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New Horizons 2018 | Innovative Science with Impact

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FINAL PROGRAM & ABSTRACT BOOK

Kolling Research Institute 19th – 20th November 2018

35th Combined Health Science Conference

New Horizons 2018 Innovative science with impact:

Strengthening alliances between research

New Horizons 2018 | Innovative science with impact

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ACKNOWLEDGEMENTS

The New Horizons 2018 Program Committee would like to thank the following who, at the time of printing, had given their support to this conference: INSTITUTIONS GOLD SPONSOR SILVER SPONSOR

EXHIBITORS & OTHER SPONSORS (in alphabetical order)


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TABLE OF CONTENTS

WELCOME .......................................................................................................................................... 3

PROGRAM AT A GLANCE .................................................................................................................... 4

ORGANISING COMMITTEE ................................................................................................................. 6

INVITED SPEAKERS ............................................................................................................................. 7

DETAILED PROGRAM DAY 1 (Monday 19th Nov).............................................................................. 12

DETAILED PROGRAM DAY 2 (Tuesday 20th Nov) ............................................................................ 14

ORAL PRESENTATION ABSTRACTS ................................................................................................... 15

POSTER PRESENTATION ABSTRACTS ............................................................................................... 15

AWARDS AND PRIZES ....................................................................................................................... 15

GENERAL INFORMATION ................................................................................................................. 15

KOLLING INSTITUTE FLOOR PLANS .................................................................................................. 15

SPONSOR AND EXHIBITOR INFORMATION ...................................................................................... 15


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WELCOME

On behalf of the Organising Committee and all co-hosting organisations I would like to extend a very warm welcome to delegates, plenary speakers, presenters and invited guests of New Horizons 2018. This is the 35th Combined Health Science Conference co-hosted by the Kolling Institute, the University of Sydney, Northern Sydney Local Health District, the University of Technology Sydney and the Centenary Institute. Through this meeting, we look forward to forming new collaborations and strengthening the continuous relationship between our Institutions.

These annual conferences promote advancements in healthcare through research and education, and by building bridges between basic science, the clinic and delivery of care. There is a rich history of collaboration between the institutes involved which, over the past 34 years, has resulted in significant progress in all of these areas. This year our conference will focus on research with impact - research that increases our understanding of disease and leads to the development of new therapies and better outcomes for patients. It will include presentations on the latest scientific research, new developments in technology, clinical excellence and the translation of research from bench to bedside. New Horizons is held at the Kolling Institute, located within the Royal North Shore Hospital. This strategic location enables the clinical translation of research outcomes and the development of research with impact. We are very grateful to our plenary and invited speakers for agreeing to share their latest research with us. We also look forward to presentations and posters from health care providers, research assistants, Honours students, PhD students, Masters students, post-doctoral researchers as well as research leaders. We acknowledge and sincerely thank all of those who have committed time and effort into the organising of this conference, our academic and health sponsors, as well as our commercial sponsors, who have made the meeting possible. A special thanks to our Gold sponsors, Spruson and Ferguson, for continuously supporting this Meeting. We hope that you find this meeting productive and informative, and take the opportunity to form new cross-institutional and inter-disciplinary collaborative and personal relationships. We have tried to create a relaxed and friendly atmosphere that allows students and younger researchers to feel comfortable presenting their research and interacting with more experienced researchers.

Please join us for drinks and canapés after the meeting - take the opportunity to interact with the speakers and other delegates and enjoy being part of a vibrant research community. Dr Sonia Saad, Organising Committee Chair, 2018

DR SONIA SAAD Adjunct Professor, University of Technology Sydney Senior Research Fellow Medicine, Northern Clinical School, University of Sydney

Invited speakers


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PROGRAM AT A GLANCE

MONDAY 19th NOVEMBER 2018

8 am – 5 pm Registration desk open

9:15 – 9:30 am Welcome and Introduction to Conference Prof Carolyn Sue (Kolling Institute Executive Director)

9:30 – 10:20 am Opening Plenary Talk Prof Hala Zreiqat (University of Sydney, Sydney, NSW) Novel synthetic approaches to Innovative BioEngineering

10:20 – 10:40 am Morning tea, exhibitions

10:40 – 11:20 am Plenary Session 1: Infection and immunity Prof Michael Hickey (Monash University, Melbourne, VIC) Imaging initiation of T cell-mediated glomerulonephritis

