto di 

implementazione delle metodiche neuropsicologiche.(progetto finanziato dal Ministero degli Affari Esteri.) 

Abbiamo una collaborazione stabile con la Cattedra di Neuropsicologia Clinica della Seconda Università 

degli Studi di Napoli. e con il Dipartimento di Medicina Sperimentale e Sanità Pubblica dell’Università di 

Camerino. E in corso di pubblicazione una ricerca realizzata insieme all’Università Orientale di Napoli e all’ 

Università La Sapienza di Roma.

I principali progetti di ricerca attualmente attivi 

D. 

1) Caratteristiche neuropsicologiche differenziali tra pazienti con Demenza fronto-temporale e con 

variante frontale della Malattia di Alzheimer. 

2) Il contributo del network frontale alla genesi delle allucinazioni in pazienti con malattia di Alzheimer e 

con malattia di Parkinson. 

3) I fenomeni attrattivi spaziali ed il closing-in 

4) Caratteristiche fMRI del Mild Cognitive Impairment Amnesico. 

5) Caratteristiche neuropsicologiche delle varianti atipiche della Malattia di Alzheimer. 

6) Valutazione cognitiva e monitoraggio degli effetti terapeutici dei trattamenti farmacologici 

(Rivastigmina patch, Precursore Colinergico Colina alfoscerato, RO5313534 ) sui sintomi cognitivi e 

comportamentali della malattia di Alzheimer e nelle forme con danno vascolare associato. 

7) Analisi dei costi sociali ed economici delle demenze. 

8) Arte visiva come proposta di riabilitazione cognitiva e motivazionale 

9) Depressione e Malattia di Alzheimer studio sui correlati gene-ambiente e risposta al trattamento 

farmacologico.

Nome Roberto Gallassi 

Contatti 

roberto.gallassi@unibo.it 

Istituto/Dipartimento Department of Neurological Sciences, University of Bologna 


Area di interesse identificata Earli diagnosis 



1) Centre for the Neurological Study of Brain Aging – Gallassi 

Roberto 

2) Neurogenetic Unit – Carelli Valerio 

3) Neuropathology Unit – Parchi Piero 

Department of Internal Medicine, Aging and Nephrology, University 

of Bologna 

1) MR Spectroscopy Unit – Lodi Raffaele 

Predictors of conversion from MCI to dementia 

Ente finanziatore Fondazione Gino Galletti 



Background 

MCI is a well known clinical condition intermediate between normal 

ageing and dementia. It is not completely ascertained if this entity is 

a beginning of a dementia or a clinical syndrome which can have 

different outcomes. Anyway, the annual conversion rate of MCI to 

dementia is variable in different previous studies, ranging from 

about 4% to 12% or more. The investigated factors of conversion 

taken into account are clinical aspects, neuropsychological tasks, 

conventional and functional neuroimaging measures, biological 

markers and genetic factors. 

Previous neuropsychological studies indicates that MCI may be 

characterized by subtle impairment in learning and delayed recall,

particularly in tasks of episodic memory and visuospatial memory, in 

executive functions, phonemic and semantic verbal fluency, etc. In 

our recent study, we found that a behavioural memory battery, 

visual attention, long-term verbal memory tasks and 

Neuropsychiatric Inventory caregiver distress and apathy scores, 

resulted in independent predictors of conversion to dementia. We 

also found that single domain impairment, including memory, has a 

minor probability to develop dementia than multiple domain 

impairment. 

Rationale 

We hypothesize that some cognitive functions and 

neuropsychological tasks are more sensitive in predicting the 

conversion of dementia in relationship with modifications in other 

clinical, biochemical, genetic and neuroimaging findings. 

Aim 

The aims of this project are: 

- to evaluate at baseline demographic features, clinical 

characteristics, cognitive and behavioural profile, biochemical 

and genetic findings in a sample of MCI patients of amnesic or 

not amnesic type and with single or multiple domain 

impairment. 

