GAMeets - the Instituto Gulbenkian de Ciência - Fundação Calouste ...

GULBENKIAN ALUMNI MEETINGS
(GAMeets)
6th Edition
15th Anniversary of the Cúria Meetings
Palace Hotel Cúria
27-28 December 2010
1

Welcome!
This year, we have made great efforts to bring you all to Curia.
Not only to reminisce of those “golden” days in the past of course,
but most specially to contribute to the continuation of our future. We
hope that these two days that we spend together will foster many
future collaborations, scientific and other, thus strengthening our
network as it proliferates through our new entrants, many for whom,
Curia this year is the very first time, and to whom we warmly
welcome and hope that this event and this singular location will be
enjoyed in just the same manner as it was in the past!
Monica Dias, Sofia Araujo and Greta Martins
Some important links for networking:
IGC Facebook
http://www.facebook.com/InstitutoGulbenkianCiencia
IGC PhD Alumni Facebook Group
http://www.facebook.com/home.php?sk=group_178917335470253
2

MONDAY 27 DECEMBER
Programme
Morning: Arrivals and check-in at the Curia Palace Hotel
12h30: Lunch and Opening Session
14h00: SESSION ONE
PGDBM2 - Cláudio Gomes, Margarida Trindade,
Guilherme Neves. PGDB2 - David Cristina, Helena
Soares, Eduardo Silva. PGDB4 - Joana Sá, Paulo
Ribeiro, Tiago Carvalho. PGDBM3 - Pedro Brites,
Perpetua Ó, José Leal. PIBS - Lilia Perfeito, Dinis
Calado, Cristina Borges. PGDBM4 - Alexandra
Capela, Carla Martins. PDBC - Isabel Duarte.
16h30: SESSION TWO: Coffee Break and Poster Session
Mónica Bettencourt-Dias, Sofia Araújo, Ana Teresa
Tavares, Pedro Domingos, Ofélia Carvalho, Francisco
Dionísio, Marta Nunes, Vera Teixeira, Rui Peixoto,
Daniel Sobral, Ulla-Maj Fiuza, Paula Duque, Greta
Martins.
18h00: SESSION THREE
PGDB1 - Vanessa Zuzarte, Miguel Gaspar, Luís
Soares. PGDB5 - Ana Mateus, Ricardo Silva, Catia
Proença. PGDB3 - César Evaristo, José Antão,
Catarina Homem. PGDBM5 - Bruno Silva-Santos, Rita
Nunes, Susana Nery, Isabel Gordo. PGDMB6 - Rita
Teodoro, Luis Teixeira. PGDBM7 - Tiago Carneiro, Mª
João Leão
20h30: Dinner
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22h00: Jonathan Howard,
TUESDAY 28 DECEMBER
University of Cologne, Germany and IGC SAB Member
“How to build a perfect institute”
9h30-10h40: SESSION FOUR
PGDBM1- António Jacinto, Luís Martins, José Belo,
Isabel Palmeirim, PGFMA-Ana Cerdeira, Sofia Braga,
INDP-Maria Inês Vicente, Pedro Ferreira.
10h45-11h00: Coffee break
11h00-12h00: Maria Leptin, Director, EMBO, Germany
“My Life in Science”
12h00-12h30: Closing Session:
António Coutinho, IGC and Diogo Lucena, FCG
13h00: Lunch – Leitão at Anadia Museum
4

ABSTRACTS FOR ORAL PRESENTATIONS
Monday 27 December
SESSION ONE
6

Protein deposition pathways at the synaptic milieu
Claudio M. Gomes
Instituto de Tecnologia Química e Biológica, Universidade Nova de
Lisboa, Oeiras
My laboratory investigates the biology and biophysics of protein
folding, the essential cellular process through which proteins acquire
a functional conformation. In recent years we have addressed how
this process is affected in neurodegenerative and metabolic diseases,
resulting in protein misfolding or aggregation (1-3). Protein
deposition as amyloid oligomers in the human nervous system is
characteristic of neurodegenerative diseases such as Alzheimer’s and
Parkinson’s. The identification of cellular modulators of protein
deposition remains a challenging issue, which has been the focus of
our recent attention.
In this brief talk I will overview our approach to understand the
biology and mechanisms of protein aggregation in the nervous
system. In particular I will focus on the role played by the unique
chemical biology of the synaptic environment, namely high
concentrations of metal ions and secreted proteins in a molecular and
physically confined environment. To illustrate this aspect I will
overview our recent progresses on the analysis of amyloid formation
by S100 proteins, which are upregulated in the brain in amyloid
diseases (Alzheimer’s and ALS), and the cross talk with the amyloidbeta
peptide under conditions mimicking the synaptic environment
(4).
1. Kim, S., Leal, S. S., Ben Halevy, D., Gomes, C. M., and Lev, S.
(2010) J Biol Chem 285(18), 13839-13849
2. Henriques, B. J., Rodrigues, J. V., Olsen, R. K., Bross, P., and
Gomes, C. M. (2009) J Biol Chem 284(7), 4222-4229
3. Correia, A. R., Adinolfi, S., Pastore, A., and Gomes, C. M. (2006)
Biochem J 398(3), 605-611
4. Fritz, G., Botelho, H. M., Morozova Roche, L., and Gomes, C. M.
(2010) FEBS J, in press
8

Science is funding
Margarida Trindade
Instituto de Medicina Molecular, Faculdade de Medicina da
Universidade de Lisboa
Margarida Trindade is currently responsible for Science Funding, at
the Communication and Training Unit at the Instituto de Medicina
Molecular (IMM) in Lisbon. Her current responsibilities include
supporting researchers assembling national and international
research funding proposals, grant application management, contract
negotiation and training on grant writing for graduate students. At
the interface of the Institute’s administrative services and the
researchers, Margarida’s goal is to foster an environment for
additional science funding at IMM.
9

TRANSGENIC MICE
Guilherme Neves (1) Mala Shah (2) Petros Liodis (1) and Vassilis
Pachnis (1)
(1) Molecular Neurobiology Division, NIMR, London, UK
(2) The School of Pharmacy, UCL, London, UK
Defects in inhibitory circuits in the cortex often underpin the emersion
of seizure activity. However the relative importance of somatic versus
dendritic inhibition in the pathogenesis of epilepsy remains unclear.
The LIM homeodomain transcription factor Lhx6 is essential for the
differentiation of the parvalbumin (Pva) and somatostatin (Sst)
expressing subpopulations of GABAergic interneurons, and for the
normal development of inhibitory circuits in the mouse cortex. Null
mutations in Lhx6 result in a pronounced deficit in synaptic inhibition
and anatomical signs of severe seizure activity. Interestingly,
hypomorphic Lhx6 mutants show a remarkably selective deficit in the
differentiation of Sst+ interneurons. Moreover, these animals show
behavioural, histological and electroencephalographic signs of
recurrent seizure activity, starting from early adulthood. Consistent
with a selective defect in the differentiation of Sst+ interneurons,
which project primarily to the dendritic compartment of principal
neurons, we have observed characteristic deficits in dendritic
inhibition but normal inhibitory input onto the somatic compartment
of pyramidal cells. These data implicate the Sst+ interneuron
population as an important control system that normally prevents the
emergence of seizure activity.
10

Technology Transfer in the Greater Lisbon Area
David Cristina, Bruno Reynolds, Lígia Martins and António Coutinho
IGC, ITQB
Technology Transfer (TT) is the exploitation, commercial or
otherwise, of scientific research for the direct benefit of society. It's
activities include sourcing invention disclosures, guaranteeing patent
protection, licensing technologies, forming spin-out companies and
managing consultancy opportunities for scientists. The quality of
Portuguese Life Sciences research has increased significantly over the
last 10 years, however the TT structures have not kept up with this
development. As part of our project, we are developing a TT Office
addressing this specific need for the Greater Lisbon area. We are
doing this by analyzing different international TT models and
adjusting them to the national context. We have already started
providing services to researchers and, so far, we have been able to
assist in the formation of a "start-up", Acellera Therapeutics.
11

Hierarchically regulated exocytosis controls signal amplification at the
immunological synapse and is targeted by HIV-1
Helena Martins Soares (1) Françoise Porrot (2) Olivier Schwartz (2)
Maria-Isabel Thoulouze (1) and Andrés Alcover (1)
(1) Lymphocyte Cell Biology Unit
(2) Virus an Immunity Unit. Institut Pasteur
T cell receptor (TCR) signaling is triggered and controlled at
immunological synapses. TCR and signaling effectors concentrate at
the synapse, form dynamic signaling clusters, being then differentially
sorted. This facilitates the induction, amplification and extinction of
TCR signaling that drives T cell activation. Intracellular vesicle
transport targets the TCR, the tyrosine kinase Lck and the adaptor
LAT to the synapse, but the regulation of this transport and its
significance for TCR signaling remain unknown. We show that HIV-1
dissects this vesicle transport unveiling that Lck, TCR and LAT are in
distinct vesicular compartments that concomitantly polarize to the
synapse. Lck compartment fuses first with the plasma membrane in a
calcium-independent manner, then regulating the calcium-dependent
fusion of TCR and LAT compartments, and the subsequent LAT
phosphorylation. Therefore, a hierarchically regulated exocytocitic
process drives signal amplification at the immunological synapse.
HIV-1 breaks this process by retaining Lck in endosomes therefore
uncoupling LAT from the TCR.
12

Material Systems for targeting and reversing ischemia
Eduardo A. Silva
Wyss Institute for Biologically Inspired Engineering - Harvard
University
Spatially and temporally regulated signaling between and within cell
populations and the extracellular matrix regulate tissue homeostasis,
pathology and regeneration, and polymeric materials that can mimic
or enhance this communication have the potential to intervene in
these processes in a therapeutic manner. These cell instructive
polymer systems provide insoluble signaling molecules and cues
(e.g., adhesion peptides) or soluble signaling molecules (e.g., growth
factors) alone or in specific combinations to either host tissue cells or
to transplanted cells to regulate their activation, multiplication and
differentiation. Multiple aspects of regeneration must be considered to
design effective approaches to enable functional tissue replacement,
and this issue has been addressed in the context of angiogenesis. The
systems described in this research could represents an attractive new
generation of therapeutic delivery vehicle for treatment of
cardiovascular diseases, as it combines long term in vivo therapeutic
benefit with minimally invasive delivery.
13

Dead mules: when strong post-zygotic isolation is not sufficient to
prevent mating
Joana Gonçalves-Sá (1,2,3) and Andrew Murray (1,2)
(1) MCB, Harvard University, Cambridge, MA 02138, USA
(2) FAS Center for Systems Biology, Harvard University, Cambridge,
MA 02138, USA
(3) Gulbenkian Ph.D. Program in Biomedicine, IGC, Oeiras, Portugal
Speciation, the paradigm has it, happens when two populations can
no longer interbreed and give rise to fertile progeny. Thus, the
prediction is that if two species share an ecological niche and their
hybrids have very strong fitness defects, there would be an
advantage in evolving mechanisms to prevent mating events between
them.
In Saccharomyces cerevisiae mating occurs when two haploid cells of
opposite mating types fuse. The two cells communicate through
secreted pheromones and the corresponding transmembrane
receptors and we asked whether, like in other species, receptor
specificity could be playing a role in reproductive isolation.
We looked at the evolution of the specificity of the peptide receptor
Ste2 in the phylum Ascomycota. We identified Ste2-like receptors
from different species and expressed a subset of them in S.
cerevisiae. Thirteen of the heterologously expressed receptors
successfully responded to self-pheromone and some of them are
surprisingly promiscuous and can respond to high concentrations of
pheromones from different species. We see no evidence of positive
selection in the pheromone receptors contrasting with previous
studies of genes involved in speciation, and this can explain the
cross-talk between different receptors and pheromones.
We tested whether this lack of specificity could be enough to allow
detection, recognition and mating. We crossed two sympatric species,
S. cerevisiae and S. castellii and observed that they can fuse at high
frequency but no viable diploids are formed in the process.
We conclude that in the Saccharomyces, speciation is most likely not
happening at the receptor/pheromone recognition level and that, in
these species, strong post-zygotic isolation is not sufficient to prevent
mating.
14

