Current Honours Projects - Faculty of Medicine, Nursing and Health ...

Department of Pharmacology, Honours 2013 

Honours in Pharmacology 

2013 

www.monash.edu 

Copyright © Monash University 2012. All rights reserved. Except as provided in the Copyright Act 1968, this 

work may not be reproduced in any form without the written permission of the host Faculty and 

School/Department.


Welcome to the Pharmacology Department! 

The Honours year represents a new adventure, very different to your undergraduate 

experience, in which you will have the opportunity to undertake a research project, 

communicate your science to colleagues and peers and learn to critically evaluate scientific 

concepts and literature. 

Your supervisor(s) will be there to guide and advise you along this research journey. 

At the very least, you are expected to bring with you the following skills set, in no particular 

order: 

-enthusiasm 

-an enquiring mind 

-respect & humility 

-determination & persistence 

-a sense of humour 

-a collegial spirit 

-patience 

This booklet provides information about the research projects on offer in the Department of 

Pharmacology and we encourage you to identify the areas of research in which you are most 

interested, contact potential supervisors and discuss the projects with them. 

The course convenors, A/Prof Grant Drummond and Dr Barb Kemp-Harper, can advise on 

projects, guide you through the application process and help with any queries you may have. 

We look forward to welcoming you the Department of Pharmacology in 2013 and wish you 

all the best for a rewarding and exciting year of research. 

Good luck! 

Professor Robert Widdop 

Head, Department of Pharmacology 



School/Department.


Department of Pharmacology Honours Convenors 

Dr Barb Kemp-Harper A/Prof Grant Drummond 

Email: Barbara.Kemp@monash.edu Email: Grant.Drummond@monash.edu 

Phone: 9905 4674 Phone: 9905 4869 

Pharmacology Honours: Pre-requisites 

BSc / BBNSc / BBiotech BMS 

Pre-requisites A Distinction average (>70) 

in 24 points at 3 rd year level, 

including at least 18 points 

in 3 rd year PHA units 

A Distinction average (>70) 

in 24 points at 3 rd year 

level, including 12 points in 

3 rd year core BMS units 

and 12 points in other 3 rd 

year units 

Application closing date 19 th November, 2012 16 th November, 2012 

Application form www.sci.monash.edu.au/undergra 

d/honours/apply.html 

www.med.monash.edu/biomed/ 

honours/ 

Commencement date 25 th February, 2013 25 th February, 2013 



School/Department.


Honour Projects 2013 – On campus projects 

LABS / SUPERVISOR(S) 

Clinical & Experimental Toxicology 

Group 

Andis Graudins 

Education-based projects 

Eva Patak, Liz Davis 

Fibrosis Group 

Chrishan Samuel, Simon Royce 

Integrative Cardiovascular 

Pharmacology Group 

Tracey Gaspari, Rob Widdop 

Claudia McCarthy, Antony Vinh, Rob 

Widdop 

Antony Vinh, Rob Widdop 

Neuropharmacology Group 

Richard Loiacono, Siew Yeen Chai 

(Physiology) 

Vascular Biology & 

Immunopharmacology Group 

Brad Broughton, Chris Sobey 

Grant Drummond, Rebecca Lim 

Barb Kemp-Harper 

Barb Kemp-Harper, Grant Drummond 

Helena Kim, Chris Sobey 

Alyson Miller 

Stavros Selemidis 

PROJECT TITLE 

• An assessment of fructose-1,6-diphosphate alone and in 

concert with other inotropic agents on inotrope sparing 

in the treatment of beta-receptor antagonist poisoning 

• Evaluation of web-based pharmacology resources 

• Investigating novel anti-fibrotic therapies 

• Cardio-protective effects of incretin hormones: potential 

beyond glycaemic control? 

• Mechanisms of angiotensin type-2 receptor-mediated 

neuroprotection during stroke 

• Role of T cells during the development of high blood 

pressure 

• Modulation of neuronal oxytocin – effects on social 

behaviour 

• Are the adverse effects of hormone replacement therapy 

on stroke outcome mediated by G protein-coupled 

estrogen receptor signalling? 

• Stem cell therapy for the treatment of high blood 

pressure 

• Exploring the vasoprotective actions of novel HNO 

donors 

• Exploring novel therapeutic strategies to prevent 

atherosclerotic plaque rupture 

• Exploring the role of endothelial NF-KB on ischemic 

stroke outcome 

• Investigating mechanisms linking hypertension and 

cognitive impairment 

• Novel pharmacological targets for Influenza A virus 

induced lung inflammation 

• Understanding the biology of reactive oxygen species in 

cancer 



School/Department.


Honour Projects 2013 – Off campus projects 

LABS/SUPERVISOR(S) 

Australian Centre for Blood 

Diseases 

Justin Hamilton, Grant Drummond 

Baker IDI Heart & Diabetes Institute 

Jaye Chin-Dusting, Jennifer Irvine, 

Rob Widdop 

Geoff Head, Pamela Davern 

Geoff Head, Pamela Davern, 

Sandra Burke 

Geoff Head, Pamela Davern, Joon Lim 

Rebecca Ritchie, Barb Kemp-Harper 

Rebecca Ritchie 

Drug Discovery Biology Theme 

Monash Institute of Pharmaceutical 

Sciences 

Michelle Halls, Roger Summers 

Rob Lane, Arthur Christopoulos 

Katie Leach, Laura Lopez, Arthur 

Christopoulos 

Katie Leach, Arthur Christopoulos 

Patricia Rueda, Arthur Christopoulos, 

Chris Langmead 

Celine Valant, Lauren May 

Celine Valant, Meritxel Canals 

Celine Valant, Arthur Christopoulos 

PROJECT TITLE 

• Defining the function of human platelet thrombin 

preceptor, PAR4, in arterial thrombosis 

• What turns you on? Using a novel RNAi-based 

approach to determine platelet activation signals in 

thrombosis 

• The role of AT2R in macrophage polarisation 

• Effects of positive allosteric modulator of GABAA 

receptors on hypertension and stress 

• Role of brain pathways in chronic stress 

• Which parts of the brain are affected in obesity related 

hypertension? 

• Nitroxyl, a relative of NO, is a naturally-occurring 

cardioprotective molecule 

• New strategies to rescue diabetes induced cardiac 

dysfunction 

• Targeting anti-inflammatory protein annexin-A1 for 

protection from myocardial infarction (heart attack) 

• Compartmentalisation of relaxin-stimulated signalling in 

single cells 

• Molecular mechanisms of a novel allosteric modulator of 

the D2-like dopamine receptors 

• Probing an allosteric binding site in the human CASR 

• Allosteric pharmacoregulation of a headless CASR 

• Investigation of muscarinic M4 receptor function using 

label free impedance technology 

• Investigating the signalling cascade of novel adenosine 

receptor ligands at the adenosine A2B receptor (A2B- 

AR) 

• Investigation of an absolutely conserved pocket at the 

muscarinic acetylcholine M1 receptor (M1 MACHR) 

• Investigation of an absolutely conserved pocket at the 

muscarinic acetylcholine M2 receptor (M2 MACHR) 



School/Department.


AN ASSESSMENT OF FRUCTOSE-1,6-DIPHOSPHATE ALONE 

AND IN CONCERT WITH OTHER INOTROPIC AGENTS ON 

INOTROPE SPARING IN THE TREATMENT OF 

BETA-RECEPTOR ANTAGONIST POISONING 

Supervisors: Professor Andis Graudins 

Location: Clinical and Experimental Toxicology Laboratory 

Department of Pharmacology 

Monash University, Clayton 

Background: 

Heart failure and shock resulting from drug poisoning may be a difficult treatment 

challenge in the clinical setting. Severe poisoning with beta-receptor antagonists such as 

propranolol may result in hypotension (low blood pressure) that may not respond 

adequately to standard treatment measures such as infusion of adrenaline or other 

inotropic therapies. Treatment often results in the administration of multiple medications 

in an attempt to maintain blood pressure. In this stressed state, the myocardium is less 

effective in its ability to produce energy substrates (such as ATP) from glucose and free 

fatty acids. 

Fructose-1-6,diphosphate (FDP) is endogenously produced in human cells as an 

intermediary metabolite of the glycolytic pathway. Its production requires the 

expenditure of 2 molecules of ATP for each molecule of glucose. The administration of 

exogenous FDP improves heart function in a number of models of shock. This is the result 

of improved energy utilization, ATP sparing, improved enzyme activity as well as a number 

of other mechanisms. 

Pilot data from our laboratory has shown that IV administration of FDP to rodents 

poisoned with propranolol resulted in an improvement in blood pressure and survival. This 

was significantly better than that seen in control animals treated either with glucose or 

adrenaline infusion. We do not know whether co-administration of FDP with other 

inotropes such as adrenaline infusion or high-dose insulin infusion results in increased 

improvement in heart function. Also, co-administration of FDP with other agents may 

result in inotrope sparing, thereby reducing the doses of these agents required to improve 

shock. Positive results will help design subsequent clinical trials of FDP in poisoning. 

Project aims: The aim of this honours project is to assess the effects of FDP infusion on 

inotrope sparing, blood pressure, cardiac output and survival in a rodent model of severe 

propranolol poisoning. 

Techniques: Anaesthetised rodents will require surgery to be instrumented to record 

blood pressure, cardiac output and electrocardiographic trace. Intravenous canulae will be 

inserted to infuse propranolol and inotropic treatments. Monitoring and data collection 

using PowerLab hardware and software. 

Contacts: 

Professor Andis Graudins 

Southern Clinical School and Department of Pharmacology, Monash University 

Phone: 95943193, Rm E140; Andis.Graudins@monash.edu 



School/Department.