- to perform clinical and MR neuroimaging longitudinal 

observation during three years of the MCI sample, evaluating 

the evolution of the single patients, in particular the conversion 

to dementia 

- to correlate with a multi-dimensional approach the relationship 

between clinical, cognitive and behavioural findings and the 

other methods of investigation such as quantitative MR 

neuroimaging measures, haematic and cerebrospinal fluid 

biological data and the genetic profile of patients 

- to evaluated the independent predictors of conversion from 

MCI to the various type of dementia

F. Biobanking Bioinformatics


G. Ambient assisted living and socioeconomics

Roberto Monastero, MD, PhD 

Lab of Epidemiology and Psychology of Aging and Dementia 

Dept of Clinical Neuroscience 

University of Palermo 

Via La Loggia 1 

90129 - Palermo, Italy 

Tel. +39-091-6555185 

Fax. +39-091-6555113 

email. roberto.monastero@ki.se 

roberto.monastero@unipa.it 

PROGETTO2 

Type: Ambient Assisted Living (AAL) Joint Program, 2008 

Title: Home-based Empowered Living for Parkinson’s disease Patients (HELP) 

Responsibles: Prof. Giuseppina Campisi, Dr. Roberto Monastero 

Amount of fund: ......................... 

Lenght: 3-year project 

Abstract 

Medical and social development have produced remarkable changes in the population distribution at 

worldwide level, leading to a dramatical increase in the life expectancy of the human population. the higher 

life expectancy is usually associated with the appearance of chronic conditions, such as Parkinson's disease 

(PD), and associated comorbidites, which burdens and have a great impact in the quality of life of both the 

persons suffering the illness and his/her relatives and caregivers. Besides, chronic conditions also derive in 

higher costs for the care delivery process, becoming a problem for the whole society. In this sense, the 

technological advances of the last decades can, and should, provide solutions to better manage the 

treatment for the patients, reducing the social and economic impact of the chronic conditions. The Homebased 

Empowered Living for Parkinson’s disease Patients project will include a system able to administer 

drug therapy in a controlled and either continuous or on-demand basis, to control disease progression and 

to mitigate PD symptoms. Accordingly, it will give a comprehensive service to help Parkinson's patients to 

better cope with their illness and reduce comorbidity.

Country: ITALY 

Contact person: Alessandra Pedrocchi (Politecnico di Milano) 

Date: 14th Jan 2010 


TOPIC 1 

Neuro-rehabilitation and brain functional imaging: rehabilitation customization driven by best 

exploitation of cerebral plasticity and health system sustainability 

The spreading of neuro-motor pathologies has a great impact on the healthcare system European 

Countries, being among the first causes of death (stroke is the third) but utmost being the first case of high 

severity long-term disabilities. The recovery and the progress of neuromotor diseases is strongly affected 

by the pathogenesis but also by the rehabilitation training, which has the purpose to best exploit the 

cerebral plasticity governing neural motor control reorganization. 

Beside the traditional therapist assisted training, different novel solutions has spread in the latest years 

based on robotic systems, functional electrical stimulation (neuroprostheses) and hybrid systems where 

robots and electrical stimulation are integrated. 

Nonetheless, the international scientific community still lack of a complete well supported comparative 

validation of these approaches and consequently a specific customization of the best training cocktail for 

each patient is still not documented by scientific results and it is mainly devoted to clinician personal 

convictions and experience. 

Especially, we claim that in term of motor recovery and motor relearning, i.e. cerebral re-organization, 

there is no clear scientific substrate to support the use of robotic devices and/or neuroprostheses aside or 

in alternative to standard rehabilitation. 

However, in the latest decade the possibility to investigate in-vivo cerebral activations associated to 

movement execution has become accessible , thanks to fMRI, EEG and fNIRS,… 

The challenge to be tackle in the next future is to integrate these two aspects aiming at studying the 

impact of rehabilitation selections on the cerebral plasticity reorganization in neuromotor pathologies. 

Beside the efficacy of those methods and their best exploitation, it also needs to be tested the 

sustainability of these solutions for the long term economical, environmental and social impact of 

biomedical technologies in the rehabilitation of neuromotor pathologies.