11:20 – 11:30 am Silver sponsor

11:30 – 12:10 pm Abstract Session 1: Honours students and Research Assistants’ Talks (5 mins each)

12:10– 12:30 pm Guided Poster Session

12:30 – 1:30 pm Lunch, exhibitions

1:30 – 2:10 pm Plenary Session 2: Cancer Epigenetics Prof Susan Clark (Garvan Research Institute, Sydney, NSW) New Insights into the Cancer Epigenome

2:10 – 3:00 pm Abstract Session 2: Early Career Researchers’ Talks I (10 mins each)

2:50 – 3:20 pm Afternoon tea, posters, exhibitions

3:20 – 4:00 pm Plenary Session 3: The microbiome in health and disease Prof Emad El-Omar (St George & Sutherland Clinical School, Kogarah, NSW) The human microbiome: where are we now?

4:00 – 4:40 pm

Q & A session on Translation and Commercialisation Panel members: Dr Alison Todd (SpeedX, Sydney, NSW) Dr Leszek Lisowski (CMRI /LogicBio Therapeutics, Sydney, NSW) Ms Kathy Connell (J&J Innovation, Sydney NSW) Chair: Prof Michael Wallach (UTS, Sydney, NSW)

4:40 – 6:00 pm ECR session with drinks


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TUESDAY 20th NOVEMBER 2018

8 am – 5 pm Registration desk open

8:50 – 9:20 am

Plenary Session 4: Medical Research Rising Stars (inaugural session) Dr Ben Roediger (Centenary Institute, Sydney, NSW) An atypical parvovirus drives chronic tubulointerstitial nephropathy and kidney fibrosis

9:20 – 10:05 am Abstract Session 3: Higher Degree Research Students’ Talks I (7 mins each)


10:35 – 11:15 am Plenary Session 5: Cardiovascular Diseases and Endocrinology Dr John O’Sullivan (Heart Research Institute, Sydney, NSW) Refining cardiometabolic disease

11:15 – 11:25 am MediKane

11:25 – 12:15 pm Abstract Session 4: Higher Degree Research Students’ Talks II (7 mins each)


1:00 – 1:40 pm Plenary Session 6: Neurobiology Prof Emeritus Alan Mackay Sim (Griffith University, Brisbane, QLD) Stem cell models of brain diseases

1:40 – 1:55 pm Gold sponsor

1:55 – 2:45 pm Abstract Session 5: Early Career Researchers’ Talks II (10 mins each)

2:45 – 3:10 pm Afternoon tea, Exhibitions

3:10 – 3:30 pm Guided Poster Session

3:30 – 4:10 pm

Session 7: Pharmacology Prof Renae Ryan (University of Sydney, NSW) The Split Personality of Glutamate Transporters: a Chloride Channel and a Transporter

4:10 – 4:30 pm Closing Remarks and Awarding of Prizes

Prof Alaina Ammit (UTS Associate Dean Research) Prof Carolyn Sue (Kolling Institute Executive Director) Prof Mathew Vadas (Centenary Institute Executive Director)

4:30 – 5:30 pm Canapés Reception

Invited speakers


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ORGANISING COMMITTEE The Organising Committee of the New Horizons 2018 Conference includes (in alphabetical order):

• A/Prof Beata Bajorek, Graduate School of Health, UTS and Academic Pharmacist, Royal North

Shore Hospital, NSLHD

• A/Prof Patrick Bertolino: Faculty and Head of the Liver Immunology group, Centenary Institute

• Dr Catherine Burke: Lecturer, School of Life Sciences, UTS

• Dr Valery Combes, Senior Lecturer, School of Life Sciences, UTS

• Dr Alvaro Garcia: Chancellor's Postdoctoral Research Fellow, School of Life Sciences, UTS

• Dr Cindy Gunawan: Senior Lecturer, The ithree Institute, Core Member, ithree - Institute of Infection, Immunity and Innovation, UTS

• Dr Jiao Jiao Li: NHMRC Peter Doherty Biomedical Early Career Fellow, Kolling Institute of Medical Research Northern Clinical School

• Dr Kristine McGrath, Lecturer, School of Life Sciences, UTS

• Dr. Sonia Saad, Senior Research Fellow, Kolling Institute and Adjunct Professor, School of Life Sciences, UTS

• Dr Jessamy Tiffen: Research Officer, Melanoma Immunology and Oncology Program, Centenary Institute