EVALUATION OF WEB-BASED PHARMACOLOGY RESOURCES 

Supervisors: Dr Elizabeth Davis & Dr Eva Patak 

Location: Pharmacology Education Research Initative 



Background: 

Our group aims to gain a better understanding of factors that influence the engagement 

of students with their learning of pharmacology and the development of resources and 

curricula to support students in their learning, particularly within the science and medical 

courses. This group is currently involved in several ongoing projects and prospective 

students could become involved in aspects of these. 

Project aim: 

The aim of one honours project proposal is to gain a greater understanding of the web 

resources that undergraduate science students use to supplement their learning of 

pharmacology. Student use of information sources will be assessed and popular electronic 

resources will be evaluated for accuracy, reliability and consistency of information. 

Academic and student attitudes towards the use of web resources compared to 

instructor-provided lecture material and recommended textbooks will also be assessed. 

The results of the project will inform the future development of teaching resources. 

This project could also encompass the development of practical / tutorial material – in 

particular, how we can develop practical sessions that address learning outcomes and 

maintain student engagement. 

Techniques: 

In addition to literature searching and reviews to identify what is known of student 

attitudes to learning, research techniques that will be used to complete this project, will 

include the design, development, implementation and analysis of academic/student surveys 

and focus group interviews along with monitoring of the use of lecture recordings and 

Moodle sites. 

Contacts: 

Dr Elizabeth Davis 


Monash University 

Phone: 9905 5755, Rm E123 

Elizabeth.Davis@monash.edu 

Dr Eva Patak 



Phone: 9905 5783, Rm E117 

Eva.Patak@monash.edu 



School/Department.


INVESTIGATING NOVEL ANTI-FIBROTIC THERAPIES 

Supervisors: Dr Chrishan Samuel & Dr Simon Royce 

Location: Fibrosis Laboratory (Rms E101/E102) 



Background: 

Fibrosis is defined as the hardening and/or scarring of various organs including the 

heart, kidney and lung; which usually arises from abnormal wound healing to tissue injury, 

resulting in an excessive deposition of extracellular matrix components, primarily 

collagen. The eventual replacement of normal tissue with scar tissue leads to organ 

stiffness and ultimately, organ failure. Despite a number of available treatments for 

patients with various heart/kidney/lung diseases, patients receiving these therapies still 

progress to end-stage organ failure due to the inability of these treatments to directly 

target the build-up of fibrosis. Hence, novel and more direct anti-fibrotic therapies are 

still required to be established. 

Aims: 

The Fibrosis Lab aims to identify novel anti-fibrotic therapies (relaxin, stem cells, trefoil 

factor 2; and combinations of these) that will more effectively prevent/reverse fibrosis 

progression. Additionally, by understanding the mechanisms of action of these potential 

therapies of future, we aim to delineate new targets that can be utilized to enhance their 

therapeutic potential and abrogation of organ scarring. 

Projects: 

1. Signal transduction studies (in models of heart / kidney / lung disease) 

2. Head-to-head and combination therapy efficacy studies 

3. The influence of ageing and gender on fibrosis 

4. Development of new approaches to target airway remodeling in asthma 

Techniques: 

Depending on the project involved, animal/cell culture models of (heart/kidney/lung 

disease), blood pressure and functional measurements, matrix biology, protein 

biochemistry, molecular biology and/or histological techniques will be utilized. 

Contacts: 

Dr Chrishan Samuel and Dr Simon Royce 



Phone: 9902 0152 / 9905 0913 

chrishan.samuel@monash.edu 

simon.royce@monash.edu 



School/Department.


CARDIO-PROTECTIVE EFFECTS OF INCRETIN HORMONES: 

POTENTIAL BEYOND GLYCAEMIC CONTROL? 

Supervisors: Dr Tracey Gaspari & Prof Robert Widdop 

Location: Cardiovascular Integrative Group 



Collaborator: Dr Anthony Dear 

Deputy Director, Eastern Clinical Research Unit, Monash 

University 

Background: 

Our research focuses on understanding the pathophysiological processes involved in the 

development of cardiovascular disease, one of the major causes of illness and death in 

Australia and worldwide. 

Project aim: 

Two classes of diabetic drugs targeting the glucagon-like peptide-1 (GLP-1) pathway are 

used clinically to treat Type 2 Diabetes Mellitus. These are GLP-1 receptor agonists (e.g., 

Liraglutide) and dipeptidyl peptidase-4 (DPP-4) inhibitors (prolong ½ life of endogenous 

GLP-1). In collaboration with Dr Dear we aim to have projects that investigate the 

vascular and cardio-protective effect of these classes of drugs in a number of 

cardiovascular disease states, including atherosclerosis, vascular injury (neointimal 

hyperplasia) and hypertension. 

Techniques: 

Projects could incorporate a range of methodologies including animal dietary and 

pharmacological treatments, surgical techniques for vascular injury models, 

immunohistochemistry, biochemical measures (western blot, markers of cellular 

proliferation and hypertrophy), histological and morphological analyses (such as fibrosis, 

intimal to medial measurements). 

Contacts: 

Dr Tracey Gaspari and Prof Rob Widdop 



Phone: 9905 4762, Rm E119 

Tracey.Gaspari@monash.edu 

Robert.Widdop@monash.edu 

Figure 1: Representative images of crosssectional 

lesion development in aortic arch of 

ApoE KO mice. Liraglutide treatment shows 

decreased intima (I) development. Intima (I) and 

media (M) areas are identified by dotted line. 



School/Department.


MECHANISMS OF ANGIOTENSIN TYPE-2 RECEPTOR- 

MEDIATED NEUROPROTECTION DURING STROKE 

Supervisors: Dr Claudia McCarthy, Dr Antony Vinh & Prof Rob Widdop 

Location: Integrated Cardiovascular Pharmacology Laboratory 


Monash University, Clayton. 

Background: 

Stroke is the third largest cause of death and the major cause of disability in 

industrialised countries. Every year, approximately 15-20 million people worldwide, will 

suffer a new or recurrent stroke. Despite this, relatively few therapies are available 

for the treatment of stroke. 

The renin angiotensin system (RAS) is a hormonal cascade which has multiple roles 

throughout the body, including the control of the cardiovascular system, renal and 

adrenal function governing fluid and electrolyte balance and arterial pressure. We 

recently demonstrated that a particular subtype of angiotensin receptor, the 

angiotensin II type 2 receptor (AT2 R), is neuroprotective during stroke and has been 

associated with anti-inflammatory effects nerve regeneration following nerve injury. 

Project aims: This study will elucidate mechanics of AT2R mediated neuroprotection 

specifically looking at its interaction with the immune system in determining stroke 

outcome. 

Techniques: An animal model of stroke will be implemented in conjunction with 

behavioural tests to assess stroke severity and any symptomatic relief resulting from 

treatment. Several physiological parameters will be monitored throughout the 

experimental protocol, including blood pressure. Specific organs will be isolated and 

preserved to measure stroke-induced changes using histology. In addition, several 

techniques, including immunohistochemistry and flow cytometry, will identify the 

specific neuronal and immune cells associated with stroke and AT2R function. 

Contacts: 

Dr Claudia McCarthy and Dr Antony Vinh 



Phone: 9905 0095, Rm E119 

Claudia.mccarthy@monash.edu 

Antony.vinh@monash.edu 



School/Department.


ROLE OF T CELLS DURING THE DEVELOPMENT OF HIGH 

BLOOD PRESSURE 

Supervisors: Dr Antony Vinh & Prof Rob Widdop 

Location: Integrative Cardiovascular Pharmacology 



Background: 

Inflammation and immunity continues to be a topical area of hypertension research. The 

adaptive immune system and particularly T cells have recently been implicated in the 

development of hypertension. Interestingly, there are several similarities between 

hypertension and autoimmune disorders, and of particular interest is local infiltration 

of T cells. Like synovial joints of arthritic patients, during experimental hypertension 

there is significantly increased T cell infiltration into non-lymphoid organs such as the 

vasculature, brain and kidneys, which are vital organs that regulate blood pressure. 

While it is hypothesized that these tissue-infiltrating T cells promote local 

inflammation, their precise function remains speculative and undefined. Moreover, the 

timing of the immune response during hypertension is unstudied. 

Project aims: 

The aim of this honours project is to (1) investigate the function of infiltrating T cells 

during hypertension, and through a series time-course experiments, (2) pinpoint the 

sequence of immune-related events leading to activation of the immune system during 

hypertension. This study will identify when and how T cells and the immune system 

contribute to elevating blood pressure, and may lead to novel drug targets to treat this 

common disorder. 

Techniques: 

It is anticipated that this will involve the isolation and sorting (magnetic cell isolation 

kits and flow cytometry) immune cells from various organs from animal models of 

hypertension. In addition, assays to detect T cell derived-cytokines/chemokines 

(cytometric bead array) and reactive oxygen species generation (chemiluminescence) 

will also be utilized. 

Contacts: 

Dr Antony Vinh and Prof Rob Widdop 



Phone: 9902 4844, Rm E33 

Antony.Vinh@monash.edu 

Robert.Widdop@monash.edu 



School/Department.


MODULATION OF NEURONAL OXYTOCIN – EFFECTS ON 

SOCIAL BEHAVIOUR 

Supervisors: Dr Richard Loiacono & Dr Siew Yeen Chai 

Location: Neuropharmacology Lab & Neurophysiology Team 

Departments of Pharmacology & Physiology 


Background: 

Metallo-peptidases cleave amino acids from either the N- and C-termini of peptide 

hormones to either generate or degrade biologically active peptides. These enzymes play 

important roles in the body and alterations in their activities can impact on a diverse 

range of physiological processes in both healthy and diseased states. This project is 

focussed on one such enzyme known as insulin-regulated aminopeptidase (IRAP) or 

oxytocinase. Oxytocin is known as the social hormone, regulating complex social 

behaviours including promoting trust, pair-bonding and has been explored as potential 

therapy for social behaviour impairments observed with autism spectrum disorders. 