Regulation of tissue growth in Drosophila
Paulo S. Ribeiro
Postdoctoral Fellow
Tapon Lab - Apoptosis and Proliferation Control Laboratory
Cancer Research UK, London Research Institute
44 Lincoln’s Inn Fields
London WC2A 3LY
United Kingdom
Metazoan organ development requires strict control of cell
proliferation, growth and death. Studies in Drosophila have identified
a novel signalling pathway, the Hippo (Hpo) pathway, which is
required for tissue size control as it inhibits proliferation and
promotes apoptosis. Since many of the pathways that limit tissue size
during development also function in adults, they are often the targets
of cancer-causing alterations. Hpo pathway members are highly
conserved and, depending on their function, can act as tumour
suppressors or oncogenes in mammals.
The Hpo pathway consists of a kinase cascade comprising the kinases
Hpo and Warts (Wts), which culminates in the inhibition of Yorkie
(Yki), a growth-promoting transcriptional co-activator involved in
tumourigenesis. Hpo signalling is also regulated by upstream
members, such as the atypical cadherin Fat, the cytoskeletal band
4.1 proteins Expanded (Ex) and Merlin (Mer), and the newly identified
Kibra. Moreover, the kinase adaptor proteins Mats, Salvador (Sav)
and dRASSF can also control the activity of the pathway. Although
some of the major pathway components are known, the molecular
mechanisms regulating Hpo signalling activity are still unclear. In
particular, little is known about the negative regulation of the kinase
cascade that lies at the core of the Hpo pathway.
Using Drosophila as a model system, we have recently characterised
the in vivo and molecular functions of a PP2A phosphatase complex
that negatively regulates Hpo signalling, which was identified using
combined high-throughput genomic and proteomic approaches.
We will discuss these new findings and the future of Hpo pathway
research.
15

Laboratory Orders: how to enjoy 2 extra weeks of vacation
Tiago P. Carvalho
PetriDish Software Lda.
After completing his PhD in 2009, Tiago decided to return to Portugal
to start "something" there. That "something" is now PetriDish
Software Lda., a software company located at the Science and
Technology Park of the University of Porto (UPTEC).
The name of the company is homage to Julius Richard Petri, who
invented the Petri dishes, while he was working as a research
assistant in the laboratory of the not-less-famous Robert Koch. Petri
dishes are a simple but brilliant invention, and their quick adoption
made some laboratory practices so straightforward that today one
rarely thinks about the difficulties that existed before Petri dishes
were invented.
Similarly, PetriDish Software is devoted to the development and
implementation of simple but smart software solutions. We humbly
aim at making a Scientist\'s life so much easier, to the point that one
day no one will understand how it would be possible to work without
some of our software solutions.
Ordering laboratory products or reagents is perceived by the
Scientific community as an annoying and time-consuming process.
Our first product is the LabOrders online platform, which makes that
process easier, faster, cheaper and more efficient (at
http://www.laborders.com).
In this talk I will present LabOrders live, while sharing the UPs and
DOWNs of starting a company in Portugal
16

The obscurity of plasmalogens in the spotlight
Pedro Brites
Nerve Regeneration Group, Instituto de Biologia Molecular e Celular,
Porto, Portugal
The importance of peroxisomes for normal function and physiology is
highlighted by the severe disease sate, with severe pathology and
poor outcome of patients with a disorder caused by impaired
peroxisomal function. Peroxisomes perform a myriad of functions that
include beta-oxidation of very-long-chain fatty acids, alpha-oxidation
of phytanic acid, degradation of hydrogen peroxide, and the
biosynthesis of plasmalogens (PLS).
The peroxisomal disorder, rhizomelic chondrodysplasia punctata
(RCDP) is caused by the impairment in the biosynthesis of PLS and is
characterized by defects in bone, eye and nervous tissue. PLS are
ether-phospholipids characterized by a vinyl-ether bond at the sn-1
position of the glycerol backbone. Despite an unclear physiologic
function, PLS function as (i) determinants of membrane structure and
fluidity, (ii) pools of polyunsaturated fatty acids and (iii) scavengers
of oxidative damage. Using mouse knockout models deficient in PLS
we aim at understanding human disorders, characterize the defects
caused by a PLS deficiency and determine the mechanism and
players involved in pathology caused by PLS deficiency. Since
nervous tissue is extremely rich in PLS we set out to characterize and
determine the role of PLS in neurons and in myelin. Our analysis
revealed that in neurons a deficiency in PLS causes abnormalities in
the structure and patterning of neuromuscular junctions.
A deficiency in PLS also leads to abnormalities in myelin and in
myelination characterized by decompaction of myelin sheaths and
impaired myelination during development, and after a lesion. Our
results indicate that PLS are important structural components of
membranes and that their deficiency affects multiple processes. Due
to their role in membranes, PLS may also modulate the disease state
of non-peroxisomal disorders.
17

At the Heart of INEB's Stem Cell Biology Team
D.S. Nascimento, A. Freire, M. Valente and P.Pinto-do-Ó
NEWTherapies Group, INEB - Instituto de Engenharia Biomédica,
Universidade do Porto, Porto, Portugal
Our team focus is on elucidating how stem/progenitor cells (SC)
engage in regeneration and repair in adult-tissues. Special emphasis
is placed on the mechanisms involved in the regulation of the socalled
SC niches. Currently, the working paradigm is that of the heart
as an organ endowed with endogenous cardiac stem/progenitor cell
(CSC/CPC) population(s) and thus with self-regenerating potential.
Myocardium-resident putative CPCs display hallmarks of stemness
and their antigenic profile, i.e. Sca+/c-Kit+/MDR1+, by matching that
of other adult SC, raises the question as to whether CPCs originate in
the heart or are continuously replenished from other organs. We
address these issues in an integrative manner. An experimental
mouse-model of myocardial infarction (MI) was established and a
thorough cellular/molecular analysis of specific CPC-subsets is
underway. Aiming at the identification of molecules involved in CPCs
stress-response signaling pathway(s), a series of transcripts have
been serially evaluated in the Sca-1+ CPCs isolated from nonmanipulated,
sham-operated and mice subjected to MI. At the tissuelevel,
and to portray the CPCs, their differentiating progeny and
supporting cells at the natural niche, protocols/tools for optimal in
situ analysis have been a subject of continuous investment. Thus,
from the digging and drawing of the cardiac stem/progenitor cells
response(s) to heart injury a draft of (i) the kinetics of the cardiacniche’s
composition and of (ii) the molecular blueprint of the CPC
response to MI starts to emerge.
18

From evolution to the bed-side
Jose B. Pereira-Leal
Instituto Gulbenkian de Ciência – Portugal
Whether translational research should go from mechanistic cellular
and molecular studies to its application, i.e. bench to the bed-side, or
from the clinical problem and human samples to the basic laboratory,
i.e. bed-side to the bench, is an issue that polarizes the biomedical
community. I will discuss an example of a less explored translational
avenue - from evolutionary studies to the bed-side. In the
Computational Genomics Laboratory (evocell.org) of the Institute
Gulbenkian de Ciência we are broadly interested in the evolutionary
cell biology of cellular compartmentalization or modularity. We aim to
understand the evolutionary mechanisms by which cellular processes
are physically separated, focusing on intracellular organelles. While
studying the origin of bacterial-derived organelles (e.g. mitochondria,
chloroplasts, etc) we investigated the drivers of gene loss in
intracellular parasites. We discovered that in addition to well-known
drivers, e.g. no need for biosynthetic pathways, there is extensive
loss of genetic redundancy.
From an evolutionary perspective, our results support the notion that
environmental predictability is a major driver of genome evolution.
From a translational perspective, this discovery underpinned a new
approach to drug repositioning, i.e. finding new uses for existing
drugs, that already uncovered a new type of antibiotic. I will also
discuss how other “evolutionary” projects in the lab are leading us to
a translational context.
19

Fitness Landscape of a Metabolic Pathway
Lilia Perfeito (1,2) Stephane Ghozzi (2) Johannes Berg (2) Karin
Schnetz (1) and Michael Laessig (2)
(1) Institut für Genetik, Universitat zu Köln, Zälpicherstrasse 47a,
50674 Cologne, Germany
(2) Institut für Theoretische Physik, Universitat zu Köln,
Zälpicherstrasse 77, 50937 Cologne, Germany
In order to understand how new functions evolve, it is important to
know which phenotypes and under selection and how mutations
affect them. In this work, I show how this approach can be used to
address the problem of the evolution of gene regulation. Genes are
regulated because their expression involves a fitness cost to the
organism. The production of proteins by transcription and translation
is a well-known cost factor, but the enzymatic activity of the proteins
produced can also reduce fitness. We mapped the fitness costs of a
key metabolic network, the lactose utilization pathway in Escherichia
coli. We measured the growth of several regulatory lac operon
mutants in different environments inducing expression of the lac
genes. We find a strikingly nonlinear fitness landscape, which
depends on the production rate and on the activity rate of the lac
proteins.
A simple fitness model of the lac pathway, based on elementary
biophysical processes, predicts the growth rate of all observed
strains. The nonlinearity of fitness is explained by a feedback loop:
production and activity of the lac proteins reduce growth, but growth
itself reduces the density of these molecules through dilution. This
nonlinearity has important consequences for pathway function and
evolution. It generates an activation threshold of the lac operon,
above which bacterial growth abruptly drops to low values.
Furthermore, the nonlinearity determines how the fitness of operon
mutants depends on the inducer environment. Fitness nonlinearities,
growth thresholds, and gene-environment interactions are likely
generic features of metabolic pathways and have implications for the
evolution of regulation.
20

NFκB signaling cooperates with cMyc to induce B cell transformation
and multiple myeloma in the mouse
Calado DP (1) Zhang B (1) Sasaki Y (2) Wunderlich T (3)
Schmidt Supprian M (4) and Rajewsky K (1)
(1) PCMM/Immune Disease Institute, Harvard Medical School, Boston,
USA
(2) RIKEN Center for Developmental Biology, Kobe, Japan
(3) The Institute of Genetics of the University of Cologne, Cologne,
Germany
(4) Max Planck Institute of Biochemistry, Munich, Germany
Constitutive NFκB signaling and deregulation of cMyc expression are
recurrent events in the ABC subgroup of Diffuse Large B cell
Lymphoma (ABC/DLBCL), and in Multiple Myeloma (MM). However, it
remains to be determined whether constitutive NFκB signaling and
deregulated cMyc expression, cooperate to induce B cell
transformation and consequently are at the origin of ABC/DLBCL and
or MM. To test this hypothesis we used conditional targeted
mutagenesis in the mouse, to express a constitutively active IKK2
molecule and/or cMyc in CD19pos B cells through tamoxifen induced
Cre mediated elimination of a STOP cassette. The genetic system of
conditional targeted mutagenesis used in this work is unique in two
ways. First, transgene activation occurs only in a small fraction of
CD19pos B cells (3 to 5%) through transient Cre expression,
mimicking the sporadic nature of oncogenic events. Second, due to
incomplete Cre mediated recombination in individual cells, this
CD19+ B cell population is constituted by IKK2capos/MYCpos,
IKK2capos/MYCneg and IKK2caneg/MYCpos expressing cells,
providing all possible “oncogene” combinations in the same mouse at
the same time.
We observed that 100% of the mice succumbed to a plasma cell
neoplasia derived from the IKK2capos/MYCpos population, with a
median survival of 192 days. Thus, using a conditional targeted
mutagenesis system that recapitulates the sporadic nature of
oncogenic events, we demonstrate that constitutive NFκB signaling
cooperates with deregulated cMyc expression to induce B cell
transformation and MM in the mouse. These results suggest that this
oncogene combination represents a primary event in MM rather than
ABC/DLBCL.
21