Nome Alberto Pilotto 

Contatti 

Phone: +39 0882 410271 

Fax: +39 0882 410271 

E-mail: alberto.pilotto@operapadrepio.it 

Istituto/Dipartimento Unità Operativa di Geriatria & Laboratorio di Gerontologia-Geriatria 

Dipartimento di Scienze Mediche 

IRCCS Casa Sollievo della Sofferenza 

Viale Cappuccini, 1 – 71013 San Giovanni Rotondo (FG) 


Sporadic Alzheimer’s disease (AD) is a progressive neurodegenerative disorder occurring predominantly in 

older age. The prevalence of AD rise from 20% after 75 years to 30% after 85 years, and about 5% of 

people aged 65 years or older have AD. With about 3 to 4 million people affected in the United States and 

about 350,000 new cases per year, AD is the most frequent cause of dementia in U.S. and in Western 

countries. Among the main cause leading to AD, the deficit of cholinergic system plays a major role. 

Accordingly, one of the most common therapy for the symptomatic treatment of AD is the block of 

acetylcholinesterase (AChE) by means of acetylcholinesterase inhibitors. The inhibition of AChE increases 

the concentration of acetylcholine (ACh) in the synaptic cleft, thus restoring the physiological effects of 

ACh within the central nervous system. Recent studies reported a significant benefits of 

acetylcholinesterase inhibitors vs placebo on cognitive function, activities of daily living, and behavior. 

These improvements, however, are not always detectable in clinical practice. Thus, it has been recently 

suggested that the detection of improvements on cognitive function, activities of daily living, and behavior 

may be obtained by the concomitant use of drugs acting on other pathogenetic AD mechanism. In 

particular the European Medical Agency (EMEA) recommend the use of the AChE inhibitors donepezil and 

rivastigmine in mild-to-moderate AD, and the addition of memantine in moderate-to-severe AD. 

Memantine block the glutamate receptor, limiting the uncontrolled entrance of Ca 2+ ion in the 

postsynaptic neuron, thus delaying the Ca 2+ -excess neurodegeneration. Most studies reported that 

interindividual differences in response to these drugs may be due to variability in drug metabolism related 

to behavioral, clinical, and genetic factors, mainly hereditary polymorphisms of drug-metabolizing and 

drug-transporting enzymes. Final objective of this proposal is to identify the genetic component 

underlying the response/non response to the most common drugs currently used in the treatment of AD, 

i.e. donepezil, rivastigmine and memantine, throughout the following specific objectives: 1) identification 

of significant difference in the distribution of the genotypes of the CYP2D6 gene polymorphisms among 

patients responders/non-responders to donepezil treatment; 2) identification of the specific DNA 

alteration producing modifications in the CYP2D6 enzyme activity; 3) confirmation of the modification of 

the enzyme activity by means of donepezil dosage in vivo; 4) identification of significant difference in the 

distribution of the genotypes of the acetylcholinesterase gene polymorphisms among patients 

responders/non-responders to rivastigmine treatment; 5) identification of the specific DNA alteration 

producing modifications in the acetylcholinesterase-mediated hydrolysis of rivastigmine; 6) confirmation 

of the modification of the enzyme activity by means of rivastigmine dosage in vivo; 7) identification of 

significant difference in the distribution of the genotypes of the OCT2 gene polymorphisms among 

patients responders/non-responders to memantine treatment; 8) identification of the specific DNA 

alteration producing modifications in the OCT2 transporter of memantine; 9) confirmation of the 

modification of the enzyme activity by means of memantine dosage in vivo. We expected that the 

identification of functional polymorphisms in the CYP2D6, AChE and OCT2 genes may influence the clinical 

efficacy of donepezil, rivastigmine and memantine in AD patients, and may be useful in identifying 

subgroups of AD patients with different clinical response to treatment. Criteria and indicators to verify

esults are: 1) the number of patients enrolled in the study in the first 6 months; 2) the number of 

genotypes investigated at the end of the 1 th year of the project; 3) the number of specific gene alteration 

influencing the enzyme activity at the 2 nd year of the project; 4) the quantitative data obtained from 

plasma drug dosage at the end of the 2 nd year of the project. This project is coherent with the ministerial 

guidelines for the I.R.C.C.S. and well integrate the clinical practice in neurology. 