Aim: 

This project will investigate the behavioural phenotypes in the oxytocinase deficient 

mice to determine if regulation of endogenous oxytocin levels will affect social 

behaviour. 

Techniques: 

Behavioural testing, cell culture, immunohistochemistry 

Contacts: 

Dr Richard Loiacono & Dr Siew Yeen Chai 

Departments of Pharmacology & Physiology 


Phone: Richard 9905 4859 Siew Yeen 9905 2515 

richard.loiacono@monash.edu 

siew.chai@monash.edu 



School/Department.


ARE THE ADVERSE EFFECTS OF HORMONE REPLACEMENT 

THERAPY ON STROKE OUTCOME MEDIATED BY G PROTEIN- 

COUPLED ESTROGEN RECEPTOR SIGNALLING? 

Supervisors: Dr Brad Broughton & A/Prof Chris Sobey 

Location: Vascular Biology & Immunopharmacology Group 



Background: 

Stroke is a debilitating disease that can cause permanent neurological damage, 

complications, and death. At present, there are very few treatment options available 

for patients, thus the development of new treatments is vital to reduce the damage 

caused by stroke. In recent years, growing evidence has revealed that estrogen is 

neuroprotective against stroke, but confusingly hormone replacement therapy (HRT) 

worsens stroke outcome in post-menopausal women. Interestingly, recent work from our 

lab has found that activation of a novel estrogen receptor (G Protein-coupled Estrogen 

Receptor, GPER), which is widely distributed throughout the brain, increases cerebral 

infarct damage in males following stroke. Therefore, we hypothesise that GPER 

activity affects outcome of HRT therapy in females after stroke. 

Project aim: 

The aim of this project is to investigate the importance of GPER in mediating the 

adverse effects of HRT in stroke. The outcomes of the proposed project are 

expected to confirm that estrogen therapy has an adverse effect on stroke 

outcome in young ovariectomised mice due to activation of GPER signalling. 

Techniques: 

Techniques that will be used during this project include treating mice with estrogen and 

various GPER ligands, histochemical approaches to measure cerebral infarct damage, 

immunohistochemistry to examine the distribution of GPER in the brain and Western 

blotting to determine GPER expression levels. 

Contacts: 

Dr Brad Broughton & A/Prof Chris Sobey 



Phone: 9905 0915, Rm EG20 

bradley.broughton@monash.edu 

chris.sobey@monash.edu 

Vehicle Estradiol 



School/Department.


STEM CELL THERAPY FOR THE TREATMENT OF HIGH BLOOD 

PRESSURE 

Supervisors: A/Prof Grant Drummond & Dr Rebecca Lim 




Background: 

Hypertension (high blood pressure) is a major cause of heart failure, heart attacks and 

strokes. Over 2 million people in Australia suffer from hypertension and alarmingly, in up 

to 30% of these individuals, the condition is not controlled by current blood pressure 

medications. A recent development in the field of hypertension research is the recognition 

that the disease is caused by an influx of immune cells (i.e. T cells and macrophages) into 

the artery wall. Activation of these immune cells leads to the release of free radicals and 

cytokines that promote an inflammatory response, causing the arteries to become ‘stiff’ 

and constricted and exacerbating elevations in blood pressure. Hence, strategies that 

reduce inflammation in the vascular wall hold promise as future treatments for 

hypertension. 

Project aims: 

Stem cells have long been touted as possible cures for degenerative diseases due to their 

tissue regenerative properties. However, stem cells also display powerful antiinflammatory 

properties suggesting that they may also have therapeutic potential in 

inflammatory conditions such as hypertension. The human amniotic membrane is a readily 

available tissue (discarded after childbirth) and a rich source of stem cells. The aim of 

this project is to examine whether human amnion stem cells prevent hypertension in 

mice by exerting anti-inflammatory effects in the artery wall and in other organs 

that regulate blood pressure, such as the kidney and brain. 

Techniques: 

This project will involve surgeries on mice to induce hypertension, in vivo blood pressure 

monitoring, and the use of assays to measure immune cell numbers (flow cytometry, 

immunohistochemistry), and expression of inflammatory mediators (real-time PCR, ELISA, 

Western blotting) and free radicals (luminescence) in blood and tissues. 

Contact: 

A/Prof Grant Drummond 



Phone: 9905 4869, Rm E112 

Grant.Drummond@monash.edu 



School/Department.


EXPLORING THE VASOPROTECTIVE ACTIONS OF 

NOVEL HNO DONORS 

Supervisors: Dr Barbara Kemp-Harper 




Background: 

Nitric oxide (NO ), is an important endogenous vasodilator and regulator of vascular tone. 

In disease states such as hypertension and atherosclerosis, the NO signaling pathway is 

impaired leading to reduced blood flow to vital organs such as the brain and heart. Whilst 

nitrovasodilators can be used to overcome dysfunctional NO signaling, the clinical 

application of these agents is limited due to their susceptibility to scavenging by 

superoxide and tolerance development. 

Importantly, NO can also exist in the reduced state as nitroxyl (HNO) and recent 

evidence suggests that this nitrogen oxide is also generated endogenously and has distinct 

pharmacological properties and therapeutic advantages over NO. Thus HNO is not 

scavenged by superoxide, does not develop tolerance and can target distinct signaling 

pathways to cause vasorelaxation. As such, the bioavailability of HNO is preserved in 

disease. With a suite of vasoprotective actions, HNO donors have therapeutic potential in 

the treatment of cardiovascular disease. Currently, however the clinical utility of these 

donors is limited by their short-half lives byproducts. Excitingly, new pure HNO donors 

with longer half-lives are being developed and this study will elucidate the therapeutic 

potential of these drugs in the setting of atherosclerosis. 

Project aim: 

The aim of this honours project is to investigate the ability of novel HNO donors to serve 

as vasoprotective agents in the setting of atherosclerosis. 

Techniques: 

It is anticipated that the project will involve the use of techniques to assess the 

vasodilator (small vessel myography), anti-aggregatory and superoxide suppressing 

(chemiluminescence) actions of novel HNO donors in control and atherosclerotic mice. 

Contacts: 

Dr Barbara Kemp-Harper 



Phone: 9905 4674, Rm E140 

Barbara.Kemp@monash.edu 

Proliferation 

Platelet 

Aggregation 

HNO VSMC 

Relaxation 



School/Department. 

. - 

O 

2 

. 

O 2 

- 

. - 

O 

2 

Superoxide 

Production


EXPLORING NOVEL THERAPEUTIC STRATEGIES TO 

PREVENT ATHEROSCLEROTIC PLAQUE RUPTURE 

Supervisors: Dr Barbara Kemp-Harper & A/Prof Grant Drummond 




Background: 

Atherosclerosis is characterised by the formation of lipid filled lesions in the arterial 

wall. These plaques are relatively harmless until they become unstable and rupture leading 

to thrombus formation and vessel occlusion resulting in myocardial infarction and stroke. 

Macrophages, in particular, play a central role in plaque rupture secreting inflammatory 

molecules, reactive oxygen species (ROS) and metalloproteinases. As such, new therapies 

are sought which reduce plaque macrophage content and activation. We have evidence to 

suggest that nitroxyl (HNO), a novel redox form of nitric oxide, targets Nox2 oxidase to 

limit vascular and inflammatory cells ROS generation and may stabilize atherosclerotic 

plaques. 

Project aim: 

The aim of this honours project is to investigate the ability of HNO and NADPH oxidase 

inhibitors to serve as protective agents against atherosclerotic plaque rupture. This study 

will elucidate the potential vasoprotective effects of HNO and NAPDH oxidase inhibitors 

and may lead to the development of more effective therapies for the treatment of 

atherosclerosis. 

Techniques: 

It is anticipated that this will involve the use of assays to detect inflammation (RT-PCR, 

western blotting, immunohistochemistry), reactive oxgen species generation 

(chemiluminescence), vascular dysfunction (small vessel myography) and plaque formation 

and stability in atherosclerotic mice. 

Contacts: 

Dr Barbara Kemp-Harper and A/Prof Grant Drummond 



Phone: 9905 4674, Rm E140 

Barbara.Kemp@monash.edu 

Grant.Drummond@monash.edu 




norm 

al 

arter 

mid-stage 

atherosclero 

sis 

late-stage 

atherosclero 

sis


EXPLORING THE ROLE OF ENDOTHELIAL NF-κB ON ISCHEMIC 

STROKE OUTCOME 

Supervisors: Dr Helena Kim & A/Prof Chris Sobey 




Background: 

Stroke is the second leading cause of death worldwide and is caused by an interruption of 

cerebral blood flow by a blockage (ischemia) or a rupture (haemorrhage) in the blood 

vessel. Cerebral ischemia triggers a cascade of complex events including excitotoxicity, 

production of reactive oxygen species, up-regulation of pro-inflammatory molecules and 

transcription factors that lead to apoptosis. Inflammatory mechanisms have come into the 

focus of stroke research because they contribute substantially to secondary brain damage. 

Nuclear factor kappa B (NF-κB) is a transcription factor that modulates gene expression 

of a range of cytokines and other pro-inflammatory molecules. Recent findings suggest 

that inhibition of NF-κB, especially those expressed in neurons, provide neuroprotection 

following stroke. 

Project aim: 

The aim of this honours project is to investigate the role of endothelial NF-κB on stroke 

outcome using mice overexpressing NF-κB inhibitor, IκBα. This study will elucidate the 

potential effect of NF-κB inhibition and blocking of further inflammatory response 

following stroke and may lead to the development of more effective therapies for the 

treatment of stroke. 

Techniques: 

This project will involve the use of assays to measure behavioural deficits (hanging wire 

test, neurological score), brain infarct size (thionin stain, Image J software) and immune 

responses (immunohistochemistry) in IκBα-overexpressing mice following stroke. 