Tight Junction components: novel players in Epithelial
Morphogenesis?
Ana Cristina Borges (1,2) Ana Cristina Silva (1,2) Jorge Carneiro
(1) Mathias Koeppen (1) and António Jacinto (1,2)
(1) Instituto Gulbenkian de Ciencia
(2) Instituto de Medicina Molecular
We are interested in the understanding of the molecular mechanisms
involved in the sealing of an epithelial discontinuity that occur both in
normal development and eventually during tissue repair. We are
using the zebrafish embryo as a model system, since it allows the
study of the morphogenetic movements of epiboly and wound
healing. In both cases, closure of the gap is achieved as the opposing
tissue margins move toward each other to undergo fusion. Tissue
movement is driven by coordinated changes in cellular morphology,
which require mechanical force. How is such force generated in a
moving sheet of cells? A common feature observed both in normal
morphogenesis and wounds is the formation of a supracellular
actin/myosin cable which contracts and closes the gap in a purse
string-like fashion.
Currently, we are investigating the role of Tight Junctions (TJ) in
these processes. Our preliminary results revealed that the TJ protein
tjp-3/ZO-3 is required for the formation of the actin/myosin cable.
The impairment in cable formation leads to a delay of the epiboly
process and to a failure in wound closure. These observations
implicate TJ components as novel players in epithelial morphogenesis.
22

How to preserve vision? Path to clinical application using human
neural stem cells
Alexandra Capela
StemCells Inc.
Age-related macular degeneration (AMD) and retinitis pigmentosa
(RP) are the two most prominent human retinal degenerative
disorders and combined account for the majority of vision loss in
people over 40 years old.
StemCells Inc. has developed a purified population of human central
nervous system stem cells (hCNS-SCns) that are currently being
tested in 3 clinical studies, Neuronal Ceroid Lipofuscinosis, a
lysosomal storage disorder, Pelizaeus-Merzbacher Disease, a
dysmyelination disorder and Spinal Cord injury. We now demonstrate
that hCNS-SCns transplanted into the subretinal space of the Royal
College of Surgeons (RCS) rat model, a well-characterized animal
model of progressive photoreceptor degeneration, rescue
photoreceptors from degeneration and preserve visual acuity and
visual field sensitivity. These findings support clinical evaluation of
hCNS-SCns as a neuroprotective strategy for select retinal
degenerative disorders.
23

p53-targeted tumor therapy: potential and limitations
Melissa R. Junttila, Daniel Garcia, Anthony N. Karnezis, Francesc
Madriles, Gerard I. Evan and Carla P. Martins(*)
University of California San Francisco, Department of Pathology and
Helen Diller Family Comprehensive Cancer Center, San Francisco,
California 94143-0502, USA
(*) Present address: Cancer Research UK Cambridge Research
Institute, Li Ka Shing Centre, Robinson Way, Cambridge CB2 ORE, UK
The high prevalence of specific mutations in tumors made targeted
therapies a new priority in drug development. Inactivation of the p53
pathway is a common feature in human tumors fostering the
attractive notion that p53-function restoration would constitute an
effective and tumor-specific therapeutic strategy. To establish the
therapeutic potential of this approach we modeled in vivo the impact
of p53 restoration in tumors. We recently showed that p53
restoration triggers dramatic tumor regression in a murine lymphoma
model, highlighting the therapeutic potential of this strategy. To
determine its impact in epithelial tumors we carried out a
comprehensive analysis of the timing and mechanisms of p53
activation during the evolution of lung tumors, the leading cause of
cancer-related death worldwide. Using a murine model of non-small
cell lung carcinoma we show that unexpectedly, p53 restoration failed
to induce lung tumor clearance. Nevertheless we show that p53
efficiently targets the most malignant tumor cells, but allows for the
less aggressive cell populations to survive. This tumor stage specific
activation of p53 results from a selective increase in oncogenic
signaling in malignant cells. The failure of low-level oncogenic stress
to engage p53 reveals inherent limits in the capacity of p53 to
restrain early tumor evolution and in the efficacy of therapeutic p53
restoration to eradicate cancers. Strategies aimed at improving the
therapeutic impact of targeted therapies for lung cancer treatment
will also be discussed.
24

The remarkable functional divergence of the Eukaryotic organellar
Release Factors
Isabel Duarte, Ramiro Morgado and Martijn Huynen
ID and MH are affiliated with CMBI - NCMLS, Radboud University,
Nijmegen, The Netherlands; RM is affiliated with Computational and
Systems Biology - JIC, Norwich, UK
Motivation: Organellar gene expression is far from understood, with
the genetic code of the human mitochondrion being elucidated only in
2010.
Two codon-specific RFs are sufficient for de facto organellar
translation termination: RF1 and RF2, but at least 3 other different
subfamilies have been described, whose exact function remains
elusive. Our detailed analysis sought to integrate different sources of
information: localization prediction and experimental data available,
phylogenetic distribution, organellar genetic code and sequence
structural features, to better describe this superfamily and predict
unannotated proteins function.
Materials and Methods: We used comparative genomics (Sequence
Homology Searches, Multiple Sequence Alignment and Phylogeny)
and Protein Structure Prediction to study the organellar RF
superfamily in a carefully selected group of representative Eukaryotic
organisms.
Results and Discussion: This study briefly reports a new plant specific
release factor subfamily, which has lost the 3 functional motifs
experimentally described as essential for bona-fide release factor
activity. Also, it predicts a plant RF2 subfamily, to be chloroplastic
and not mitochondrial as previously annotated. And finally, we
propose a possible re-invention of the standard genetic code in the
green algae lineage.
Conclusion: Here we analyze the evolution of organellar ribosomal
Release Factors, uncovering a remarkable functional differentiation
and showing its co-evolution with the organellar genetic code. Also,
our studies confirm the previously published evolutionary origin for
the mitochondrial and plastidial RFs, in accordance with the currently
undisputed endosymbiotic origin of these organella. Finally, the motif
shuffling and loss seen on each RF subfamily, hints at a significant
functional divergence, hence proving that much is still to be
uncovered about this process!
25

ABSTRACTS FOR POSTER PRESENTATIONS
Monday 27 December
SESSION TWO
26

Centrosome & Cilia Biogenesis & Evolution
Mónica Bettencourt-Dias
Instituto Gulbenkian de Ciência
Our research focuses on cell cycle progression and the cytoskeleton in
normal development and disease. We are particularly interested in
the role played by microtubule organizing structures, such as the
centrosome, cilia and flagella. The centrosome is the major
microtubule organizer in animal cells, and is very often abnormal in
cancer. Cilia and flagella are cellular projections which are
indispensable in a variety of cellular and developmental processes
including cell motility, propagation of morphogenic signals and
sensory reception. Despite their importance, we know very little
about centrosome and cilia biogenesis or how they may go awry in
human disease. Our laboratory uses an integrated approach to study
those questions: we combine studies in model organisms with studies
in human cells and bioinformatics to have an integrated view of this
process. The fruit fly is an excellent organism to address those
questions, since it combines possibilities of screening
multiple genes with the ability to perform in-depth regulation studies
in the whole organism. As the regulatory mechanisms of the cell cycle
and cytoskeleton have been highly conserved throughout evolution,
we can extrapolate our findings to humans to test their relevance for
human disease. An understanding of the pathways involved in cell
cycle and cytoskeleton can generate diagnostic and prognostic
markers and hopefully provide novel therapeutic targets in human
disease.
28

Shared mechanisms during neural development and tubulogenesis
Sofia J. Araújo
IBMB-CSIC, IRB Barcelona, Spain
Tissue and organ morphogenesis in multicellular organisms relies on
the precise coordination of multiple molecular pathways that control
distinct cellular events. Cellular migration and pathfinding are two
processes essential for morphogenesis. Their precise temporal and
spatial coordination is crucial for cells to be able to establish
connections over long distances. Correct control allows development
of multicelular organisms. Defects in control are associated with
cancer development.
In Drosophila melanogaster neural and tracheal morphogenesis both
involve cellular migration and pathfinding. To date, studies of
Drosophila’s nervous and tracheal development have been often
performed separately, despite the many common features between
the two systems. This overlap is suggestive of shared mechanisms
and cellular cooperation are likely to exist. We have been
investigating the cooperative influence of tracheal cell migration and
axonal guidance during development. We have analysed a genetic
screen for which mutants were isolated and characterised by their
nervous system phenotypes and investigated their tracheal
phenotypes. This approach is allowing us to identify novel factors
involved in the development of both systems.
29

Characterization and applications of a novel hemangioblast-specific
transcriptional enhancer
Ana Teresa Tavares (1,2) Vera Teixeira (2) and António Duarte
(1,2)
(1) CIISA, FMV, Lisboa, Portugal
(2) IGC, Oeiras, Portugal
During early vertebrate embryogenesis, there is a close
developmental relation between hematopoiesis and vasculogenesis.
Primitive hematopoietic and endothelial lineages both derive from
aggregates of mesodermal cells that form the blood islands in the
extraembryonic yolk sac. This observation has led to the hypothesis
that both cell types originate from a common precursor known as the
hemangioblast. Chick Cerberus gene (cCer) codes for a secreted
factor expressed in the anterior mesendoderm that gives rise to blood
islands, among other tissues.
During the study of cCer transcriptional regulation in chick embryos,
we isolated a short cis-regulatory region that is able to drive reporter
gene expression specifically in blood island precursor cells or
hemangioblasts. We have been using this hemangioblast-specific
reporter construct (Hb-eGFP) to characterize the gene expression
profile of the chick hemangioblast and to track the origin and fate of
hemangioblasts in living embryos. Here, we will present data on the
cell type-specificity of the hemangioblast enhancer as well as on the
dynamics of blood island morphogenesis and differentiation observed
in Hb-eGFP-electroporated chick embryos.
30

A genetic screen to identify genes mediating ER stress induced retinal
degeneration in Drosophila
Vanya Rasheva, Nadine Schweizer and Pedro Domingos
Instituto de Tecnologia Química e Biológica (ITQB), Universidade
Nova de Lisboa, Av. Republica (EAN), 2780-157, Oeiras, Portugal
(domingp@itqb.unl.pt)
The endoplasmic reticulum (ER) is the cell organelle where secretory
and membrane proteins are synthesized and folded. Proteins that fail
to fold properly (misfolded proteins) accumulate in the ER, causing
ER stress. The Unfolded Protein Response (UPR) consists of several
signalling pathways that have evolved to detect the accumulation of
misfolded proteins in the ER and activate a cellular response that
attempts to maintain homeostasis in the ER. If the protective
mechanisms activated by the UPR are not sufficient to restore normal
ER function cells die by apoptosis. How is the UPR regulated to
change from a protective response to a pro-apoptotic one is still
under question.
We are performing a mosaic genetic screen aimed at identifying
suppressors of the retinal degeneration phenotype caused by Xbp1
overexpression in the eye. The identification of genes regulated by
Xbp1 will allow the characterization of the protective and apoptotic
signalling pathways induced by ER stress.
31