• Dr Nham Tran: Deputy Head of School and Principal Investigator, School of Biomedical Engineering, UTS


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INVITED SPEAKERS Invited Speakers of the New Horizons 2018 Conference (in alphabetical order):

Prof Hala Zreiqat Hala Zreiqat is is a professor of biomedical engineering at the University of Sydney and both a National Health and Medical Research Council Senior Research Fellow (2016-2020). She is the 2018 New South Wales Premier’s Woman of the Year. Her research is on the development of novel engineered materials and 3D-printed platforms for regenerative medicine, particularly in orthopaedic, dental, and maxillofacial applications. Prof Zreiqat’s overall objective is to advance collaborative research ventures and build educational and industry linkages nationally and internationally in the field of musculoskeletal disorders and biomaterials research. Prof Michael Hickey Michael Hickey is the Head of the Leucocyte trafficking group at Monash University. He completed his PhD at the University of Melbourne in 1996. He then underwent postdoctoral training in the Immunology Research Group at the University of Calgary in Canada (1996-1999) and the Baker Heart Research Institute (2000-2001), before joining the Centre for Inflammatory Diseases at Monash University in August 2001. His laboratory investigates the behaviour of leukocytes in inflamed tissues and the blood vessels that supply them, using highly advanced microscopes to visualise white blood cells in tissues. The aim of his research is to understand how they contribute to inflammatory diseases, and thereby identify new ways to tackle these conditions. Prof Susan Clark Susan Clark is a NHMRC Senior Principal Research Fellow and Head of the Genome and Epigenetics Division at the Garvan Institute of Medical Research in Sydney, Australia. She graduated in 1982 with a PhD in Biochemistry, University of Adelaide. Her molecular studies over her career have addressed profound questions about the importance of epigenetics in early development and in disease, especially in cancer. The techniques she pioneered in the early 1990s, including bisulphite methylation sequencing, helped to revolutionise epigenomic research. Susan was a founding member of IHEC (International Human Epigenome Consortium) and led the formation of the AEpiA (Australian Epigenetics Alliance).

Professor Emad El-Omar Emad El-Omar graduated in Medicine from Glasgow University, Scotland, and trained as a gastroenterologist. He worked as a Visiting Scholar/Scientist at Vanderbilt University, TN, and National Cancer Institute, MD, USA, and was Professor of Gastroenterology at Aberdeen University, Scotland, for 16 years before taking up the Chair of Medicine at St George & Sutherland Clinical School, University of New South Wales, Sydney, Australia. He is the Editor in Chief of the journal Gut. His research interests include the gut microbiome, inflammation driven GI cancer and IBD. He is the Director of the Microbiome Research Centre at St George Hospital, Sydney. Dr Alison Todd Alison Todd is Chief Scientific Officer at SpeeDx. She has extensive experience within the pharmaceutical industry where she developed several novel molecular analytical technologies which have been used for basic research, preclinical/clinical drug development and in vitro diagnostics. Her specific areas of scientific expertise include nucleic acid chemistry, particularly target amplification and catalytic DNA technologies, and the biology of cancer and viral diseases. Alison is an inventor on 18 patents/patent applications and her techniques have been used for molecular monitoring in nine international clinical trials for patients with HIV, leukaemia and various solid tumours. She has successfully championed multiple inventions through to product commercialization in various fields.

Invited speakers


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Dr Leszek Lisowski Leszek Lisowski is a world vectorology expert with over 15 years of experience in developing and manufacturing viral vectors for human gene therapy. He was born in a small town in Western Poland. After high school, he received a prestigious Academic Excellence and Leadership Scholarship and enrolled at the University of Bridgeport, Bridgeport, CT, USA where he earned his B.S. in Biology. He subsequently joined a doctorate program at Cornell University in New York. He earned his PhD in Molecular Biology and Genetics from the laboratory of Prof Michel Sadelain, MD PhD at the Memorial Sloan Kettering Cancer Center (MSKCC), for his work on lentiviral vectors for the treatment of β-thalassemia and assessment of vector toxicity and the risk of insertional oncogenesis. Ms Kathy Connell Kathy Connell is Senior Director of New Ventures, Australia and New Zealand (ANZ) for Johnson & Johnson Innovation, California and is responsible for implementing our strategy across Australia and New Zealand. She has a clinical background and started her commercial career as the VP of Global Business Development for an Australian biotechnology company. Prior to joining Janssen, Kathy worked for Sanofi engaged in in-licensing late stage pharmaceutical assets. Kathy is a registered psychologist and holds Post Graduate Diplomas in Health and Medical Law from the University of Melbourne, Psychology from Monash University and Bachelor degrees in Psychology (Swinburne University) and Applied Science from LaTrobe University Dr Ben Roediger Dr Roediger is the Head of the Skin Imaging and Inflammation Laboratory at the Centenary Institute. He was awarded his PhD in 2011. His research combines cutting-edge technology with sophisticated mouse models to visualise and functionally characterise immune cells in vivo. Amongst his key achievements, he provided the first ever identification of skin ILC2 and demonstrated that uncontrolled activation and expansion of these cells is sufficient to induce inflammatory skin disease. Dr Roediger was also involved in the first ever visualisation of the innate immune response to Staphylococcus aureus infection in vivo, which identified a hitherto unrecognised role for perivascular macrophages in neutrophil recruitment (Abtin et al. Nat. Immunol. 2014).