Contacts: 

Dr Helena Kim and A/Prof Chris Sobey 



Phone: 9905 1146, Rm E139 

Helena.Kim@monash.edu 

Chris.Sobey@monash.edu 



School/Department.


INVESTIGATING MECHANISMS LINKING HYPERTENSION AND 

COGNITIVE IMPAIRMENT 

Supervisors: Dr Alyson Miller 




Background: 

As the population of the world ages, the incidence of cognitive impairment and dementias 

is expected to rise exponentially. In fact, it is predicted that the number of dementia 

patients worldwide will quadruple over the next three decades to reach approximately 81 

million by 2040. Although Alzheimer disease (AD) is the most commonly diagnosed cause 

of cognitive dysfunction among the aged, cognitive impairment caused by cardiovascular 

diseases are also important independent causes and contributors to cognitive dysfunction. 

For example, there is growing evidence hypertension during mid-life increases the risk for 

cognitive impairment later in life. Despite this evidence, the mechanistic interactions 

between hypertension and cognitive dysfunction are still an enigma. 

Project aims: 

The aims of this Honours project are to firstly verify that hypertension causes cognitive 

dysfunction. Secondly, this project will explore potential pathways and mediators that 

mechanistically link hypertension and cognitive impairment, with a particular focus on 

oxidative and inflammatory processes. 

Techniques: 

Techniques that will be used during this project include behavioural tests to assess 

cognitive function, small vessel myography to assess vascular function, chemiluminescence 

based assays to assess oxidative stress, and molecular approaches and FACS analysis to 

assess inflammation. 

Contact: 

Dr Alyson Miller 


Phone: 9905 3817 

Alyson.Miller@monash.edu 



School/Department.


NOVEL PHARMACOLOGICAL TARGETS FOR INFLUENZA A VIRUS 

INDUCED LUNG INFLAMMATION 

Supervisors: Dr Stavros Selemidis 

Dr Ross Vlahos 

Location: Department of Pharmacology, Monash University, Clayton and 

Department of Pharmacology, Melbourne University. 

Background: 

Current drug therapies to treat influenza pathology are focused primarily on halting 

mechanisms of viral infection and replication. Far less attention has been directed to 

identifying immunopathological mechanisms of host defence that lead to the acute lung 

injury. Animal and human studies provide compelling evidence that the acute lung injury 

following influenza A virus infection is caused by reactive oxygen species (ROS) 

production such as superoxide anion. Importantly, the primary sites of influenza A virus 

infection, namely airway epithelial cells, and resident and infiltrating macrophages are key 

sites of ROS production via Nox1- and Nox2-containing NADPH oxidases, respectively. 

Our preliminary studies have identified the Nox1 and Nox2 enzymes as novel targets for 

modulating pathology irrespective of the infecting influenza strain. 

Project aims: The aim of this honours project is to investigate the roles of NADPH 

oxidases in influenza A virus induced lung injury and inflammation. To achieve this a 

multidisciplinary honours project has been devised. It is anticipated that these studies 

will have a major impact on modern drug discovery focusing on oxidative stress caused by 

influenza A viruses. 

Techniques: 

It is anticipated that this will involve the use of cell culture and in vivo animal models; and 

assays to detect NADPH oxidase expression and localization (western blotting, 

immunohistochemistry), reactive oxygen species generation (chemiluminescence) and 

inflammatory cell characterization (flow cytometry). 

Contacts: 

Dr Stavros Selemidis 



Phone: 9905 5756, Rm E137 

Stavros.selemidis@monash.edu 



School/Department.


UNDERSTANDING THE BIOLOGY OF REACTIVE OXYGEN 

SPECIES IN CANCER 

Supervisors: Dr Stavros Selemidis 

Location: Department of Pharmacology 


Background: 

The socio-economic burden of prostate cancer is unquestionable claiming many lives 

worldwide. Angiogenesis is necessary for tumours to grow and metastasise, and much 

effort has been directed towards identification of the genetic and molecular mechanisms 

that lead to angiogenesis and in the development of pharmacological agents to halt 

angiogenesis. It has long been known that tumour and endothelial cell proliferation and 

survival are crucial for a sustained angiogenic process. Emerging evidence implicates 

reactive oxygen species (ROS) such as superoxide and hydrogen peroxide (H2O2) to play a 

fundamental role in facilitating angiogenesis. The primary sources of ROS are the Nox2- 

and Nox4-containing NADPH oxidases. 

Project aims: The aim of this honours project is to investigate the roles of NADPH 

oxidases in prostate tumour-associated angiogenesis. The findings from this study may 

provide a new understanding of the molecular mechanisms that underpin angiogenesis, and 

a rationale for the combined suppression of NADPH oxidases to inhibit angiogenesis in 

prostate cancer. 

Techniques: 

It is anticipated that this will involve the use of cell culture and in vivo animal models; and 

assays to detect NADPH oxidase expression and localization (western blotting, 

immunohistochemistry), reactive oxygen species generation (chemiluminescence) and 

immune cell characterization (flow cytometry). 

Contacts: 

Dr Stavros Selemidis 



Phone: 9905 5756, Rm E137 

Stavros.selemidis@monash.edu 

Figure: Preliminary data from our laboratory 

demonstrating a significant reduction in 

prostate tumour development in a NADPH 

oxidase-deficient mouse (i.e Nox2 isoform) 

compared to a WT control mouse. 



School/Department.


VENOMS AND TOXINS PROJECTS 

Supervisors: Professor Wayne Hodgson 

Location: Monash Venom Group 



Background: 

The Monash Venom Group investigates the toxinology of venomous snakes, spiders and 

sea creatures. 

Projects: 

1. Molecular toxinology of Australia’s lesser known venomous snakes 

2. Profiling Australian and Malaysian snake venoms to guide treatment strategies 

Techniques: 

Venom fractionation, HPLC, organ bath experimentation, blood coagulation assays, 

myotoxicity assays 

Contacts: 

Professor Wayne Hodgson 



Phone: 9905 4861 

Wayne.Hodgson@monash.edu 



School/Department.


DEFINING THE FUNCTION OF THE HUMAN PLATELET 

THROMBIN RECEPTOR, PAR4, IN ARTERIAL THROMBOSIS 

Supervisors: Dr Justin Hamilton & A/Prof Grant Drummond 

Location: Australian Centre for Blood Diseases 

Alfred Medical Research & Education Precinct 

Alfred Hospital, Prahran 

Background: 

Arterial thrombosis causes heart attack and stroke is the leading cause of death and 

disability in most countries. Platelets are the blood cells responsible for forming arterial 

thrombi, and new drugs which block this function of platelets are sought for effective 

antithrombotic therapy. We have shown that thrombin activates platelets via proteaseactivated 

receptors (PARs) and that PAR-deficient mice are protected against thrombosis 

but do not exhibit spontaneous bleeding (see Figure). Our work has led to the 

development of PAR antagonists – two of which are currently in Phase 3 clinical trials for 

the prevention of arterial thrombosis. However, there are two thrombin receptors on 

human platelets, PAR1 and PAR4, and current antagonists target only PAR1. Indeed, little 

is known about PAR4. To address this, we have recently developed the first specific PAR4 

antagonist which will allow us for the first time to determine the role of PAR4 in human 

platelet function. 

Project aims: 

This project will examine the contribution of PAR4 to human platelet activation and 

thrombus formation and will help determine whether PAR4 is a suitable candidate for the 

development of anti-thrombotic therapies. 

Techniques: 

The project will utilise many useful techniques including functional experiments on isolated 

platelets, ex vivo whole blood thrombosis experiments, confocal microscopy, and in vivo 

mouse models of thrombosis and haemostasis, and will appeal to students interested in 

researching and developing future therapies for heart attack and stroke. 

Contact: 

Dr Justin Hamilton 

Australian Centre for Blood 

Diseases 


Phone: 9903 0125 

Justin.Hamilton@monash.edu 

In vivo thrombosis: Shown are platelets (red) in 

arterioles of control or PAR4-deficient mice at various 

times after laser-induced vascular injury. Note the 

limited growth of the thrombus in PAR4-deficient mice. 



School/Department.


WHAT TURNS YOU ON? USING A NOVEL RNAi-BASED 

APPROACH TO DETERMINE PLATELET ACTIVATION SIGNALS 

IN THROMBOSIS 

Supervisors: Dr Justin Hamilton & A/Prof Grant Drummond 

Location: Australian Centre for Blood Diseases 

Alfred Medical Research & Education Precinct 

Alfred Hospital, Prahran 

Background: 

Thrombosis is the formation of blood clots leading to heart attack and stroke. Platelets 

are the blood cells which form these clots, but they must be activated before they will do 

so. Therefore we are interested in determining what controls the activation of platelets 

during blood clot formation. We have recently established a novel transgenic strategy for 

inducible knockdown of genes of interest in mouse platelets in vivo. We are now using this 

approach to perform the first studies on the intracellular signalling enzyme, PI3K-C2α, in 

platelet function. We have shown that PI3K-C2α-deficient mice have dysregulated 

platelet function in vitro and in vivo (e.g. see figure). 

Project aims: 

This project will use our novel RNAi-based mouse genetic model to further define the 

contribution of PI3K-C2α to platelet function to determine whether inhibition of PI3K- 

C2α is an effective strategy for the prevention of arterial thrombosis. 

Techniques: 

The project will use a range of techniques, including a novel mouse genetic approach, 

molecular biology (RT-PCR, Western blotting, etc), as well as basic techniques in blood 

handling and platelet preparation and manipulation and will suit students interested in 

gaining experience in a variety of laboratory techniques and those interested in 

researching a potential new drug target for the prevention of heart attack and stroke. 