WDR62 in microcephaly and assymetric brain development
Ofelia P. de Carvalho, Adeline K. Nicolas, Maryam Khurshid and C.
Geoffrey Woods
Cambridge Institute for Medical Research, University of Cambridge
Autosomal recessive primary microcephaly (MCPH) is a disorder of
neurodevelopment resulting in a small brain. We identified
homozygous and frame-shifting deletions of WDR62 in seven
consanguineous MCPH families. As well as being involved in MCPH,
WDR62 mutations have been identified in families with a wide
spectrum of brain development abnormalities including but not
exclusive of microcephaly.
In human cell lines WDR62 localizes to the spindle pole during
mitosis, and in both mouse and human embryonic brain expression is
found in neural precursors during mitosis. We attempt to elucidate
the developmental mechanism(s) perturbed by mutations in this
gene.
32

Mutualistic Parasites - when hosts make use of their parasites and
pathogens
Francisco Dionisio
Centro de Biologia Ambiental da Faculdade de Ciencias da
universidade de Lisboa & Instituto Gulbenkian de Ciencia
Parasites or pathogens and their hosts are expected to have
conflicting interests, simply because parasite reproduction and
transmission is done at the expense of the hosts fitness. However,
this may not always be the case. Infected hosts can use their
parasites/pathogens to harm others, and increase their own relative
fitness.
33

The impact of antiretroviral treatment on the burden of invasive
pneumococcal disease in South African children: A time series
analysis
Marta C Nunes, Shabir A and Madhi
Respiratory and Meningeal Pathogens Research Unit, Department of
Science and Technology/National Research Foundation: Vaccine
Preventable Diseases, University of the Witwatersrand, South Africa
Objective: HIV infection is a major risk factor for invasive
pneumococcal disease (IPD). A national antiretroviral program was
initiated in South Africa in 2004. This study evaluates the impact of
the highly active antiretroviral (HAART) treatment program on the
burden of IPD among African children.
Design: Retrospective analysis of laboratory-confirmed IPD among
children under 18 years of age, from 2003 to 2008.
Methods: The periods 2003-2004, 2005-2006 and 2007-2008 were
defined as the early-, intermediate- and established-HAART eras,
respectively.
Pneumococcal conjugate vaccine was not introduced into public
immunization during this period.
Results: 1,171 episodes of IPD were identified over the study-period.
Among HIV-infected children under 18 years, the burden of IPD
decreased by 50.8% (95% CI: 41.5; 58.7) and the incidence of IPDrelated
mortality declined by 65.2% (95% CI: 47.2; 77.0) from the
early- compared to the established-HAART era. This decline in HIVinfected
children was evident for pneumococcal bacteremia and
pneumococcal meningitis. In addition, similar reductions were
observed for serotypes included in a 7-valent pneumococcal
conjugate vaccine and non-vaccine serotypes. The burden of IPD
remained unchanged in HIV-uninfected children under 18 years of
age over these periods. The risk of IPD, however, remained 42-fold
greater in HIV-infected compared to HIV-uninfected children in the
established-HAART era.
Conclusions: Although the HAART program has been associated with
significant declines in IPD morbidity and mortality, HIV-infected
African children with access to HAART remain a high-risk group for
IPD. These children should therefore be prioritized in the prevention
of IPD.
34

Role of Dll4-Notch signaling pathway in the angiogenesis of
atherosclerotic lesions and its potential therapeutic aplication
Vera Teixeira (1,2) Joana Gigante (1,2) Dusan Djokovic (1,2) and
Ana Tavares (1,2)
(1) CIISA, Faculdade de Medicina Veterinaria, Lisboa
(2) Instituto Gulbenkian de Ciencia, Oeiras, Portugal
Atherosclerosis is a degenerative disease characterized by vascular
inflammation, endothelial dysfunction and progressive formation of
intima plaques composed of lipids, calcium, and cellular debris. As
plaque develops, new vessels arising from the adventitial vasa
vasorum grow into the media and intima lesions. Evidences implicate
these neovessels in the regulation of plaque growth and stability.
Atheroma neovascularization inhibition has been proposed as an
approach to restrict plaque growth and promote stabilization. Indeed,
a promising antiangiogenic strategy for atherosclerosis would
decrease neovessel proliferation and induce functional maturation of
the plaque microvessels. However, preliminary observations showing
Dll4 expression increased in atheroma suggest also therapeutical
benefit in Dll4/Notch-inhibition. Therefore, we will investigate Dll4
expression pattern during plaque development, vasa vasorum levels
in ApoE-/- mice and human patients, function of Dll4 in plaque
angiogenesis and the therapeutic potential of Dll4/Notch signaling
modulators in the prevention and/or treatment of atherosclerosis.
Taken our current knowledge on the function of Dll4 in physiological
and tumor angiogenesis we expect modulation of Dll4 levels to be an
effective treatment of atherosclerotic lesions. Our findings will
contribute to unravel the molecular mechanisms of atheroma
angiogenesis.
35

Activity Dependent Cleavage of Neuroligin-1
Rui Peixoto, Portia McCoy, Ben Philpot and Michael Ehlers
Duke University, University of North Carolina
Adhesive contact between pre- and postsynaptic neurons initiates
synapse formation during brain development and provides a natural
means of trans-synaptic signaling. Numerous adhesion molecules and
their role during synapse development have been described in detail.
However, once established, the mechanisms of adhesive disassembly
and its function in regulating synaptic transmission have been
uncertain. Here, we report that synaptic activity induces acute
proteolytic cleavage of neuroligin-1 (NLG1), a postsynaptic adhesion
molecule at glutamatergic synapses. NLG1 cleavage is triggered by
NMDA receptor activation, requires Ca2+/calmodulin-dependent
protein kinase, and is mediated by proteolytic activity of matrix
metalloprotease 9 (MMP9). Cleavage of NLG1 occurs in vivo, is
regulated over brain development, and causes rapid destabilization of
its presynaptic partner neurexin-1β (NRX1β). In turn, NLG1
cleavage rapidly depresses synaptic transmission by abruptly
reducing presynaptic release probability. Together, our results define
a novel mechanism for synapse remodeling and trans-synaptic
signaling during brain development and plasticity.
36

Ensembl Regulation: Functional Genomics @ Ensembl
N. Johnson, D. Sobral, D. Keefe, B. Pritchard, S. Wilder, E. Birney, P.
Flicek and I. Dunham
European Bioinformatics Institute (EMBL-EBI), Wellcome Trust
Genome Campus, Hinxton, Cambridge, UK
The Ensembl Functional Genomics (eFG) database and Application
Program Interface (API) provides a platform for storage, analysis and
visualisation of functional genomics data, within the centralised family
of EnsEMBL databases. Widespread uptake of Nexte Generation
Sequencing (NGS) based methods in projects including HEROIC,
ENCODE and the Epigenomics Roadmap, as well as smaller
hypothesis-driven projects, has initiated a flood of data on
transcription factor (TF) binding and chromatin state. The eFG
databases incorporate 285 data sets from these projects, primarily
from ChIP-seq and DNase-seq assays across 9 human and 4 murine
cell lines. This includes the genomic binding sites for 56 TFs as well
as the locations for 40 histones modifications. An additional 23 data
sets identifying sites of open chromatin or DNase 1 hypersensitivity
are also now available. Data is incorporated via a standardised read
mapping and peak calling pipeline to generate both normalised
signal data and predicted enriched regions (“peaks” or “hits”) with
peak summits. Both signal data and enriched regions are made
available in the Ensembl browser for visual comparison and
interrogation.
Higher-level annotations are also available in eFG, in the form of
‘Regulatory Features’. The ‘Regulatory Build’ process uses all TF and
open chromatin data across all cell types to establish locations that
are potentially active in regulation (regulatory cores) in a multi-cell
build step. Each cell-specific core region is then extended, given that
appropriate supporting data (e.g. histone markers) is present in that
cell type. Regulatory Features are then classified by considering how
different patterns of attributes are distributed in relation to various
genomic features (e.g. promoter-associated, gene-associated). This
process gives a set of annotated Regulatory Features for each cell
type, as well as a set of core Regulatory Features presented in a
multi-cell track.
Ensembl is committed to immediate release of data and analysis into
the public domain. As with all Ensembl software, eFG is available
under an open source license from www.ensembl.org.
37

Notch signalling regulation by cis-inhibitory effects
Ulla-Maj Fiuza, Isabelle Becam,Thomas Klein, Marco Milán and
Alfonso Martinez Arias
Department of Genetics (University of Cambridge)
Institute for Research and Biomedicine (Barcelona)
Institute for Genetics (University of Cologne)
Notch signalling is used iteratively during development and also in
the adult organism. There are several known diseases related with
miss-regulation of the signalling activity, many causing severe
defects and some fatal. Understanding the mechanisms involved in
the regulation of this signalling pathway is of great importance for
addressing how to tackle Notch related pathologies. In this work we
present a study on the mechanisms of Notch signalling regulation
mediated by interactions between the Notch receptor and its ligands
Delta and Serrate. We start by further characterizing a well
established observation that Notch ligands besides having an
activating role can have an inhibitory activity in a concentration
dependent manner. This function of ligands has been many times
ascribed in the literature as a cell autonomous effect but no formal
evidence for this had been previously produced. We further present in
vivo evidence suggesting that the Notch region involved in this
inhibitory interaction is the Notch EGF-repeats 10-12 domain and cell
culture data indicating that the ligand-inhibitory effect results from
high levels of ligand inhibiting the ectodomain shedding of Notch.
Another type of inhibitory interactions is also explored that Notch can
have a cis-inhibitory activity on Serrate signalling ability. The
mechanism behind this regulatory interaction is identified as an
inhibitory regulation by Notch of the levels of Serrate. These crossregulatory
interactions have paramount importance in establishing
the state of signalling activity in different biological contexts for
example: ligand cis-inhibition has been seen to modulate stem cell
maintenance in epithelial tissues and there are now evidences
towards evolution leading to protein specialization with some Delta
family molecules only having an inhibitory activity. It will be
interesting to see how this regulatory mechanisms act in multiple
biological contexts.
38

The SR45 plant-specific splicing factor is a negative regulator of sugar
signaling during early growth in Arabidopsis
Raquel Carvalho, Sofia Carvalho and Paula Duque
Instituto Gulbenkian de Ciência, Oeiras, Portugal
Being sessile, plants have developed unique adaptive strategies to
cope with abiotic stress. These range from morphological
modifications to physiological responses at the cellular level, but the
basis of the capacity for adaptation lies ultimately at the level of the
genome. Many plant genes undergo alternative splicing, which
increases transcriptome and proteome complexity likely to be crucial
in plant tolerance to adverse environmental conditions.
Serine/arginine-rich (SR) proteins are known to modulate alternative
splicing and other aspects of RNA metabolism, but their potential role
in plant stress responses remains unexplored. Using reverse genetics,
we found that SR45, a plant-specific Arabidopsis SR protein highly
conserved throughout the plant kingdom with no orthologs in
animals, is a novel player in glucose signaling during early seedling
development.
The sr45-1 mutation confers hypersensitivity to glucose, with mutant
seedlings grown in the presence of the sugar displaying impaired
cotyledon development and hypocotyl elongation, as well as altered
glucose-responsive gene expression. The mode of action of SR45
involves downregulation of the abscisic acid (ABA) stress signaling
pathway, via repression of both glucose-induced ABA endogenous
accumulation and ABA biosynthesis and signaling gene expression.
Epistasis analyses suggest a mechanism independent of the
hexokinase 1 (HXK1) sugar sensor, and preliminary results indicate
that SR45 modulates the levels of KIN10, a protein kinase involved in
sensing/signaling of stress-associated energy deprivation.
39