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Dr John O’Sullivan John O’Sullivan is a Clinical-Academic Cardiologist at the Royal Prince Alfred Hospital and Group Leader in Cardiometabolic Disease at the Heart Research Institute and Charles Perkins Centre of the University of Sydney. John undertook his internal medicine, cardiology, and PhD training in Ireland, and then spent 4 years at Massachusetts General Hospital and Harvard Medical School. John studies the cardiovascular consequences of obesity and related diseases such as diabetes. He is particularly interested in applying unbiased non-targeted metabolomic profiling to discover novel disease insight. In carefully-designed patient cohorts, he leverages metabolomic and genomic data to determine novel markers, effectors, and predictors of disease; he then interrogates their functional roles in model systems.

Prof Emeritus Alan Mackay Sim Alan Mackay-Sim is Professor Emeritus at Griffith Institute for Drug Discovery, Griffith University, Brisbane, Australia. He was named 2017 Australian of the Year in public recognition of his research on how the olfactory sensory neurons in the nose get regenerated throughout life. He has identified the olfactory stem cell in the nose that is responsible for the regeneration of the sense of smell and uses these “adult” stem cells and other olfactory cells from the nose for therapeutic purposes. To this end he established the National Centre for Adult Stem Cell Research in 2006 and built a unique resource: a bank of neural stem cells from over 300 people, healthy controls and from patients with neurological conditions including schizophrenia, Parkinson’s disease, mitochondrial mutation disorders, Hereditary

Invited speakers


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Spastic Paraplegia, ataxia telangiectasia and motor neuron disease. These are used for understanding the biological bases of neurological diseases and for drug discovery.

Prof Renae Ryan Renae Ryan is Professor of Biochemical Pharmacology at the University of Sydney and the Academic Director of SAGE. She received her PhD from the University of Sydney in 2004. After working as a postdoctoral fellow at Columbia University and the National Institutes of Health in the USA, Renae returned to the University of Sydney and was appointed as Associate Professor in the Sydney Medical School in 2010. She leads a research team that investigates molecular pumps that transport amino acids into cells and designs novel compounds that target these pumps to treat diseases such as chronic pain and cancer. Renae is Chair of the Gender Equity Committee and the Early Career Researcher Network in the Sydney Medical School. She has received several prestigious awards and fellowships including a NSW Tall Poppy Award and a NHMRC Career Development Fellowship.


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DETAILED PROGRAM DAY 1 (Monday 19th Nov) 8 am – 5 pm Registration desk open

9:15 – 9:30 am Welcome and Introduction to Conference Prof Carolyn Sue (Kolling Institute Executive Director)

9:30 – 10:20 am Opening Plenary Talk Prof Hala Zreiqat (University of Sydney, Sydney, NSW) Novel synthetic approaches to Innovative BioEngineering Chair: Prof Christopher Little


10:40 – 11:20 am Plenary Session 1: Infection and immunity Prof Michael Hickey (Monash University, Melbourne, VIC) Imaging initiation of T cell-mediated glomerulonephritis Chair: Dr Muh-Geot Wong

11:20 – 11:30 am Silver sponsor

11:30 – 12:10 pm

11:30 am

11:35 am

11:40 am

11:45 am

11:50 am

11:55 am

12:00 pm

12:05 pm

Abstract Session 1: Honours students and Research Assistants’ Talks (5 mins each) Chair: A/Prof Usha Panchapakesan, Co-chair: Dr Alvaro Garcia Abstract 1-1: Karen Schuck (The University of Sydney) Optimising primary care management of knee osteoarthritis (the partner study): lessons from a non-randomised pilot study