Contact: 

Dr Justin Hamilton 

Australian Centre for Blood Diseases 


Phone: 9903 0125 

Justin.Hamilton@monash.edu 

PI3K-C2α-deficient mice do not clot as 

efficiently as control mice. The time taken 

for mice to stop bleeding following a defined 

wound cut into their tail was recorded. 

Wild-type PI3K-C2α 

mutant 



School/Department.


THE ROLE OF AT2R IN MACROPHAGE POLARIZATION 

Supervisors: Prof Jaye Chin-Dusting, Dr Jennifer Irvine & 

Prof Rob Widdop 

Location: Vascular Pharmacology 


Prahran 

Background: 

Atherosclerosis has been identified as an underlying cause of cardiovascular disease 

(CVD). Vascular inflammation, a critical early event in the development of atherosclerosis, 

involves the recruitment and adhesion of monocytes to the endothelium. The monocytes 

then transmigrate through the vessel wall into tissue, where they then differentiate into 

macrophages. Under physiological conditions, the monocyte-derived macrophage is of the 

M2 type playing a normal protective role in the organism. When pathological factors 

(including inflammation) are present, the differentiated macrophage possesses features 

which damage tissue, and this type of macrophage is normally termed M1. M1 macrophages 

play an important role in CVD. 

The Renin-Angiotensin System (RAS) regulates cardiovascular homeostasis largely via 

activation of the angiotensin type 1 receptor (AT1R), which exhibits, amongst others, proinflammatory 

effects. However, despite widespread use of AT1R blockers in the 

treatment of CVD diseases such as hypertension, many patients still exhibit endothelial 

dysfunction and remain at risk of death from complications such as heart attack and 

stroke. Less well established, the angiotensin type 2 receptor (AT2R) and the Mas 

receptor (MasR) subtypes are thought to counter-regulate the effects of the AT1R. 

Project aims: 

This project aims to explore the potential effects of direct AT2R activation on 

macrophage polarization. We will isolate monocytes from human blood, differentiate them 

into macrophages, and induce the macrophages into M1 or M2 states respectively. We will 

then study the influence of AT2R activation on the induction of M1 or M2 macrophages 

and AT2R-induced alteration of macrophage polarization. 

Techniques: 

This project involves a range of technologies of cellular and molecular biology, such as cell 

culture, flow cytometry, real-time PCR, western blot and cellular imaging etc. 

Contacts: 

Prof Jaye Chin-Dusting & Dr Jennifer Irvine 

Vascular Pharmacology 


jaye.chin-dusting@bakeridi.edu.au, jennifer.irvine@bakeridi.edu.au 



School/Department.


EFFECTS OF POSITIVE ALLOSTERIC MODULATOR OF GABAA 

RECEPTORS ON HYPERTENSION AND STRESS 

Supervisors: Professor Geoffrey A. Head, Dr Pamela Davern 

Location: Neuropharmacology Laboratory 

BakerIDI Heart & Diabetes Institute 

Prahran 

Background: 

Allopregnanolone (AlloP), a neurosteroid produced in the brain is a potent endogenous 

positive allosteric modulator of GABA action at GABAA receptors. Increased NPY Y1 

receptor gene expression in the amygdala is a major mechanism of AlloP anxiolytic 

effects. Systemic administration of AlloP or drugs that reverse the reduction of AlloP (eg. 

Fluoxetine) in the brain induced by social isolation, reduce anxiety and aggressive 

behavioural abnormalities. However, the cardiovascular consequences of the direct actions 

on stress have been largely overlooked. We suggest that Schlager hypertensive mice may 

be deficient in production of AlloP leading to the dysfunction of the GABAA receptor and 

that this can be reversed by AlloP administration. This will determine whether the defect 

is reversible or not. The latter would suggest the genetic differences are a primary 

deficit rather than induced by a lifetime of heightened stress 

Project aims: 

The aim is to determine the effect of chronic systemic administration of AlloP on blood 

pressure and response to stress in BPH and BPN mice. 

Techniques: 

The project involves radiotelemetry measurement of blood pressure in conscious mice, 

behavioural testing using a number of mild stressors such as restraint and 

immunohistochemistry to examine thec ahnges in activation of neuropeptide Y and GABA 

neurons in the amygdala and hypothalamus. 

Contacts: 

Prof. Geoffrey Head and Dr Pamela Davern 



Phone: 03 8532 1332 

Geoff.head@bakeridi.edu.au 

Pamela.Davern@bakeridi.edu.au 

In BPH hypertensive mice many neurons expressing GABAA 

receptors (orange) were also immunoreactive for NPY (red) and 

each of these were triple labeled with Fos (yellow) in the 

paraventricular nucleus (PVN) and medial amygdala (MeAm). 



School/Department.


ROLE OF BRAIN PATHWAYS IN CHRONIC STRESS 

Supervisors: Professor Geoffrey A. Head, Dr Pamela Davern 

& Sandra Burke 



Prahran 

Background: 

There is now strong evidence to suggest that the sympathetic nervous system (SNS) 

makes a significant contribution to certain forms of hypertension. One of the suggested 

mechanisms through which the SNS may be “activated” in hypertension is through the 

renin‐angiotensin system. We have recently demonstrated that a very low “subpressor” 

dose of Angiotensin given for several months to rabbits can produce a very modest 

increase in blood pressure but appears to amplify the effects of chronic stress. Using 

immuno‐histochemistry, we have found that specific areas of the brain are activated by 

infusing angiotensin, an action which is mediated through areas of the brain that does not 

have a blood brain barrier. These central pathways appear to be “sensitised”, reflecting a 

type of positive feed forward CNS plasticity. 

Project aims: 

We now wish to determine which areas of the brain and in particular the hypothalamus 

may be responsible for this effect on chronic stress 

Techniques: 

The project will involve some animal surgery, experiments to measure blood pressure 

before and during a relatively mild air jet stress and then perfusion fixation of the brain 

to process for immunohistochemistry. The use of specific antibodies, fluorescence and 

confocal imaging will then be used to identify brain regions and specific neurochemical 

involved in the response to acute and chronic stress. 

Contacts: 

Prof. Geoffrey Head and Dr Pamela Davern 



Phone: 03 8532 1332 



Fos Immunohistochemistry: Effect of 

Chronic Stress and 6wks low dose Ang II 



School/Department.


WHICH PARTS OF THE BRAIN ARE AFFECTED IN OBESITY 

RELATED HYPERTENSION? 

Supervisors: Professor Geoffrey A. Head, Dr Pamela Davern and 

Dr Joon Lim 



Prahran 

Background: 

Cardiovascular disease and its ultimate consequences are a significant burden on health 

systems in Australia and around the developed world. It is increasingly apparent that the 

high rate of hypertension in many societies can be attributed to an equally alarming rate 

of obesity. Despite this link, there is a less than complete understanding of the manner by 

which obesity alters central nervous system function to result in hypertension. It is known 

that the sympathetic nervous system (SNS), responsible in part for the control of blood 

pressure and is overactive in obese humans. It is also known that several peripheral 

signalling chemicals act at specific brain sites to modulate both appetite and also the 

activity of the SNS . For example, leptin, produced by fat and responsible for reducing 

appetite by acting at specific sites in the brain may act also to either stimulate or inhibit 

SNS activity depending on which neurons it acts at in the hypothalamus. Nonetheless, 

despite a comprehensive understanding of changes in appetite systems in the brain in 

obesity, and a known link between some of these appetite centres and other nuclei that 

control SNS outflow there is little understanding of how the sympathetic nervous system 

is activated during the development of obesity. 

Project aims: 

We seek to determine the manner by which various brain centres respond in obesity by 

studying patterns of neural activation and measuring SNS activity and blood pressure 

during the development of obesity using rabbits. 

Techniques: 

This project will involve surgical implantation of telemetric probes (for measurement of 

renal sympathetic nerve activity and haemodynamic variables) and guide cannulae into 

distinct brain regions to allow injection of drugs. Students will also learn 

immunohistochemical techniques in order to identify by immunofluorescent labelling those 

neurons that are responsible for the genesis of obesity related hypertension. 

Contacts: 

Prof. Geoffrey Head, Dr Pamela 

Davern & Dr Joon Lim 


Phone: 03 8532 1332 


Joon.Lim@bakeridi.edu.au 


NITROXYL, A RELATIVE OF 



School/Department.


HNO, IS A NATURALLY-OCCURRING CARDIOPROTECTIVE 

MOLECULE 

Supervisors: A/Prof Rebecca Ritchie & Dr Barbara Kemp-Harper 

Location: Heart Failure Pharmacology Laboratory 


75 Commercial Rd, Melbourne 

Background: 

The nitric oxide (NO•)/cGMP signalling system is as a powerful cardiac antihypertrophic 

mechanism. Nitroxyl (HNO), a novel redox sibling of NO•, has several therapeutic 

advantages for the treatment of cardiovascular diseases. The nitric oxide (NO•)/cGMP 

signalling system is as a powerful cardiac antihypertrophic mechanism. Nitroxyl (HNO), a 

novel redox sibling of NO•, has several therapeutic advantages for the treatment of 

cardiovascular diseases. We have shown that HNO prevents cardiomyocyte hypertrophy 

(abnormal pathological growth) and generation of superoxide, while concomitantly 

enhancing cardiac function (the latter in direct contrast to NO•). 

Project aims: 

This project explores whether HNO pharmacotherapy limits myocardial dysfunction, 

induced by hypertension, heart failure or diabetes, and the mechanisms for these actions. 

Putative independent mediators of chronic HNO cardioprotection include cGMP-mediated 

suppression of reactive oxygen species (ROS), and thiol-mediated preservation of cardiac 

calcium handling protein activity (e.g. SERCA2a, RyR2), whose activity is abnormally 

affected in cardiac pathologies. This project will be tailored depending on the student’s 

abilities and interests. The outcome of this project will be definitive information 

regarding the mechanism(s) and effectiveness of HNO-mediated rescue of myocardial 

dysfunction. Ultimately, HNO-based strategies may offer new treatment options for 

cardiac disease, either alone or on top of standard care. 