When knowledge is the product – what is customer value?
A study of market orientation in a research not-for-profit
organisation.
Greta Martins
MBA dissertation thesis, University of Leicester, UK
The rationale of market orientation (MO) is based on Porter (1985)’s
five-competitive force model for industries in the creation of value by
raising entrance barriers through engendering and sustaining
competitive advantage. The marketing philosophy holds that a way to
do this is by satisfying customer needs and wants more effectively
than competitors. MO is conceptualised on profitability maximisation
through delivering superior customer value to competitors by
harnessing resources on an organisation-wide basis, hence literature
advocates the existence of an MO culture in an organisation.
Rapidly changing environments have forced not-for-profits (NFP)s to
adopt businesses practices such as MO for sustainability and growth.
Indeed studies show that NFPs favour stronger links with
organisational performance than businesses in developing an MO
strategy. However, studies reveal that due to the higher social
context of the NFP environment, as opposed to mandatory initiatives,
it is more effective to adopt market activities into the culture so as to
develop a market orientation culture. This study takes a counter
view that when a NFP is knowledge-based, MO is developed through
organisational culture. Therefore the nature of the NFP activity
influences the development of MO.
A small NFP scientific research institution (the Instituto Gulbenkian de
Ciência) was selected as empirical context. Semi-structured
interviews were conducted to gain insight into the development of MO
through the relationship of the scientists with a key stakeholder
group – the PhD students. In considering the students as customers
what were the perceptions of customer value?
40

ABSTRACTS FOR ORAL PRESENTATIONS
Monday 27 December
SESSION THREE
42

Host response mechanisms to malaria liver infection: The role of
ubiquitin-proteasome pathway
Zuzarte-Luís (1) N. Nagaraj (2) C. Carret (1) M. Mann (2) and M.
Mota (1)
(1) Instituto de Medicina Molecular, Portugal
(2) Max Planck Institute for Biochemistry, Martinsried, Germany
Malaria is caused by Plasmodium, an apicomplexan parasite
transmitted through the bite of an infected female Anopheles
mosquito. The infection starts when the mosquito injects Plasmodium
sporozoites into the skin of the vertebrate host. After traversing the
dermis the parasites enter the circulatory system and travel to the
liver where they undergo major transformations and commence a
period of remarkable schizogynous division into an exoerythrocytic
form (EEF). This asymptomatic development step will eventually lead
to the release of thousands of mature merozoites into the
bloodstream, initiating the symptomatic stage of the disease.
In humans, the hepatic stage of the infection lasts 6-10 days during
which the number of parasites expands enormously, up to 40000
fold. Surprisingly, such an active process of parasitic development
progresses “invisible” to the host’s immune system. As a
consequence, immunity to the hepatic stage is not acquired naturally
and hence, populations in malaria endemic areas lack protective
immunity, even after repeated exposures to the parasite.
Understanding how the developing parasite overcomes the host
immune system guaranteeing its own survival will constitute a major
breakthrough, possibly leading to the development of an anti-malarial
vaccine.
44

Herpesviral Intrahost Spread
Miguel Gaspar and Philip Stevenson
University of Cambridge
Herpesviruses are important human pathogens able to establish a
life-long latent infection inside host cells hidden from the immune
system. Murid Herpesvirus 4 (MuHV4) is a gamma-herpesvirus that
infects mice and is commonly used as an in vivo model of the human
gamma-herpesviruses EBV and KSHV as the characteristics of their
infections are similar. MuHV4 enters the organism via the nose,
probably infecting an epithelial cell, and establishes latency inside B
lymphocytes in the lymph nodes and spleen. However, it is unknown
how the virus reaches the lymph nodes and spleen and how it infects
its main cellular target, B cells as these cells are resistant to infection
in vitro. We are currently testing the hypothesis that MuHV4 uses
dendritic cells (DCs) as Trojan horses to reach the lymph nodes and
to infect B cells by cell-to-cell transmission. We have already show
that DCs are susceptible to MuHV4 infection in vivo and in vitro and
that latent in vitro infection results in a dramatic rearrangement of
the actin cytoskeleton and other structural changes associated with
increased mobility.
We are currently using mice expressing Cre recombinase in DCs in
combination with viruses where loxP sites flank essential genes and
fluorescent markers in order to demonstrate the importance of DCs in
virus dissemination inside the host. The identification of a specific cell
type responsible for the transport of the virus from the site of lytic
replication to the site of latency can yield novel therapeutic targets to
prevent latency establishment.
45

Coupling RNA processing and chromatin modification in S.cerevisiae
Luis Soares and Steve Buratowski
BCMP, Harvard Medical School, Boston MA
Eukaryotic gene expression includes several steps that must be
coordinated to allow organisms to survive, develop and adapt to
environmental changes. The first step in gene expression involves the
transcription of genes with the production of mRNAs that are later
translated into proteins. Between the initial and final events of gene
expression the mRNA undergoes several processing steps that affect
its ability to be translated.
It has been shown that transcription and RNA processing are not
temporarily separated and must be connected in order to achieve
correct gene expression. Pre-mRNA processing events such as
splicing and poly-adenylation have been shown to be dependent on
the modulation of the activity of RNA polymerase II and on the
modifications of the CTD of its largest subunit.
Using S.cerevisiae as a model system we study the recruitment of
processing factors to transcribing genes and how this recruitment
correlates with chromatin modifications. Our results revealed that
modulation of particular histone modifications in yeast affect RNA
processing efficiency by modifying the cotrancriptional recruitment of
factors. We identified a novel pathway by which the COMPASS
complex (responsible for H3K4 methylation) affects the recruitment
of splicing and poly-adenylation factors during transcription. The
mechanism of this effect seems to be dependent both on the direct
recruitment of factors by this complex and on general effects that
H3K4 methylation has on the activity of RNA polymerase II.
46

Patterned cell-adhesion during morphogenesis
Ana Mateus and Alfonso Martinez Arias
Affiliations for Abstract - Department of Genetics, Cambridge
University, UK
Cell shape changes within epithelia require the regulation of adhesive
molecules that maintain tissue integrity. How remodelling of cell
contacts and maintenance of tissue integrity is achieved is a
fundamental question in morphogenesis. Cadherins are
transmembrane proteins that mediate cell-cell adhesion and, in
epithelia, localise at the adherens junctions (AJs). Analysis of
Drosophila E-Cadherin mutant embryos suggests that DE-Cadherin
regulation during Dorsal Closure (DC) relies mainly on posttranscriptional
regulation. In this study, we developed an assay to
assess the dynamics of DE-Cadherin. This assay revealed that native
DE-Cadherin has different antibody binding properties depending on
its location and the antibody used, which has never been observed in
fixed DE-Cadherin in Drosophila embryos. A monoclonal antibody
against the extracellular domain of DE-Cadherin labels amnioserosa
(AS) and dorsal-most epidermal (DME) cells, but not the lateral
epidermal cells, which contrasts with a polyclonal antibody that labels
all epidermal cells. These differences could result from different
conformation/organization of the Cadherins that confer different
adhesive strengths. Regional modulation of adhesion could represent
a general mechanism in morphogenesis, ensuring the maintenance of
adhesion in key regions of the tissue subjected to more tension.
47

Control of tissue growth and homeostasis by the integrated activities
of Hippo signaling and Drosophila Myc
Neto-Silva RM (1) (2) (*) and Johnston LA (2)
(1) Gulbenkian PhD Programme in Biomedicine, Instituto Gulbenkian
de Ciencia, Rua da Quinta Grande, 6, P-2780-156 Oeiras, Portugal
(2) Department of Genetics and Development, Columbia University
Medical Center, 701 West 168th Street, New York, NY 10032, USA
(*) Present address - Champalimaud Neuroscience Programme at
Instituto Gulbenkian de Ciencia, Rua da Quinta Grande, 6, P-2780-
156 Oeiras, Portugal
An understanding of how animal size is controlled requires knowledge
of how positive and negative growth regulatory signals are balanced
and integrated within cells. I will present data showing that the
activities of the conserved growth-promoting transcription factor Myc
and the tumor-suppressing Hippo pathway are codependent during
growth of Drosophila imaginal discs. We find that Yorkie (Yki), the
Drosophila homolog of the Hippo pathway transducer, Yap, regulates
the transcription of Myc, and that Myc functions as a critical cellular
growth effector of the pathway. We demonstrate that in turn, Myc
regulates the expression of Yki as a function of its own cellular level,
such that high levels of Myc repress Yki expression through both
transcriptional and posttranscriptional mechanisms. We propose that
the codependent regulatory relationship functionally coordinates the
cellular activities of Yki and Myc, and provides a mechanism of growth
control that regulates organ size and has potential implications for
cancer.
48

Slitrk5 a new neuronal protein implicated in obsessive-compulsivelike
behaviors in mice
Proenca CC, Shmelkov SV, Ninan I, Rafii S and Lee FS
Weill Cornell Medical College, New York
Instituto Gulbenkian de Ciencia, Oeiras
New York University Langone Medical Center, New York,
Obsessive-compulsive disorder (OCD) is a common psychiatric
disorder defined by the presence of obsessive thoughts and repetitive
compulsive actions, and often encompasses anxiety and depressive
symptoms as well. Recently, the cortico-striatal circuitry has been
implicated in the pathogenesis of OCD. However, the etiology and
pathophysiology of OCD is still not understood and the molecular
basis is not known. Several studies indicate that pathogenesis of OCD
has a genetic component. Here we demonstrate that loss of a
neuron-specific transmembrane protein, Slitrk5, leads to OCD
behaviors in mice, which manifests in excessive self-grooming and
anxiety, and is alleviated by the selective serotonin reuptake
inhibitor, fluoxetine. slitrk5-/- mice display selective abnormalities in
striatal anatomy and cell morphology, as well as alteration in striatal
glutamate receptor composition, which contribute to deficient corticostriatal
neurotransmission in adult mice. Thus, our studies identify
Slitrk5 as an essential molecule at the cortical-striatal synapses, and
provide a novel model to screen for pharmacological agents for the
treatment of OCD.
49

Forced NF-kappaB in T cells leads to Tumor Rejection
Cesar Evaristo (1) Thomas Gajewski (2) and Maria-Luisa Alegre (1)
(1) Department of Medicine
(2) Department of Pathology, University of Chicago, Chicago, IL
NF-kappaB activity has been reported to be reduced in T cells from
tumor-bearing hosts. Our previous results indicate that reduced NFkappaB
activation results in impaired survival of T cells, decreased
Th1 and Th17 differentiation and increased iTreg differentiation. Mice
with reduced T cell-NF-kappaB activity fail to reject cardiac and
pancreatic islet allografts in the absence of any pharmacological
treatment. We hypothesize that forced activation of NF-kappaB in T
cells should have the opposite effect and promote T cell survival,
facilitate Th1/Th17 differentiation and prevent iTreg differentiation,
which would be beneficial to reject tumors. We generated mice with
constitutively active IKKbeta in T cells (henceforth CA-IKKbeta mice).
Ectopic expression of CA-IKKbeta resulted in phosphorylation of NFkappaB.
Transgene expression was limited to CD4+, CD8+ and NKT
cells and T cells showed increased NF-kappaB activation and nuclear
translocation. T cell numbers were comparable to littermate controls,
but CA-IKKbeta mice had fewer Tregs and increased frequency of
activated T cells that produced IFNgamma upon re-stimulation. When
B16-SIY melanoma cells were injected subcutaneously, tumors grew
progressively in control littermates, whereas they were rejected by
mice expressing CA-IKKbeta in T cells. Furthermore, IKKbeta-CA
expressing CD4+ and CD8+ T cells were necessary and sufficient for
tumor control. Our results demonstrate NF-kappaB to be at the crossroads
of major T cell fate decisions that should uniquely synergize for
control of tumor growth.
50