Pharmacogenetics of drugs for Alzheimer’s Disease 

Effect of weight loss on metabolic, functional, cognitive status and 

biological markers of longevity in obese frail elderly 


Two years 

Obesity, and in particular abdominal obesity causes serious metabolic and 

medical complications and impairs quality of life. Moreover, in elderly 

persons, obesity can lead to frailty by promoting chronic inflammation 

and by exacerbating the decline in strength, endurance, balance and 

mobility associated with aging and physical inactivity. Weight loss, 

induced by calorie restriction and/or physical exercise, simultaneously 

improves multiple metabolic risk factors for cardiovascular disease and 

other medical abnormalities associated with obesity, and reduce 

morbidity and mortality. However, the appropriate treatment for obesity 

in elderly persons is controversial because of the potential harmful 

effects of weight loss on bone and muscle mass. It has been reported that 

weight loss can also modify biological pathways at the cellular level. 

Insulin and of insulin-like growth factor-1 (IGF-1) have been involved in 

cellular caloric equilibrium and other mechanisms, i.e. oxidative stress 

and inflammation. Thus genetic polymorphisms in genes involved in these 

metabolic pathways such as apolipoprotein E (APOE), sterol regulatory 

element-binding protein cleavage-activating protein (SCAP), peroxisome 

proliferators-activated receptor gamma-2 (PPR�), �-2B-adrenergic 

receptor (ADRA2B), acyl-CoA synthetase 5 (ACSL5) and interleukin-6 (IL-6) 

may influence the effect of weight loss on biological mechanisms and 

possibly on functional and cognitive status in the frail elderly. It is possible 

that weight loss in obese elderly persons can be beneficial by improving 

metabolic, functional and cognitive status, or harmful by causing a 

decrease in muscle and bone mass. However, the clinical and 

physiological effects of weight loss in obese elderly subjects have never 

been carefully evaluated. Thus, appropriate treatment for this rapidly 

increasing segment of the elderly population remains controversial. The 

purpose of this project is to advance our scientific knowledge by 

determining the effects of weight loss on metabolic, functional and 

cognitive status in obese elderly subjects. Moreover, we will evaluate the 

role of weight loss on biological markers of metabolic health and 

longevity, i.e. serum level of Insulin, insulin-like growth factor-1 (IGF-1), 

transforming growth factor-� (TGF-�), tumor-necrosis factor-� (TNF-�), 

interleukin-1 (IL-1) and C-Reactive Protein (CRP), and their relationships 

with genetic polymorphisms in APOE, SCAP, PPR-�, ADRA2B, ACSL5 and





IL-6 genes. As biomarker of longevity we will also evaluate the 

accumulation of mitochondrial (mt) DNA mutation through the analysis of 

the D310 mononucleotide repeat of mtDNA. 

Smart Home for Elderly People (HOPE) 

Ministero della salute/Ministero dell’Università e della Ricerca Scientifica 

Two years 

Since Europe’s and worldwide population grows older, the need for care 

is growing as well as people spend more money to get it. The solution 

Smart Home for Elderly People (HOPE) promises to reduce the need for 

carers, improve the life of older people and cut the cost of assistance. The 

main objective of the “Hope” project is to produce an Integrated 

Computer Technology (ICT) solution that will help the elderly people, 

specifically those that suffer with the Alzheimer s disease (AD), achieve a 

richer and more independent lifestyle. Dementia causes long and 

oppressive suffering to patients and their relatives and imposes 

enormous costs on society. About 25 million people suffered from 

dementia in recent years. As a 4-fold increase of this number is expected 

by 2050, dementia is one main health issue of the next decades. AD 

covers 50-70% of all dementia cases, no cure exists, and effective and 

reliable early diagnostic techniques are lacking. Early diagnosis and 

progress monitoring of AD is a central part of treatment until future drugs 

and prevention strategies become available. Elderly people with AD 

spend most of their time at home. The “HOPE” solution consists of an 

integrated, smart platform that will enable the elderly people with AD to 

use innovative technology for a more independent life, easy access to 

information, monitor their health, and serve as a source of inspiration for 

users as well as for people working with assistive devices. Moreover, it 

will enable them to perform by themselves activities they were not able 

to do before and which are important for their daily personal life. The 

main advantage of the proposed system is that it provides a basis for 

integrating services for the elderly population while they are at home. 