Techniques: 

This project will provide the opportunity for learning a range of techniques, including cell 

culture (cardiomyocytes and/or cardiac fibroblasts), pharmacological (e.g. isolated rodent 

hearts ex vivo or in vivo) models of cardiac disease for assessing cardiac function and 

blood pressure, biochemical (Westerns, ROS detection, ELISA, real-time PCR) and/or 

histological techniques. 

Contact: 

A/Prof Rebecca Ritchie 

Heart Failure Pharmacology Laboratory 


Phone: 8532 1392 

Email: rebecca.ritchie@bakeridi.edu.au 

Heart disease (e.g. 

Hypertension, Diabetes) 

↑LV hypertrophy 

↑LV fibrosis 

X X 

↓ LV 

function 

↓SERCA/RyR 

function 

X 

↑SERCA/RyR 

function 

↑ cGMP thiol reactivity 




↑ROS 

Nitroxyl 

heart failure 

death 

X ?


NEW STRATEGIES TO RESCUE 

DIABETES-INDUCED CARDIAC DYSFUNCTION 

Supervisors: A/Prof Rebecca Ritchie 




Background: 

Diabetes is Australia’s fastest growing chronic disease; one million Australians have been 

diagnosed, with close to one million more yet to be identified. Diabetes impairs left 

ventricular (LV) function, increasing the risk of death from heart failure > 2-fold. New 

therapies for restoring cardiac function in the diabetic heart are thus highly desirable. 

The aetiology of diabetic heart disease is distinct from other causes of LV dysfunction, as 

it is characterised initially by diastolic dysfunction, where relaxation of the cardiac 

muscle following contraction is prolonged. Our laboratory has demonstrated that 

antioxidant and/or ROS-suppressing approaches, as well as activation of cardioprotective 

signalling and negative regulators of LV hypertrophy, are beneficial for treating the 

cardiac complications of type 1 and type 2 diabetes in the intact heart. 

Project aims: 

This project explores a novel potential therapeutic strategy for rescuing cardiac function 

and structure in the diabetic heart, determining whether post-translational protein 

modifications induced by high glucose play a causal role in development of diabetic 

cardiomyopathy, and investigate whether pharmacological and/or gene-based strategies 

targeted at limiting these modifications can prevent diabetes-induced LV dysfunction, LV 

fibrosis, hypertrophy and excess generation of reactive oxygen species (ROS) such 

superoxide. Ultimately, treatment strategies that may emerge from these studies may 

provide significant benefits alone or in combination with current standard care, to 

ultimately reduce progression to heart failure and death in diabetic patients. 

Techniques: 

This project will provide the opportunity for learning pharmacological (e.g. isolated rodent 

hearts ex vivo or in vivo) models of cardiac disease for assessing cardiac function and 

blood pressure, biochemical (Westerns, ROS detection, ELISA, real-time PCR) and/or 

histological techniques. 

Contact: 




Phone: 8532 1392 




School/Department.


TARGETING ANTI-INFLAMMATORY PROTEIN ANNEXIN-A1 

FOR PROTECTION FROM MYOCARDIAL INFARCTION 

(HEART ATTACK) 

Supervisors: A/Prof Rebecca Ritchie 




Background: 

Myocardial infarction (a sustained impairment in coronary bloodflow) and the resultant 

heart failure is a major cause of death in Western societies. The therapeutic potential of 

the endogenous anti-inflammatory mediator annexin-A1 (ANX-A1) has been recognized in a 

range of inflammatory disorders. We have shown that ANX-A1 has powerful protective 

actions against cardiac injury and loss of LV contractile function. This project explores 

the potential for ANX-A mimetics to reduce cardiac ischaemia-reperfusion injury. 

Project aims: 

The project will test the hypothesis that ANX-A1 represents a novel modulator of 

myocardial viability and LV contractile function following ischaemia-reperfusion, and will 

seek to investigate the cardioprotective function of endogenous ANX-A1 in I-R injury, the 

receptors responsible for cardioprotection elicited by ANX-A1 and its mimetics (likely the 

formyl peptide receptor family, FPR1 and/or FPR2), and examine the potential therapeutic 

opportunities offered by exogenous ANX-A1 mimetics after I-R injury in the intact heart. 

Development of therapeutic strategies for treating myocardial infarction, alone or 

concurrent with standard care, will ultimately reduce progression to heart failure and 

death in affected patients. 

Techniques: 

The project provides the opportunity for learning a range of techniques, including cell 

culture, as well as physiological ex vivo and/or in vivo models of cardiac ischaemia for 

studying cardiac function and structure, biochemical (and/or histological techniques. 

Contact: 




Phone: 8532 1392 


↓ inflammation 

in vivo 

preservation of 

cardiac morphology 

ANX-A1 mimetics 

FPR2 FPR1 

G q/11 G 

↑ myocardial 

viability 

heart failure 

death 

early rescue of 

contractile function 

preservation of 

contractile function 




X 

P-Akt 

G i/o 

X


SIGNALLING BIAS OF CALCITONIN RECEPTOR POLYMORPHS 

Supervisors: Dr Sebastian Furness and Prof Patrick Sexton 

Location: Drug Discovery Biology theme 

Monash Institute of Pharmaceutical Sciences 

Monash University, Parkville 

Background: 

The Calcitonin receptor (CTR) is involved in calcium homeostasis by regulating its secretion 

in kidneys and the resorbtion of bone by osteoclasts in response to Calcitonin. The CTR is 

polymorphic in the C-terminal tail with a number of epidemiological studies correlating this 

polymorphism with osteoporotic risk. One previous study failed to uncover differences 

between CTR polymorhs. We have used several model cell backgrounds to analyse 

differences in signaling arising from these polymorphs. We have shown that there are a 

number of polymorphism-dependant changes in signaling efficacy toward adenylate 

cyclase, extracellular signal-regulated kinase (ERK) phosphorylation and intracellular 

calcium release. Further, we have demonstrated that these signaling biases are dependent 

on cellular background. 

Project aim: 

This project will extend the existing signalling bias studies using bioluminescence 

resonance energy transfer (BRET) to examine the interaction between CTR and members 

of the arrestin and GRK families. 

Techniques: 

Molecular Biology: basic molecular biology procedures including isolation and preparation of 

DNA, cell culture and cell transfection. Protein-protein interaction will be measured by 

resonance energy transfer and via fluorescence imaging. Pharmacological characterization 

of receptors will be performed using high-throughput assays for measurement of 

intracellular signaling including cAMP formation, ERK phosphylation and intracellular 

calcium mobilization. 

Contacts: 

Dr Sebastian Furness 

Phone: 9903 9055, Rm 5-09, Building 404; 

Sebastian.furness@monash.edu 



School/Department.


COMPARTMENTALISATION OF RELAXIN-STIMULATED 

SIGNALLING IN SINGLE CELLS 

Supervisors: Dr Michelle Halls & Prof Roger Summers 




Background: 

Relaxin is a pleiotropic hormone that is implicated in multiple physiological systems, and 

has therapeutic potential in heart failure (currently in phase III clinical trials), fibrosis 

and cancer metastases. Relaxin binding to its G protein - coupled receptor, RXFP1, induces 

the activation of multiple signalling pathways. The major consequence of receptor 

stimulation is increased cAMP production, which occurs via a complex, simultaneous 

activation of three distinct G protein pathways: Gαs stimulation of adenylyl cyclase (AC) 

activity to increase cAMP, GαoB inhibition of AC, and Gαi3 -activation of AC. 

The activation of three separate pathways to independently affect the same second 

messenger, suggests that relaxin-stimulated cAMP production is compartmentalised within 

the cell. Compartmentalisation, or the specific localisation of signalling mediators into 

discrete regions of the cell, allows cells to specifically tailor a physiological response to a 

distinct stimulus. 

Project aim: 

This project will explore the extent of signalling compartmentalisation following relaxin 

activation of RXFP1. Specifically, we will use state-of-the-art biosensors, which can be 

targeted to different areas of the cell, to examine the compartmentalisation of cAMP, 

protein kinase C (PKC) and extracellular signal regulated kinase ( -RK) phosphorylation in 

single live cells. Information gained from this project will demonstrate the importance of 

signalling compartmentalisation in the generation of a defined physiological response. 

Further, understanding the compartmentalisation of relaxin -mediated signalling may make 

clear some of the clinically relevant effects of relaxin in conditions such as heart failure. 

Techniques: 

Molecular'Biology: basic molecular biology procedures including isolation and preparation 

of DNA, cell culture and cell transfection. Single-Cell Imaging: cutting edge, high 

throughput imaging will be used to measure activation of cAMP and phosphorylation of PKC 

and ERK by the use of fluorescence -resonance energy transfer (FRET) -based biosensors 

targeted to the plasma membrane, bulk cytosol or nucleus of single cells. Pharmacological: 

the intracellular signalling pathways leading to each of the compartmentalised responses 

will be elucidated by the use of pharmacological inhibitors of signalling. 

Contacts: 

Dr Michelle Halls 

Phone: 9903 9094, Rm 324, Building 404 

michelle.halls@monash.edu 



School/Department.


MOLECULAR MECHANISMS OF A NOVEL ALLOSTERIC 

MODULATOR OF THE D2-LIKE DOPAMINE RECEPTORS 

Supervisors: Dr Rob Lane, Prof Arthur Christopoulos 




Background & Project Details: 

The neurotransmitter dopamine mediates its action via five G protein coupled receptors 

(the dopamine receptors D1-5R) and been shown to play a vital role in a range of central 

nervous system functions including voluntary movement, reward, memory and learning. 

Dysregulation of dopamine signalling is associated with many human disorders, including 

schizophrenia and Parkinson’s disease. 