Proteomics of Isolated Chromatin Segments
José Antão (1) James M. Mason(2) Jérôme Ájardin (3) and Robert
E. Kingston (1)
(1) Department of Molecular Biology, Massachusetts General Hospital,
Boston, USA
(2)NIEHS, Research Triangle Park, NC, USA
(3)Genetique Humaine, CNRS, Montpellier, France
The regulation of any genomic locus is done, in large measure, by the
complement of proteins that bind to it in a given cellular context.
Even though there are some powerful tools for looking at protein
localization in the genome through a candidate-based approach, an
unbiased method for identifying all proteins bound to a specific DNA
element at a given time has been lacking. We have developed a
method that uses a DNA/LNA-based oligonucleotide probe to target
and specifically purify genomic segments in their chromatin
environment, and determine their protein composition. Using this
method, we have identified proteins that bind to the Drosophila
Telomeric-Associated Sequence (TAS) repeats and, through a genetic
screen, we have confirmed the involvement of one of these proteins
in the control of Telomeric Position Effect (TPE), a gene silencing
mechanism mediated by the TAS repeats.
51

A genome-wide RNAi screen for Neuroblast cell cycle exit in
Drosophila
Catarina C. Homem and Juergen A. Knoblich
Institute of Molecular Biotechnology of the Austrian Academy of
Sciences (IMBA)
During development, Drosophila neural stem cells, the Neuroblasts
(Nbs), divide asymmetrically to self-renew and to generate a
differentiated Ganglion Mother Cell (GMC). The GMC divides once
more to generate two post-mitotic neurons or glia. Drosophila Nbs
undergo multiple rounds of divisions generating hundreds of neurons,
which make up the nervous system of the fly. Drosophila Nbs divide
throughout development, but stop dividing and disappear just before
entering adult stages. To maintain the correct number and type of
neurons, it is essential to precisely regulate the time at which
neurogenesis ceases. Although several factors influencing neural
proliferation have been identified, the underlying molecular
mechanism scheduling the end of progenitor divisions remains
enigmatic. Tight control of the number of self-renewing and
differentiating daughter cells is also crucial to prevent uncontrolled
proliferation and tumor formation.
To identify novel genes regulating stem cell proliferation and division
termination we are carrying out a genome wide in vivo RNAi screen
using Nbs as a model. In our assay we co-express Luciferase and
RNA hairpins specifically in a subset of Nbs and their respective
progeny. We then measure luminescence amounts in the adult head
to assess Nb number and lineage sizes.
Up to now we have screened a total of 7500 RNAi lines targeting
53.6% of the protein coding genes in the Drosophila genome. We
have identified significant increases in luminescence in 6.8% of the
RNAi lines screened, corresponding to 402 genes. In this group, we
expect to find several classes of genes involved in the regulation of
Nb size, life-span and proliferation.
Consistent with our hypothesis, among these we find Brat, Numb and
Miranda, known to cause Nb number increase and tumor formation.
In a secondary analysis by immunofluorescence we have, so far,
identified three previously uncharacterized potential tumor
suppressors and two new genes potentially involved in Nb life-span
regulation.
52

A bit of T cell immunology, or playing a different type of CD
Bruno Silva-Santos
Insituto de Medicina Molecular, Lisboa
For an immunologist, fellow non-immunologist scientists must be
often treated as “lay audience” due to heavy jargon that includes
many CD numbers (CD3, CD127…) and an increasing amount of Th
subsets (from Th1 to Th22 or more…). While keeping that in mind, I
will try to summarize the research developed in my lab over the past
4 years. It will be a rapid journey on T cell differentiation and T cell
responses to infectious microrganisms and tumours. Instead of the
most common CD4+ or CD8+ lymphocytes, we will mostly discuss
the unconventional gamma-delta T cell subset. I leave you the three
take home messages: my favourite CD number is 27 because CD27
plays key roles in thymic development and peripheral expansion of
gamma-delta T cells; ULBP1 is the critical tumour antigen for gammadelta
T cell recognition of haematological tumours; and we are now
studying the molecular (epigenetic-to-transcriptional-to-posttranscriptional)
mechanisms of Th1 and Th17 differentiation in vivo…
Well, I will try to be clearer during the talk…
53

Quiescent neuroblasts are reactivated via TOR and paracrine Insulin
relays in Drosophila
Rita Sousa-Nunes, Lih Ling Yee and Alex P. Gould
Division of Developmental Neurobiology, Medical Research Council
National Institute for Medical Research, The Ridgeway, Mill Hill,
London NW7 1AA, UK
Many stem, progenitor and cancer cells undergo periods of mitotic
quiescence from which they can be reactivated. The signals triggering
entry into and exit from this reversible dormant state are not well
understood. Multipotent self-renewing progenitors in the developing
Drosophila central nervous system (CNS), termed neuroblasts,
undergo quiescence in a stereotypical spatiotemporal pattern. Entry
into quiescence is regulated by Hox proteins and an internal
neuroblast timer. Exit from quiescence (reactivation) is subject to a
nutritional checkpoint requiring dietary amino acids. Organ co-culture
experiments also implicate an unidentified signal from an
adipose/hepatic-like tissue called fat body. However, it has been
unclear how these CNS-extrinsic nutritional cues are transmitted to
quiescent neuroblasts. Here, we provide in vivo evidence that
neuroblasts are able to sense amino acids indirectly, via a tissue-totissue
relay: fat body and CNS glia, neurons and neuroblasts. We find
that a signal secreted by the fat body, dependent upon amino-acid
and Target-of-Rapamycin (TOR) signalling, is required for neuroblasts
to exit quiescence. We also demonstrate that Insulin-like Peptides
(Ilps), produced by CNS glia/neurons in a nutrient-dependent
manner, are necessary and sufficient for timely neuroblast
reactivation but do not regulate overall larval size. Conversely,
systemic Ilps secreted into the hemolymph by median neurosecretory
cells (mNSCs) regulate organismal size but do not reactivate
neuroblasts. Drosophila thus contains two segregated Ilp pools, one
regulates neuroblast proliferation within the CNS and the other
controls tissue growth systemically. Together, our findings support a
tissue relay model in which amino-acid sensing by the fat body
stimulates Insulin signalling from glia/neurons, triggering
Phosphatidylinositol 3-Kinase (PI3K)/TOR activation in neuroblasts, in
turn leading to cell cycle re-entry. This mechanism links syst
emic and tissue-restricted signals to neural proliferation and, more
generally, highlights that dietary nutrients and remote organs are key
regulators of transitions in stem-cell behaviour.
54

CISA Project – Developing a Health Research Center in Angola
Susana Vaz Nery
CISA Project
The “CISA Project”, a partnership between the Angolan Ministry of
Health, the Bengo Provincial Government, the Portuguese
Development Aid Institute and the Calouste Gulbenkian Foundation,
aims to develop a Health Research Centre in Angola (CISA), located
in the city of Caxito, Dande´s municipality, Bengo´s Province.
The “CISA Project” promotes epidemiological and clinical research on
the most prevalent or relevant diseases in Angola, such as malaria,
tuberculosis and neglected tropical diseases (including
schistosomiasis and intestinal parasitoses).
The Calouste Gulbenkian Foundation leads the CISA Project
management, whereas in Caxito research activity is implemented by
a team of more than 60 people.
In 2009 a Demographic Surveillance System (DSS) has been
established, with the aim of characterizing the size and dynamics of
Dande´s population and its geographical distribution, thus providing
background information for epidemiological studies. The initial census
of the DSS-Dande was completed in April 2010, covering a population
of more than 60,000 residents.
A baseline prevalence survey on malaria, schistosomiasis, intestinal
parasitoses, anaemia and malnutrition in the DSS study area is being
completed, and it will sample approximately 3000 people. In parallel
to this study an ethnobotanic baseline survey was also implemented.
Other scientific activities initiated in 2010 include: 1) a pilot to
establish a verbal autopsy system in order to determine the main
causes of deaths that happen in the community; and 2) a pediatrics
clinical surveillance system that will provide a platform for clinical
research studies besides improving the quality of documentation and
care at the hospital.
55

Adaptation in Escherichia coli
Isabel Gordo
Instituto Gulbenkian de Ciencia
Adaptation involves changes in the genotype, which will then have
phenotypic consequences that increase the ability of survival and
reproduction of organisms in a given environment. The problem is
that organisms adapt through mutations that are random with
respect to their needs. In order to understand the process of
adaptation we are performing experimental evolution. We are
studying the rate and effects of adaptive mutations in populations of
Escherichia coli, adapting to different environments. We interpret our
results on the fitness effects of new adaptive mutations in the light of
Fisher's geometrical model and are developing theoretical extensions
of this model.
56

Membrane trafficking in synaptic development
Rita O. Teodoro and Thomas L. Schwarz
F.M. Kirby Neurobiology Center, Children’s Hospital Boston and
Department of Neurobiology, Harvard Medical School
Neurons are specialized cells that can extend over great distances,
enabling the complex networking of the nervous system. Like all
other eukaryotic cells, it is essential for neurons to have tight spatial
control of exocytosis in order to form and maintain specialized
domains within the cell. Membrane trafficking is emerging as a key
aspect of neuronal development and function. The exocyst is a
protein complex linked to membrane traffic and one way it is
regulated is by its interaction with several small GTPases, including
RalA. Although not required for the exocytosis of synaptic vesicles,
the exocyst may be important in other aspects of synapse growth and
plasticity through its involvement in and regulation of the tethering,
docking and fusion of post-Golgi vesicles with the plasma membrane.
I will present evidence of a signaling pathway by which synaptic
activation can recruit the exocyst complex to postsynaptic zones and
promote growth of postsynaptic membranes. For a more complete
understanding of the molecular pathways by which the exocyst
contributes to synaptic development, we did a forward genetic screen
to identify genes that interact with the exocyst member Sec5.
Altogether, our studies identified a novel pathway by which synaptic
activity, with its concomitant influx of Ca2+, can contribute to the
development of the post-synapse and, potentially, to a form of
activity-dependent plasticity involving membrane growth. Finally, we
hope to further understand and dissect new pathways that contribute
to synaptic development and plasticity by uncovering new genes that
genetically interact with exocyst.
57

Host-Microorganism Interactions
Luis Teixeira
Instituto Gulbenkian de Ciência
Multicellular organisms and microorganisms are continuously
interacting. Many of these interactions are mutually beneficial.
However, multicellular organisms have to actively thwart invasion by
opportunistic or overtly pathogenic microbes. We are studying the
interaction of the model organism Drosophila melanogaster with
different microorganisms, in particular intracellular ones.
D. melanogaster has been successfully used as a model system to
study innate immunity against many pathogens. Recently it has been
shown that there are innate immunity pathways against viruses
conserved between insects and mammals. We are investigating
mechanisms of resistance to viruses in the fruit fly.
Interestingly, we have found that the intracellular bacteria Wolbachia
confer resistance to RNA viruses in D. melanogaster. We want to
understand the molecular basis of this induced resistance.
Finally, we are interested in the interplay between Drosophila and
Wolbachia itself. These endosymbionts are one of the most
widespread intracellular bacteria in the world but little is known, at
the molecular level, on how the hosts control Wolbachia or Wolbachia
manipulate the hosts.
58