HOPE is a budgeted solution that will be installed at the elderly people’s 

homes, and will provide services for (a) life-long, self organized, 

appropriate educational environment and access to information, (b) care 

management and health support, (c) self monitoring and decision making. 

Optionally, the user will be able to activate software which automatically 

establishes the necessary interactive, triple-play connection for receiving 

tele-help and tele-assistance from specialized service providers, doctors 

or other medical personnel. The proposed application will be intelligent 

enough to offer safety in terms of controlling efficiently the home 

environment, economy in terms of controlling and decreasing the need 

for external help, and convenience in terms of adjusting the operation of 

connected sub-systems to achieve the best possible user experience.

Nome Vincenzo Mario Bruno GIORGINO 

Contatti 



TEL. (WK) (0039) (0)11 6706093 e-mail vincenzo.giorgino@unito.it 

Cell. 3280380926 

Department of Social Sciences - Faculty of Economics - University of 

Torino 

A transdisciplinary oriented proposal from a social sciences perspective 

From the bedside to the bench and return 

The major challenge in the field of senile dementia is represented by the existent gap between different 

disciplinary approaches, namely between bio-medical and social/human sciences. The project aims to 

answer to this gap within a transdisciplinary perspective, mainly based on the effort to integrate 

experiential data coming from social sciences methods with biomedical ones. The former are considered a 

possible source of knowledge that, under certain conditions, can be meshed with the latter. 

The research strategy is based on the recruitment of around 50 patients with mild cognitive 

impairment or symptoms of early Alzheimer’s disease and their caregivers, both formal and informal. 

Selection criteria of participants within a grounded theory methodology will be based on the greater 

similarity, i.e. the most similar personal/social characteristics as well as of the context. 

The research design is a mixed method approach oriented to collect data from the experiential 

level principally through methods such as journals, systematic observation, in-depth interviews and focus 

groups with the caregivers, both formal and informal. The same could be said for Alzheimer’s patients as 

far as possible and, in this regard, first-person observation is of the utmost importance as it allows a 

knowledge that is unavailable with usual third person methods; the former can be assessed with adequate 

criteria as recent literature is showing. 

One major assumption is that the loss of cognitive functions could not mean the loss of the 

awareness of this painful condition; moreover, if the patient is usually unable to express his/her feelings, it 

does not mean that he/she does not have feelings at all. So far, we can develop adequate tools of 

systematic observation to explore these more subtle processes. 

The project may include a two month period of an experimental training for caregivers in which 

they could learn to practice some contemplative methods in their everyday life. This is a goal of 

translational relevance that can include the patients, depending by their specific condition, and calling for 

a specific kind of exercises. Following this perspective, we can set up two groups of around 25 caregivers 

each. To the first group is offered a two month contemplative practice program, while the second will 

follow the current standard service provision. 

If in the literature social ties are recognized of major importance both at health promotion and 

prevention level than during the support for the sick, they do not exist in a vacuum: they call for personal

commitment, i.e. emotional, cognitive and economic resources. Social support and self healing are here 

intended as nurtured by a disciplined learned skills of contemplative techniques: see for example the 

literature about TM and mindfulness approaches which provide a cultural maintenance to our biological 

self. 

As a further development, a contribution by an economist could develop a specific research 

strategy for a study about the costs of this kind of intervention compared with standard ones. 

A valuable consequence of this approach could also be appreciated at cognitive level: a fresh 

understanding of difficult situations could emerge in everyday life by the caregivers or in the professional 

realm, suggesting new opportunities of managing the interactional lived situation and opening the way to 

new possible treatments. The hypothesis is that contemplative methods could enhance not only the 

coping function but also the general interactional process related to the management of this illness, 

including a better interaction and understanding with/of the patient’s experience, currently shadowed in 

clinical research. One can envisages also the consequences that this kind of approach could have at the 

governance level, in which caregivers and, when possible, patients can be co-producers of the healing 

process, as recent NHS policy programs show in the UK. A broader encompassing framework based on an 

experiential-empirical perspective could enhance the scientific development and open to more effective 

intervention. 


Vincenzo Mario Bruno Giorgino 

Experimental methods oriented to personal experience 

Transdisciplinary research 

Sociology of health, of health profession and informal care

Mapping of Italian research excellence in Neurodegenerative - Apre

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