Although, antipsychotic drug discovery at the dopamine receptors has focused on 

targeting the binding site for endogenous dopamine (the ‘orthosteric’ site) this approach is 

associated with severe extrapyramidal side-effects and poor receptor subtype selectivity. 

Targeting a spatially distinct ‘allosteric’ site on the receptor may offer greater subtype 

selectivity with fewer side-effects. Recently, the first drug-like allosteric ligand of the 

dopamine receptor was described. We have demonstrated that this ligand adopts a 

simultaneous orthosteric/allosteric (bitopic) mode of binding at the dopamine D2 

receptor. However, to enable us to develop novel improved modulators of this receptor we 

need to understand how this drug exerts its modulatory effect. 

To determine the mechanism and site of action of this novel drug, we will use a combined 

approach. First, we have generated a library of receptors in which we have mutated amino 

acids within the receptor that have been implicated in the binding of both orthosteric 

ligands or potential allosteric ligands. Second, we have synthesised a number of 

derivatives of this drug in which key parts of the drug have been modified. These mutant 

receptors and novel drugs will be characterized using a number of state of the art 

functional assays. Information gained from this project and the recently published crystal 

structure of the dopamine D3 receptor can be used to predict the mode of binding of this 

receptor and will be the starting point towards the development of novel allosteric drugs 

for this important therapeutic target. 

Techniques: 

Structural: Site-directed mutagenesis and molecular modeling, to determine the binding 

mode of SB269652. Pharmacological: Receptor function will be quantified by downstream 

signalling using state-of-the-art, high throughput assays of extracellular signal regulated 

kinases (ERK1/2), intracellular cAMP, and resonance energy transfer techniques to detect 

interaction with β arrestin. Molecular biological: As part of the project you will also learn 

basic molecular biological procedures including isolation and preparation of DNA, cell 

culture and cell transfection. 

Contacts: 

Dr Rob Lane 


rob.lane@monash.edu 



School/Department.


PROBING AN ALLOSTERIC BINDING SITE IN THE HUMAN 

CASR 

Supervisors: Dr Katie Leach, Dr Laura Lopez, Prof Arthur Christopoulos 





The human calcium sensing receptor (CaSR) is a family C G protein coupled receptor 

(GPCR) that plays a pivotal role in extracellular calcium (Ca2+o) homeostasis and 

parathyroid hormone (PTH) secretion. It is expressed on the surface of a variety of cell 

types throughout the body, where it responds to small increases in free Ca2+o 

concentrations. Over 200 clinically relevant polymorphisms have been identified in the 

CaSR and unfortunately they can result in a number of disorders, such as autosomal 

dominant hypocalcaemia (ADH), Bartter syndrome type V, familial hypocalciuric 

hypercalcaemia (FHH), familial benign hypercalcaemia (FBH) and neonatal severe 

hyperparathyroidism (NSHPT). These disorders are generally characterised by an 

imbalance of PTH and/or Ca2+o. 

Cinacalcet, a positive allosteric CaSR modulator prescribed for the treatment of primary 

and secondary hyperparathyroidism was recently shown to successfully normalise serum 

Ca2+o in individuals with loss-of-function CaSR mutations, suggesting that cinacalcet and 

other allosteric CaSR modulators could be invaluable in the treatment of disorders linked 

to CaSR mutations. Recent molecular modelling and mutagenesis studies have predicted 

that the binding site(s) for allosteric CaSR modulators comprises residues located in the 

transmembrane (TM) domains and extracellular loops of the receptor, where a growing 

number of naturally occurring mutations are being identified. However, although 

mutagenesis studies, which have involved alanine substitution of amino acids predicted to 

interact with allosteric drugs, have shown effects on the actions of allosteric modulators 

at the CaSR, no study has differentiated between effects of these mutations on the 

binding affinity of the allosteric modulator, versus the cooperativity of the modulator; 

that is to say the ability of the drug to modulate the actions of the endogenous agonist, 

Ca2+o. For instance, mutation of Iso841 to Ala (I841A) in the 7th TM domain of the 

receptor was previously reported to attenuate the binding affinity of the negative 

allosteric modulator, Calhex 231, due to effects of the mutation on the ability of Calhex 

231 to reduce Ca2+o signalling. However, we have shown that these mutational effects are 

driven by changes in the cooperativity between Ca2+o and the negative allosteric 

modulator, whilst the binding affinity of the drug is unaltered at the I841A mutant. Thus, 

it is imperative that mutagenesis data be interpreted correctly if we want to define the 

binding site of allosteric CaSR drugs. 

Techniques: Cell culture: This project requires routine growth and culturing of human 

kidney embryonic (HEK293) cell lines expressing the wild type and headless human CaSRs. 

Pharmacological: Receptor function will be quantified by downstream signalling using high 

throughput assays that measure extracellular signal regulated kinases (ERK1/2), 

intracellular Ca2+o mobilisation and inositol phosphate accumulation. Structural: Alanine 



School/Department.


substitution of amino acids that have either previously been implicated in the binding of 

allosteric CaSR modulators, or that have been identified from a recent investigation into 

the effects of naturally occurring amino acids performed in our laboratory, will be used to 

determine the involvement of amino acids in the function of allosteric modulators. This 

information will be used to aid the design of more accurate molecular models of the CaSR, 

which are currently lacking. 

Contacts: 

Dr Katie Leach 


Katie.leach@monash.edu 



School/Department.


ALLOSTERIC PHARMACOREGULATION OF A HEADLESS CASR 

Supervisors: Dr Katie Leach, Prof Arthur Christopoulos 





The human calcium sensing receptor (CaSR) is a family C G protein coupled receptor 

(GPCR) that plays a pivotal role in extracellular calcium (Ca2+o) homeostasis and 

parathyroid hormone (PTH) secretion. It is expressed on the surface of a variety of cell 

types throughout the body, where it responds to small increases in free Ca2+o 

concentrations. Although the primary Ca2+- (orthosteric) binding site has been localised 

to the large extracellular N-terminal region of the receptor [1-3], otherwise known as the 

venus flytrap (VFT) domain, evidence has pointed towards an additional binding site within 

the transmembrane spanning regions of the receptor [4-5]. This second site is 

topographically distinct from the region that binds allosteric calcilytics and calcimimetics 

[5], which modulate the activity of Ca2+ o at the CaSR, although it is unclear which amino 

acids form this second binding site for Ca2+ o. Thus, the aim of this study is to 

investigate the activity of orthosteric and allosteric ligands at a “headless” CaSR that 

lacks the first 599 amino acids that comprise the VFT domain and is thus composed of the 

TM domains and a functional but truncated C-terminal tail, which lacks the last 176 amino 

acids of the receptor. The extent to which allosteric modulators retain their ability to 

regulate CaSR activity in the absence of the primary Ca2+o-binding site will be 

investigated to gain an understanding of the mechanism of allosteric modulation at the 

human CaSR. 

1. Huang, Y., Zhou, Y., Castiblanco, A., Yang, W., Brown, E. M., and Yang, J. J. (2009) 

Biochemistry 48, 388-398 

2. Huang, Y., Zhou, Y., Yang, W., Butters, R., Lee, H. W., Li, S., Castiblanco, A., Brown, 

E. M., and Yang, J. J. (2007) J Biol Chem 282, 19000-19010 

3. Silve, C., Petrel, C., Leroy, C., Bruel, H., Mallet, E., Rognan, D., and Ruat, M. (2005) 

J Biol Chem 280, 37917-37923 

4. Ray, K., Tisdale, J., Dodd, R. H., Dauban, P., Ruat, M., and Northup, J. K. (2005) J 

Biol Chem 280, 37013-37020 

5. Ray, K., and Northup, J. (2002) J Biol Chem 277, 18908-18913 

Techniques: Cell culture: This project requires routine growth and culturing of human 

kidney embryonic (HEK293) cell lines expressing the wild type and headless human CaSRs. 

Pharmacological: Receptor function will be quantified by downstream signalling using high 

throughput assays that measure extracellular signal regulated kinases (ERK1/2), 

intracellular Ca2+o mobilisation and inositol phosphate accumulation. 

Contacts: 

Dr Katie Leach 


Katie.leach@monash.edu 



School/Department.


INVESTIGATION OF MUSCARINIC M4 RECEPTOR FUNCTION 

USING LABEL FREE IMPEDANCE TECHNOLOGY 

Supervisors: Dr Patricia Rueda, Prof Arthur Christopoulos, Dr Chris Langmead 





The muscarinic M4 receptor plays a role in a range of central nervous system functions 

including voluntary movement, reward, memory and learning. The metabotropic actions of 

acetylcholine are mediated by its interaction with five G protein-coupled receptors 

(GPCRs), the muscarinic M1-M5 receptors. Because acetylcholine is involved in such critical 

physiological processes, it is not surprising that many human disorders, including 

schizophrenia, Parkinson’s disease and Alzheimer’s disease, have been related to 

cholinergic dysfunction. 

The muscarinic M4 receptor has been studied as a target of interest for the treatment of 

schizophrenia; agonists or positive allosteric modulators (PAMs) are under consideration 

as potential antipsychotic agents. 

Recently, GPCR research has been enriched by the development of label-free technologies 

(using cellular impedance) which allow phenotypic measurement of cell function and are not 

restricted to single signalling pathways. Using this technology it is possible to evaluate 

whole cell responses to drug challenges and dissect kinetics of responses as well as biased 

agonism. 

Using this label-free technology, the pharmacology novel agonists and PAMs of the 

muscarinic M4 receptor will be studied; inhibitors of G protein coupling and signal 

transduction pathways will be used to characterise the functional responses; these will 

also be verified in orthogonal assays of muscarinic M4 receptor function. This will lead to 

a greater understanding of activation and allosteric modulation of the muscarinic M4 

receptor which will be of benefit in the development of novel antipsychotic drugs. 