Into the molecular nature of the telomeric checkpoint Inhibition
Tiago Carneiro and Miguel Godinho Ferreira
Instituto Gulbenkian de Ciência, Oeiras, Portugal
Telomeres, the natural chromosome ends of eukaryotes, have unique
properties that distinguish them from damage-induced DNA double
strand breaks (DSB). Whereas a single DSB can halt cell cycle
progression and activate repair processes, telomeres prohibit
checkpoint activation and DNA repair. Nevertheless, several
components involved in DNA damage response (DDR) are critical for
end protection and must be controlled at telomeres to prevent
checkpoints.
We recently showed that telomeres constitute a chromatin-privileged
region capable of stably mounting DDRs without blocking the cell
cycle.
Rather than preventing recruitment and activation of Rad3ATR
kinase, telomeres exclude the mediator Crb253BP1 protein thus
preventing DDR amplification and full checkpoint response. In
contrast to the rest of the genome, H4K20me2 (but not γH2A) is
absent from telomeres. Lack of H4K20me2 from chromosome-ends
cannot be explained by absence of Set9 methyltransferase. Not only
telomeres exhibit Set9-dependent mono and trimethylated forms of
H4K20 but also artificial recruitment of Set9 to chromosome-ends
does not restore a full checkpoint response.
We are currently investigating how telomere heterochromatin
excludes H4K20me2. We are devising two complementary assays: 1-
we hope to determine sufficiency of telomere proteins to prevent
checkpoints by tethering them adjacent to a HO-induced double
strand break; 2- conversely, we are analysing internal telomere
sequences to study whether these are sufficient to modify the
surrounding chromatin and exclude epigenetic markers required for
checkpoint response.
59

Fundraising for science-New approaches for research funding in
Portugal
Maria João Leão
Instituto Gulbenkian de Ciencia, Lisboa, Portugal
IGC has recently begun to develop fundraising initiatives that involve
the scientific community, the private sector and the general public.
This project aims to establish alternative means of financial resources
for research in Portugal and to contribute to a closer interaction
between Research Institutions and the Portuguese society.
60

Plenary Session
Jonathan Howard
Univeristy of Colgne, Germany
“How to build a perfect institute”
62

ABSTRACTS FOR ORAL PRESENTATIONS
Tuesday 28 December
SESSION FOUR
64

The force of the pulsating collective
Antonio Jacinto
Insituto de Medicina Molecular, Lisboa
We still do not know very much about coordination of forces
generated by cells during morphogenesis. It amazes me how an
entity with the complexity, fluidity, and softness of a cell can work
with its neighbours to drive dynamic but extremely robust tissue
shape changes that occur during embryogenesis or regeneration. I
will tour recent results from our group that highlight the importance
of non-monotonic cell behaviours; pulses of forces generated by the
actomyosin network seem to be one of the crucial strategies that cells
use to work together to produce from.
66

Overview of the Cell Death Regulaion Laboratory
Luis Miguel Martins
Medical Research Council Toxicology, Leicester University, UK
Tissue homeostasis relies on the tightly controlled removal of
superfluous, damaged and ectopic cells, normally through apoptosis.
Whereas an aberrant resistance to apoptosis participates in the
development of neoplasias, excessive cell death, through apoptosis or
necrosis, contributes to acute organ failure as well as to chronic
diseases involving the loss of post-mitotic cells.
The focus of our research is to better understand the control of
cellular death processes in normal cells as well as in pathological
states where alterations of these processes have occurred.
Our aim is to achieve a better understanding of the regulatory
mechanisms controlling cell death. Understanding the basic biology of
these processes can provide insights that might lead to targeted
treatment solutions that will act on pathways and alter disease at its
cause resulting in a better type of medicine.
67

Establishment of asymmetries in the early vertebrate embryo
José António Belo
(1) Regenerative Medicine Program, Departamento de Ciências
Biomédicas e Medicina, Universidade do Algarve, Portugal
(2) IBB-Institute for Biotechnology and Bioengineering, Centro de
Biomedicina Molecular e Estrutural, Universidade do Algarve, Portugal
(3) Instituto Gulbenkian de Ciência, Oeiras, Portugal
One fundamental aspect of vertebrate embryonic development is the
formation of the body plan. For this process, asymmetries have to be
generated during early stages of development along the three main
body axes: Anterior-Posterior, Dorso-Ventral and Left-Right. We have
been studying the role of a novel class of molecules, the
Cerberus/Dan gene family. These are dedicated secreted antagonists
of three major signaling pathways: Nodal, BMP and Wnt. Our studies
contribute to the current view that the fine tuning of signaling is
controlled by a set of inhibitory molecules rather than by activators.
In this context, the Cerberus-like molecules emerge as key players in
the regulation and generation of asymmetries in the early vertebrate
embryo. Recent studies conducted in our lab unraveled the role of
these molecules during organogenesis as well.
68

Development on time: the embryonic molecular clock
Isabel Palmeirim
Medical and Biomedical Sciences Department, Bio-regenerative
Medicine Program, IBB-Institute for Biotechnology and
Bioengineering, Centre for Molecular and Structural Biomedicine,
University of Algarve, Campus de Gambelas, 8005-139
Alongside the three dimensions in which an embryo undergoes
growth and differentiation, one can consider TIME as being a fourth
dimension, which must experience equally tight regulatory
mechanisms in order to assure that each tissue/structure/organ is
formed in the right place, at the right time. Time control is
particularly evident during the process by which the vertebrate
presomitic mesoderm is progressively segmented into repeated units,
named somites. Somites arise in a highly coordinated way both in
space and time and, later on, give rise to vertebrae, intervertebral
discs, ribs and skeletal muscles. In 1997, the first molecular evidence
for the existence of an intrinsic cellular molecular clock underlying the
rhythm of somitogenesis has been provided. We currently know that
this clock operates in all vertebrate groups studied, in more than one
embryonic tissue, with different time-periods and an increasing
number of genes belonging to the Notch, FgF and Wnt
signalling pathways are being implicated in this clocked mechanism.
Recently, our work brings forth new results, evidencing the role of
Shh signaling pathway in pacing molecular clock gene expression and
timing morphological somite formation. We believe that a deeper
understanding of the role and regulation of this embryonic
mechanism is of crucial importance, since temporal control of
biological processes is a transversal issue in all life sciences research
fields.
69

NK cells in Preeclampsia
Ana Sofia Cerdeira, Hernan D. Kopcow, Zaheed Husain, Agnes Lo,
Caroline Royle, Ravi I. Thadhani, Isaac Stillman, Vikas P. Sukhatme
and S. Ananth Karumanchi
Gulbenkian Programme for Advanced Medical Education
Beth Israel Deaconess Medical Center/Harvard Medical School
Serviço de Ginecologia e Obstetricia Centro Hospitalar do Porto
Preeclampsia is a pregnancy disease characterized by new onset
hypertension and proteinuria. It is a leading cause of maternal and
perinatal morbidity and mortality complicating 3-8% pregnancies.
Recent evidence suggests that Natural Killer (NK) cell signaling at the
maternal-fetal interface is an important regulator of maternal spiral
artery remodeling. This process is impaired in preeclampsia and leads
to increased expression of placental soluble factors such as sFlt1 that
induce the systemic manifestations of the disease. NK cells are
abundant at the maternal decidua (dNK) during the first trimester of
pregnancy, and show severely reduced cytotoxicity and secretion of
proangiogenic factors like VEGF. In contrast, peripheral blood NK cells
(pNK) are cytotoxic and non-angiogenic. We hypothesized that a
combination of hypoxia and immunosuppressive cytokines like TGF-
β1, known to be present in the placenta, may turn pNKs into dNKlike
proangiogenic cells with reduced cytotoxicity.
Like dNKs, pNKs cultured under hypoxia secreted VEGF and induced
tube formation by HUVECs. Under hypoxia in the presence of TGF-β1
pNKs showed reduced cytotoxicity. Since placenta and tumor
microenvironment share many features, we characterized tumor
infiltrating NK cells (tiNK) to better understand this phenomenon.
tiNK cells isolated from kidney tumors, like dNKs, show reduced
cytotoxicity and secrete VEGF. Moreover, pNK cells exposed to
hypoxia and TGF-β1 support tumor growth in a mouse model of
cancer. The hypoxic and immunosuppressive microenvironment of
both tumor and placenta may co-opt NK cells transforming them into
poorly cytotoxic cells that secrete VEGF. In vitro conversion of pNKs
into angiogenic dNK-like cells may have potential applications in the
development of novel therapeutic approaches for preeclampsia.
70

The heterogenous behavior of human breast cancer
Sofia Braga (1,2,3) and José B. Pereira-Leal (1)
(1) Instituto Gulbenkian de Ciência - Oeiras, Portugal
(2) Instituto Português de Oncologia - Lisboa, Portugal
(3) Programa Gulbenkian de Formação Médica Avançada
BC is a heterogenous disease. There are three subtypes of BC,
defined by immunohistochemical detection of specific proteins, with
prognostic and therapeutic implications: ER positive BC, half of which
also express the PgR, Her2 positive BC, and Triple negative BC. ER+
BC is generally considered easier to manage clinically because of the
sensitivity to anti-estrogenic therapy, Her2+ is treated with anti-Her2
therapy but has worse prognosis and TNBC has no validated targeted
therapy having the worse prognosis. Clinicians observe heterogeneity
in the outcome of BC: not all ER+ BC are cured, some Her2+ BC
relapse during therapy and not all TNBC have a uniformly dismal
prognosis. Our goal is to dissect the molecular basis of this
heterogeneity. We already compiled and analyzed publicly available
expression data (microarrays) from 3500 human tumors with
outcome annotation. We performed subtype selection by standard
immunohistochemical annotation and by using a computational
algorithm. We used classical supervised methods for discovering
genes and pathways that are differentially expressed in tumours with
differing outcome. We have generated a set of potential prognostic
markers for each subtype of BC that we are now further
investigating. The next step in this analysis is a prospective
validation of the prognostic value of these genes and pathways in
collaboration with pathologists at public hospitals in Lisbon.
71

Different tasks, different tradeoffs: speed and accuracy in odor
detection vs. odor categorization
Maria I. Vicente, André G. Mendonça, Zachary F. Mainen
Systems Neuroscience Lab, Champalimaud Neuroscience Programme
Speed-accuracy tradeoffs (SATs) are well known in cognitive and
perceptual decision-making, and can involve large (>50%) changes
in processing time. However, in some tasks, such as odor mixture
categorization, much smaller SATs (order 10%) are observed. Why
some tasks benefit more from temporal processing than others is not
well understood.
We hypothesize that the degree of SAT depends on the relative
contribution of sensory and non-sensory noise. To test this idea, we
compared SAT in the odor mixture categorization task to that in a
new two alternative choice odor detection task, using the same odors
diluted up to 1,000-fold. The same rats performed both tasks and all
other task variables were held constant. We envisaged that stimulus
noise would dominate the detection but not the categorization task.
As predicted, we found that SAT (difference in reaction time, RT, from
the easiest to the hardest problem) was much larger in the odor
detection task (53% increase in RT as accuracy decreased from 97%
to 62%) than in the mixture categorization task (14% increase for a
similar change in accuracy). These data demonstrate that sensory
integration and the amount of SAT are problem-specific and suggest
that the locus of performance-limiting noise is a critical variable.
72