Abstract 1-2: Stephanie Sun (Centenary Institute) N6-methyladenosine (M6A) regulates terminal granulocyte differentiation

Abstract 1-3: Mawj Mandwie (UTS) Metformin treatment ameliorates high-fat diet Induced neuroinflammation in the mouse pre-frontal cortex and hippocampus –implications for the PACAP/VIP system

Abstract 1-4: Jocelyn Karinua (UTS) PACAP and VIP modulate lipopolysaccharide (LPS)-induced microglial activation by triggering distinct phenotypic changes

Abstract 1-5: Khalia Ackermann (Centenary Institute) The response of CD8 T cells recognising liver-expressed antigens is regulated by the affinity of their TCR for p:MHC ligands, favouring high affinity clones

Abstract 1-6: Shihani Stoner (Raymond Purves Bone and Joint Research Labs) Arc (apoptosis repressor with caspase recruitment domain) deficient mice develop severe post-traumatic knee osteoarthritis but have reduced pain

Abstract 1-7: Natalia Pinello (Centenary Institute) Elucidating the role of overexpressed N 6-methyladenosine methyltransferase,VIRMA, in breast cancer

Abstract 1-8: Stuart Cook (Centenary Institute) Differential chemokine receptor expression and usage by immediate dendritic cell precursors



1:30 – 2:10 pm Plenary Session 2: Cancer Epigenetics Prof Susan Clark (Garvan Research Institute, Sydney, NSW) New Insights into the Cancer Epigenome

Chair: Ass. Prof Roderick Clifton-Bligh


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DETAILED PROGRAM DAY 1 (Monday 19th Nov) 2:10 – 3:00 pm

2:10 pm

2:20 pm

2:30 pm

2:40 pm

Abstract Session 2: Early Career Researchers’ Talks I (10 mins each)

Chair: Dr Jessamy Tiffen, Co-chair: Dr Ryan Davis

Abstract 1-9: Dr Alvaro Garcia (UTS) Modulation of cation interactions with zwitterionic phospholipid bilayers by headgroup associated carbonyl oxygens. Abstract 1-10: Dr Long The Nguyen (Kolling Institute) Tissue-specific roles of SIRT-1 in the developmental origin of metabolic disorders due to maternal high-fat feeding Abstract 1-11: Dr Elinor Hortle (Centenary Institute) Inhibition of thrombocyte activation restores protective immunity to mycobacterial infection Abstract 1-12: Dr Lisa Kouladjian O’Donnell (Kolling Institute) Implementation of the goal-directed medication review electronic decision support system (GMEDSS) into home medicines review (HMR) to deprescribe medications in older adults

2:50 – 3:20 pm Afternoon tea, posters, exhibitions

3:20 – 4:00 pm Plenary Session 3: The microbiome in health and disease Prof Emad El-Omar (St George & Sutherland Clinical School, Kogarah, NSW) The human microbiome: where are we now? Chair: Prof Christopher Jackson

4:00 – 4:40 pm

Q & A session on Translation and Commercialisation Panel members: Dr Alison Todd (SpeedX, Sydney, NSW) Dr Leszek Lisowski (CMRI /LogicBio Therapeutics, Sydney, NSW) Ms Kathy Connell (J&J Innovation, Sydney NSW)

Chair: Prof Michael Wallach (UTS, Sydney, NSW)

4:40 – 6:00 pm ECR session with drinks


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DETAILED PROGRAM DAY 2 (Tuesday 20th Nov) 8 am – 5 pm Registration desk open

8:50 – 9:20am

Plenary Session 4: Medical Research Rising Stars (inaugural session) Dr Ben Roediger (Centenary Institute, Sydney, NSW) An atypical parvovirus drives chronic tubulointerstitial nephropathy and kidney fibrosis Chair: Ass. Prof Patrick Bertolino

9:20 – 10:05 am

9:20 am

9:27 am

9:34 am

9:41 am

9:48am

9:55 am

Abstract Session 3: Higher Degree Research Students’ Talks I (7 mins each) Chair: Dr Emily Colvin, Co-chair: Dr Jiao Jiao Li

Abstract 2-1: Gerard Li (UTS) Intrauterine cigarette smoke exposure increases metabolic and liver damage which were reversed by mitochondrial antioxidant

Abstract 2-2: Gizem Gemikonakli (Kolling Institute) Effect of chronic polypharmacy and the Drug Burden Index (DBI) on muscle function and structure in aged mice.