Techniques: 

Pharmacological: Receptor function will be quantified by downstream signalling using stateof-the-art, 

high throughput assays of cellular impedance and possibly extracellular signal 

regulated kinases (ERK1/2), intracellular cAMP or resonance energy transfer techniques to 

detect interaction with β arrestin. 

Molecular biological: As part of the project you will also learn basic molecular biological 

procedures including isolation and preparation of DNA, cell culture and cell transfection. 

Contacts: 

Dr Chris Langmead 


chris.langmead@monash.edu 



School/Department.


INVESTIGATING THE SIGNALLING CASCADE OF NOVEL 

ADENOSINE RECEPTOR LIGANDS AT THE ADENOSINE A2B 

RECEPTOR (A2B-AR) 

Supervisors: Dr Celine Valant, Dr Lauren May 





Since its discovery as a low-affinity adenosine receptor (AR), the A2B receptor (A2B-AR), 

has proven enigmatic in its function. The lack of specific pharmacological agents targeting 

the A2B-AR made its initial characterization challenging. 

However, the importance of this receptor was reconsidered when it was observed that the 

A2B-AR is highly transcriptionally regulated by factors implicated in inflammatory 

hypoxia. The notion that during ischemia or inflammation extracellular adenosine is 

dramatically elevated to levels sufficient for A2B-AR activation, indicated that A2B-AR 

signaling may be important to dampen inflammation particularly during tissue hypoxia. 

Additionally, the involvement of this receptor in processes such as interleukins secretion, 

Ca2+ mobilization, hepatic glucose regulation, tumor vascularisation, and cardioprotection 

have stimulated many researchers to develop specific agonists and antagonists. For many 

years, the lack of potent and selective A2B-AR ligands precluded a deep exploration of 

their therapeutic prospective; at present, much progress in the field of antagonists led to 

preclinical studies for different compounds. Less populated is the universe of A2B-AR 

agonists, but really promising for the involvement in ischemic preconditioning. 

Here we propose to study the pharmacological properties of various adenosine ligands, 

including the recently in-house identified highly potent A2B-AR agonist, VCP746. Following 

receptor activation, the A2B-AR couple predominantly to Gs protein to induce cAMP 

production via stimulation of adenylate cyclases. As such cAMP production will be initially 

used as a standard assay to quantify agonist potency in the absence as well as in the 

presence of selective pathway inhibitors of various stages of the signalling cascade of the 

adenosine A2B-AR. Ultimately, if time permits, other signalling assays will be investigated, 

such as Ca2+ mobilization and ERK1/2 phosphorylation. 

Techniques: 

Cell culture: In this project you will learn the basics of cell culture, such as splitting and 

seeding cells. 

Pharmacology: Various singalling pathways will be learnt. Initially, cAMP production will be 

the main signaling assay investigated in this study but if the time permits, other 

downstream signalling can be investigated, such as ERK1/2 phosphorylation, calcium 

mobilization or cell survival. 

Contacts: 

Dr Celine Valant 

Phone: 9903 9091, Rm 344, Building 404; celine.valant@monash.edu 



School/Department.


INVESTIGATION OF AN ABSOLUTELY CONSERVED POCKET AT 

THE MUSCARINIC ACETYLCHOLINE M1 RECEPTOR (M1 MACHR) 

Supervisors: Dr Celine Valant, Dr Meritxel Canals 





Five muscarinic acetylcholine receptor (mAChR) subtypes have been cloned (M1, M2, M3, 

M4 and M5). For years now, there has been great interest in the structure-function 

relationships of mAChRs because these prototypical Family A G protein-coupled receptors 

(GPCRs) are attractive therapeutic targets for both peripheral and central nervous 

system disorders. A plethora of drugs that act at the mAChRs have been identified over 

the last few decades, but many of these show minimal selectivity for any one of the five 

mAChR subtypes over the others, which has hampered their development into 

therapeutics due to adverse side effects. The lack of drug specificity is primarily due to 

high sequence similarity in this family of receptor, especially in the orthosteric 

(endogenous ligand) binding pocket. 

Recently, with the success of crystallization of Family A GPCRs, especially in the mAChR 

family, a number of residues outside the endogenous ligand binding pocket, absolutely 

conserved cross all five mAChR subtypes, and highly conserved across the family A GPCRs, 

have been identified. These thirteen residues appear to form an extended binding pocket 

located right below the orthosteric binding site. In an effort to understand the purpose 

of such highly conserved deep pocket, we propose to investigate one by one the effect of 

each residue, using the M1 mAChR as a template. Starting with alanine mutation of each 

residue, the binding and functional properties of a selection of muscarinic ligands, 

antagonists, non-selective agonists, and especially ‘selective allosteric agonists’, such as 

77-LH-28, AC-42, TBPB and BQCA, will be investigated. 

Techniques: 


seeding cells, as well as cell transfection. 

Pharmacology: Radioligand binding assays as well as various signaling assays will be learnt 

during this study. 

Molecular biology: As part of the project you will also learn basic molecular biological 

procedures including isolation and preparation of DNA. 

Contacts: 



celine.valant@monash.edu 



School/Department.


INVESTIGATION OF AN ABSOLUTELY CONSERVED POCKET AT 

THE MUSCARINIC ACETYLCHOLINE M2 RECEPTOR (M2 MACHR) 

Supervisors: Dr Celine Valant, Prof Arthur Christopoulos 





Five muscarinic acetylcholine receptor (mAChR) subtypes have been cloned (M1, M2, M3, 

M4 and M5). For years now, there has been great interest in the structure-function 

relationships of mAChRs because these prototypical Family A G protein-coupled receptors 

(GPCRs) are attractive therapeutic targets for both peripheral and central nervous 

system disorders. A plethora of drugs that act at the mAChRs have been identified over 

the last few decades, but many of these show minimal selectivity for any one of the five 

mAChR subtypes over the others, which has hampered their development into 

therapeutics due to adverse side effects. The lack of drug specificity is primarily due to 

high sequence similarity in this family of receptor, especially in the orthosteric 

(endogenous ligand) binding pocket. 

Recently, with the success of crystallization of Family A GPCRs, especially in the mAChR 

family, a number of residues outside the endogenous ligand binding pocket, absolutely 

conserved cross all five mAChR subtypes, and highly conserved across the family A GPCRs, 

have been identified. These thirteen residues appear to form an extended binding pocket 

located right below the orthosteric binding site at the M2 mAChR. In an effort to 

understand the purpose of such highly conserved deep pocket, we propose to investigate 

one by one the effect of each residue. Starting with alanine mutation of each residue, the 

binding and functional properties of a selection of muscarinic ligands, such as antagonists, 

agonists and allosteric modulators will be thoroughly investigated. 

Techniques: 


seeding cells, as well as cell transfection. 

Pharmacology: Radioligand binding assays as well as various signaling assays will be learnt 

during this study. 

Molecular biology: As part of the project you will also learn basic molecular biological 

procedures including isolation and preparation of DNA. 

Contacts: 



celine.valant@monash.edu 



School/Department.


PROBING STRUCTURE ACTIVITY RELATIONSHIPS FOR 

GLUCAGON-LIKE PEPTIDE-1 RECEPTOR ALLOSTERIC LIGANDS 

Supervisors: Dr Denise Wootten and Prof Patrick Sexton 




Background: 

The glucagon-like peptide-1 (GLP-1) receptor (GLP-1R) is a family B GPCR that plays an 

essential role in nutrient mediated insulin release and as such is a therapeutic target for 

the treatment of type II diabetes. Unlike most GPCRs, the GLP-1R is activated by four 

distinct endogenous GLP-1 variants, as well as by further endogenous and exogenous 

ligands. Current GLP-1R based therapeutics for treatment of type II diabetes is 

restricted to daily intravenous or subcutaneous administrations of peptide analogues of 

GLP-1. Therefore, orally active, small molecule agonists of the GLP-1 receptor are highly 

desired. Recent developments in the GLP-1R field include the discovery of nonpeptide 

ligands, which bind at sites topographically distinct (allosteric) from the endogenous 

peptides. Allosteric ligands offer alternative routes to receptor activation that may be 

able to either modulate the existing peptide response and/or activate the receptor alone. 

We have recently characterised two of these nonpeptide GLP-1R allosteric ligands, 

Compound 2 and BETP. In recombinant cell systems, Compound 2 was a strong partial 

agonist in cAMP signalling and a positive allosteric modulator of oxyntomdulin activity (but 

not of other orthosteric agonists), whereas BETP displayed only weak partial intrinsic 

agonsim in cAMP, but similarly to Compound 2, selectively modulated oxyntomodulin 

affinity and efficacy in cAMP signalling. 

Project aim: 

The aim of this project is to screen in a library of compounds based on the scaffolds of 

Compound 2 and BETP in the classical signalling pathway (cAMP) to identify a structureactivity 

profile for intrinsic efficacy and modulator activity. In addition, other signalling 

pathways (such as pERK1/2, iCa2+ and b-arrestin recruitment) will be assessed in an 

attempt to discover biased and/or probe-dependent ligands which may be used in future 

studies to interrogate the in vivo biology of the GLP-1R. 

Techniques: 

Receptor function will be quantified by downstream signalling using state-of-theart, 

high throughput assays of intracellular cAMP, extracellular signal regulated 

kinases (ERK1/2), iCa2+ mobilization assays and resonance energy transfer 

techniques to detect interaction with β arrestins. Radioligand binding assays will be 

used to assess ligand affinities. As part of the project you will also learn various 

cell culture techniques. 

Contacts: Dr Denise Wootten 

Phone: 9903 9088, Rm 5-09, Building 404; Denise.wootten@monash.edu 



School/Department.

Current Honours Projects - Faculty of Medicine, Nursing and Health ...

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