Chromatin dynamics in motor skill learning
Galvão-Ferreira P. and Costa R. M.
Champalimaud Neuroscience Program at Instituto Gulbenkian de
Ciencia, Rua da Quinta Grande 6, 2781-901 Oeiras, Portugal
Motor skills can last a lifetime. In previous work, we have shown that
motor skill learning involves changes in synaptic plasticity in the
striatum which evolve with training. In addition, we observed
differential involvement of striatal output circuits (namely,
striatonigral and striatopallidal circuits) in skill authomatization. In
recent years, both DNA methylation and post-translational
modifications of nuclear proteins have been implicated in
physiologically normal mechanisms (such as synaptic plasticity and
memory formation) as well as in neuropathological contexts. Yet, the
involvement of these epigenetic mechanisms in motor skill learning
has not been investigated.
To test for the involvement of these biochemical mechanisms in skill
learning, we trained young and old mice (2 and 8 months old,
respectively) in a lever-pressing task, in which subjects had to press
the lever at up to 8Hz. Next, using DRD1-GFP and DRD2-GFP reporter
mice to distinguish between D1-expressing and D2-expressing
medium spiny (respectively, striatonigral and striatopallidal) neurons,
we inquired whether epigenetic changes could be found in these two
circuits through immunohistochemistry and Western blotting.
Currently, we are using those same transgenic mouse lines in an
effort to extract and purify striatonigral and striatopallidal neurons
through fluorescent activated cell sorting (FACS). Using these varied
approaches, we intend to investigate if the long-lasting plasticity
observed throughout skill learning involves defined epigenetic
mechanisms in the striatonigral and striatopallidal output circuits.
73

Plenary Session
Maria Leptin, EMBO, Germany
“My Life in Science”
74

Alexandra Borges
Participants List
Instituto de Investigaciones Biomédicas Alberto Sols, Spain
borgalexandra@gmail.com
Alexandra Capela
StemCells Inc, USA
alexandra.capela@stemcellsinc.com
Alexandra Santos
King's College London, Hospitais da Universidade de Coimbra, UK
alexandrafigueirasantos@gmail.com
Alexandre Raposo
Instituto Gulbenkian de Ciência
araposo@igc.gulbenkian.pt
Ana Borges
Instituto Gulbenkian de Ciência, PT
aborges@igc.gulbenkian.pt
Ana Campilho
IBMC, PT
anacampilho@ibmc.up.pt
76

Ana Mateus
Cambridge University, UK
amam3@cam.ac.uk
Ana Matos
Instituto Gulbenkian de Ciência
anamatos.ct.surgery@gmail.com
Ana Sousa
Hospital de Santa Maria, PT
anabertasousa@gmail.com
Ana Sofia Cerdeira
Beth Israel Deconess Medical Center, USA
ana.cerdeira@gmail.com
Ana Teresa Tavares
CIISA-FMV, PT
atavares@igc.gulbenkian.pt
Ana Sanchez
ITQB, PT
asanchez@itqb.unl.pt
77

António Coutinho
Instituto Gulbenkian de Ciência
coutinho@igc.gulbenkian.pt
António Jacinto
Inst. de Medicina Molecular, Fac. de Medicina da Univ. de Lisboa, PT
ajacinto@fm.ul.pt
Carla Martins
Cancer Research, UK
Carla.Martins@cancer.org.uk
Catarina Homem
Inst. of Molecular Biotech. of the Austrian Ac. of Sciences (IMBA), AUSTRIA
catarina.homem@imba.oeaw.ac.at
Cátia Proença
Gulbenkian/Weill Cornell, USA
cproenca@igc.gulbenkian.pt
César Evaristo
University of Chicago, USA
cesar.evaristo@gmail.com
78

Cláudio Gomes
ITQB
gomes@itqb.unl.pt
David Cristina
Instituto Gulbenkian da Ciência, PT
davidc@igc.gulbenkian.pt
Dinis Calado
Harvard Medical School, USA
calado@idi.harvard.edu
Diogo Castro
Instituto Gulbenkian de Ciência, PT
dscastro@igc.gulbenkian.pt
Diogo Lucena
Fundação Calouste Gulbenkian
dlucena@gulbenkian.pt
79

Diogo Pimentel
DPAG- Oxford University, UK
d.o.pimentel@gmail.com
Eduardo Silva
Wyss Institute/ Harvard University, USA
easilva@wyss.harvard.edu
Filipa Barbosa
Insituto Gulbenkian de Ciência
fbarbosa@igc.gulbenkian.pt
Filipe Martins
Dana-Farber Cancer Institute, UK
filipecorreiamartins@gmail.com
Francisco Dionísio
Centro de Biologia Ambiental, Fac. de Ciências Univ. de Lisboa, PT
francisco.dionisio@gmail.com
Frederico Regateiro
University of Oxford, UK
regateiro@gmail.com
80

Guilherme Neves
Molecular Neurobiology Div., MRC- Nat. Inst. for Medical Research, UK
gneves@nimr.mrc.ac.uk
Greta Martins
Instituto Gulbenkian de Ciência
gmartins@igc.gulbenkian.pt
Helena Soares
Pasteur Institute, FR
helena.soares@pasteur.fr
Inês Pires da Silva
New York University, USA
inespiresilva@gmail.com
Isabel Campos
IMM, PT
icampos@fm.ul.pt
Isabel Duarte
Instituto Gulbenkian de Ciência, PT
iduarte@igc.gulbenkian.pt
81

Isabel Gordo
Instituto Gulbenkian de Ciência, PT
igordo@igc.gulbenkian.pt
Isabel Palmeirim
Instituto Gulbenkian de Ciência, PT
palmeiri@igc.gulbenkian.pt
Joana Barros
Associação viver a ciência, PT
jbarros@viveraciencia.org
Joana Sá
Harvard University, USA
mjsa@fas.harvard.edu
Joana Santos
Instituto Gulbenkian de Ciência, PT
joanars@igc.gulbenkian.pt
Jonathan Howard
Institute for Genetics
82

José A. Belo
Dep. de Ciências Biomédicas e Medicina, Univ. Do Algarve, PT
jbelo@ualg.pt
José Antão
Massachusetts General Hospital, USA
antao@molbio.mgh.harvard.edu
José Leal
Instituto Gulbenkian de Ciência, PT
jleal@igc.gulbenkian.pt
Leonor Parreira
Universidade de Lisboa, Faculdade de Medicina/ IGC
lparreir@igc.gulbenkian.pt
Lília Perfeito
University of Cologne, GER
lilia.perfeito@gmail.com
L. Miguel Martins
MRC Toxicology Unit, UK
martins.lmiguel@gmail.com
83

Luís Graça
Instituto de Medicina Molecular, PT
lgraca@fm.ul.pt
Luis Soares
Harvard Medical School, UK
luis_soares@hms.harvard.edu
Luis Teixeira
Instituto Gulbenkian de Ciência, PT
lteixeira@igc.gulbenkian.pt
Manuel Batista
Skirball Institute NYU, USA
mfscb2@googlemail.com
Manuel Rebelo
Instituto Gulbenkian de Ciência, PT
mrebelo@igc.gulbenkian.pt
Margarida Trindade
IMM, PT
mtrindade@fm.ul.pt
84

Maria Inês Vicente
Champalimaud Neuroscience Program at IGC, PT
mivicente@igc.gulbenkian.pt
Maria Leptin
EMBO, Germany
Maria Luisa Vasconcelos
CNP at Instituto Gulbenkian de Ciência, PT
vasconcelos@igc.gulbenkian.pt
Maria João Leão
Instituto Gulbenkian de Ciência, PT
mjleao@igc.gulbenkian.pt
Marta Nunes
University of the Witwatersrand, SOUTH AFRICA
mnunes00@yahoo.com
Miguel Gaspar
mangerico@gmail.com
85

Miguel God. Ferreira
Instituto Gulbenkian de Ciência, PT
mgferreira@igc.gulbenkian.pt
Miguel Remondes
MIT, USA
remondes@mit.edu
Mónica Dias
Instituto Gulbenkian da Ciência, PT
mdias@igc.gulbenkian.pt
Mónica Sousa
IBMC, PT
msousa@ibmc.up.pt
Nuno Costa
Institute of Neuroinformatics, Switzerland
ndacosta@ini.phys.ethz.ch
Ofélia P. De Carvalho
University of Cambridge, UK
omc20@cam.ac.uk
86

Paula Duque
Instituto Gulbenkian de Ciência, PT
duquep@igc.gulbenkian.pt
Paulo Ribeiro
London Research Institute, Cancer Research, UK
paulo.ribeiro@cancer.org.uk
Pedro Batista
Wake Forest Institute for Regenerative Medicine, USA
pbaptist@wfubmc.edu
Pedro Brites
IBMC, PT
pedro.brites@ibmc.up.pt
Pedro Domingos
ITQB, PT
domingp@itqb.unl.pt
Pedro Ferreira
Champalimaud Neuroscience Program at IGC, PT
pferreira@igc.gulbenkian.pt
87

Perpétua Pinto-do-Ó
INEB-Instituto Nacional de Engenharia Biomédica, PT
perpetua@ineb.up.pt
Ricardo Almeida
University of Edinburgh, UK
r.almeida@ed.ac.uk
Ricardo Costa
Centro Nacional de Investigaciones Cardiovasculares – CNIC, SP
ricardo.costa@cnic.es
Ricardo Neto Silva
Champalimaud Neuroscience Programme at Instituto Gulbenkian de Ciência, PT
ricmigneto@gmail.com
Rita Sousa Nunes
National Institute for Medical Research (NIMR), UK
rsousa@nimr.mrc.ac.uk
Rita Tavares
University of California, San Francisco, USA
ritamtavares@gmail.com
88

Rita Teodoro
Harvard Medical School, USA
roteodoro@yahoo.com
Rui Goncalo Martinho
Instituto Gulbenkian de Ciência, PT
rmartinho@igc.gulbenkian.pt
Rui Peixoto
Instituto Gulbenkian de Ciência
rmartinho@igc.gulbenkian.pt
Severina Moreira
University of Bristol, UK
severina_moreira@hotmail.com
Sheila Vidal
Instituto Gulbenkian de Ciência, PT
svidal@igc.gulbenkian.pt
Silvia Costa
ISPA, PT
silviasantoscosta@gmail.com
89

Sofia Araujo
IBMB-CSIC, SPAIN
sofia.araujo@irbbarcelona.org
Sofia Braga
Instituto Gulbenkian de Ciência, PT
sbraga@igc.gulbenkian.pt
Susana Vaz Nery
CISA PROJECT, ANGOLA
susana.nery@gmail.com
Tiago Botelho
IBMB-CSIC, SP
tobcri@ibmb.csic.es
Tiago Carneiro
Instituto Gulbenkian de Ciência, PT
tcarneir@igc.gulbenkian.pt
Tiago Carvalho
LabOrders, PT
tiago@laborders.com
90

Ulla-Maj Fiuza
No instituition affiliation
umfiuza@igc.gulbenkian.pt
Vasco Barreto
Instituto Gulbenkian de Ciência, PT
vbarreto@igc.gulbenkian.pt
Vanessa Luis
Instituto de Medicina Molecular, PT
vluis@fm.ul.pt
Vera Teixeira
Instituto de Medicina Molecular, PT
teixeira@igc.gulbenkian.pt
91