Abstract 2-3: Imanuelle Green (Centenary Institute) Macrophage development and activation involves coordinated intron retention in key inflammatory regulators

Abstract 2-4: Carina Blaker (Kolling Institute) Risk of ACL rupture and osteoarthritis: a role for prior sub-critical injury

Abstract 2-5: Benjamin Sealy (UTS) The role of extracellular vesicles in the modulation of endothelial junctions in an in vitro model of cerebral malaria

Abstract 2-6: Fathima Shihana (The University of Sydney) Chronic alcohol drinking alters microRNA in serum in patients with liver cirrhosis


10:35 – 11:15 am Plenary Session 5: Cardiovascular Diseases and Endocrinology Dr John O’Sullivan (Heart Research Institute, Sydney, NSW) Refining cardiometabolic disease Chair: Prof Christoper Ward

11:15 – 11:25 am MediKane

11:25 – 12:15 pm

11:25 am

11:32 am

11:39 am

11:46 am

11:53 am

12:00 pm

12:07 pm

Abstract Session 4: Higher Degree Research Students’ Talks II (7 mins each) Chair: Dr Giles Best, Co-chair: Dr Piklu Roy Chowdhury Abstract 2-8: Jessica Pedersen (UTS) miR-99b and miR-652 are required for optimal control of M. Tuberculosis infection in human macrophages

Abstract 2-9: Alex Wong (Centenary Institute) Prpf19 regulates intron retention in acute myeloid leukaemia

Abstract 2-10: Yeon Jae Kim (Kolling Institute) Bringing back the old – Ouabain a promising treatment for new-onset atrial fibrillation?

Abstract 2-11: Riti Mann (UTS) We are what we eat: identifying a regulatory crosstalk between central carbon metabolism and cell division in bacteria Abstract 2-12: Kieran English (Centenary Institute) CD4 T cell help is required for robust CD8 T cell to hepatocyte-expressed antigens

Abstract 2-13: Qinghua Cao (Kolling Institute and Royal North Shore Hospital) A novel therapeutic strategy for kidney fibrosis with a small protein I-body (AD-114)

Abstract 2-14: Kathy On (Centenary Institute) The role of perivascular macrophages in contact hypersensitivity


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DETAILED PROGRAM DAY 2 (Tuesday 20th Nov) 12:15 – 1:00 pm Lunch, exhibitions

1:00 – 1:40 pm Plenary Session 6: Neurobiology Prof Emeritus Alan Mackay Sim (Griffith University, Brisbane, QLD) Stem cell models of brain diseases Chair: Prof Carolyn Sue

1:40 – 1:55 pm Gold sponsor: Spruson & Ferguson IP and Commercialisation of Research

1:55 – 2:45 pm

1:55 pm

2:05 pm

2:15 pm

2:25 pm

2:35 pm

Abstract Session 5: Early Career Researchers’ Talks II (10 mins each) Chair: Dr Sharon McCracken, Co-chair: Dr Martin Bullock

Abstract 2-15: Dr Yingyu Feng (The University of Sydney) Patterns of immediate breast reconstruction in NSW Australia: a population-based study

Abstract 2-16: Dr Jiao-Jiao Li (Ray Purves Bone and Joint Research Labs & Kolling Institute) Cross-talk between stem cells and diseased cells in an osteoarthritic joint: implications for treatment

Abstract 2-17: Dr Sioh-Yang Tan (Centenary Institute) The role of liver capsular macrophages in hepatic fibrosis

Abstract 2-18: Dr Ryan Davis (Kolling Institute) Whole Genome Sequencing and intronic sequence interrogation to solve a case of early-onset familial Parkinson’s Disease

Abstract 2-19: Dr Lana McClements (UTS, Sydney, NSW) FKBPL, a novel player in heart ischaemia and fibrosis

2:45 – 3:10 pm Afternoon tea, Exhibitions


3:30 – 4:10 pm

Plenary Session 7: Pharmacology Prof Renae Ryan (University of Sydney, NSW) The Split Personality of Glutamate Transporters: a Chloride Channel and a Transporter Chair: Prof Sarah Hilmer

4:10 – 4:30 pm Closing Remarks and Awarding of Prizes

Prof Alaina Ammit (UTS Associate Dean Research) Prof Carolyn Sue (Kolling Institute Executive Director) Prof Mathew Vadas (Centenary Institute Executive Director)

4:30 – 5:30 pm Canapés Reception


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ORAL PRESENTATION ABSTRACTS

Speaker abstracts are provided in presentation order (presenting author underlined): ` MONDAY 19TH NOVEMBER 2018

Abstract 1.1 TITLE: OPTIMISING PRIMARY CARE MANAGEMENT OF KNEE OSTEOARTHRITIS (THE PARTNER STUDY): LESSONS FROM A NON-RANDOMISED PILOT STUDY Karen Schuck1, Jocelyn L Bowden1 , Charlotte Marshall3, Michelle King8, Kim L Bennell3, Rana S Hinman3, Thorlene Egerton3, Andrew M Briggs4, Stephen J Bunker5,11, Andrew B Forbes6, Simon D French7, Jessica Kasza6, Marie Pirotta8, Deborah J Schofield9, Nick A Zwar10, David J Hunter1,2. 1 Institute of Bone and Joint Research, Kolling Institute, University of Sydney, Sydney, NSW, Australia. 2 Rheumatology Department, Royal North Shore Hospital, Sydney, NSW, Australia. 3 Centre for Health, Exercise and Sports Medicine, Department of Physiotherapy, The University of Melbourne, Melbourne, VIC, Australia.

4 School of Physiotherapy and Exercise Science, Curtin University, Perth, WA, Australia.

5 Medibank, Docklands, VIC, Australia. 6 Biostatistics Unit, School of Public Health and Preventive Medicine, Monash University, Melbourne, VIC, Australia. 7Department of Chiropractic, Faculty of Science and Engineering, Macquarie University, NSW, Australia. 8 Department of General Practice, The University of Melbourne, Melbourne, VIC, Australia.

9 Department of Economics, Faculty of Business and Economics, Macquarie University, NSW, Australia. 10 School of Public Health and Community Medicine, UNSW, Sydney, NSW, Australia.

11 School of Public Health and Preventive Medicine, Monash University, Melbourne, VIC, Australia. Background: The PARTNER study is evaluating a new model of service delivery to increase uptake of key non-surgical clinical recommendations for osteoarthritis (OA) management. Aim & Objectives: Test key components of the randomised controlled trial protocol and identify issues affecting wider implementation. Methods: Two general practices, their GPs (n=8) and patients (n=12) were recruited and received the intervention as described in the protocol (May-Dec 2017). There was no control group. GPs were offered training on current best-practice OA management. Patients received an OA diagnosis and referred to a centralised, multidisciplinary care support team (CST), trained in evidence-based OA management and behaviour change support. Patients received tailored educational materials, a strengthening program, and access to a weight-loss program if appropriate. Four main study components were examined: key processes; delivery; study procedure implementation; and scientific quality. Results: A mixed methods process evaluation was undertaken. At the end of their participation, all patients interviewed (n=9) rated their participation as “simple and straightforward”. Eight reported being satisfied with the CST support, and having a better understanding of their OA and self-management options. Completion of GP training was our most significant issue. Feedback from GPs suggested a combination of time constraints (too long), limitations with our online-delivery platform, and some redundancy in our content. Delivery of GP training has been restructured for the main trial. Conclusion: We incorporated all changes identified into the main trial protocol. PARTNER will fill a major evidence-to-practice gap in primary care management of OA, a major Australian public health burden.

ABSTRACT SESSION 1: HONOURS STUDENTS AND RESEARCH ASSISTANTS’ TALKS


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.2 N6-METHYLADENOSINE (M6A) REGULATES TERMINAL GRANULOCYTE DIFFERENTIATION Stephanie Sun1,2, Chau To Kwok1,2, Renhua Song1,2, John EJ Rasko2,3,4, Justin J-L Wong1,2

1 Epigenetics and RNA Biology Program, Centenary Institute, University of Sydney, Camperdown 2050, NSW, Australia 2 Sydney Medical School, University of Sydney, Camperdown 2050, NSW, Australia 3 Gene and Stem Cell Therapy Program, Centenary Institute, University of Sydney, Camperdown 2050, NSW, Australia 4 Cell and Molecular Therapies, Royal Prince Alfred Hospital, Camperdown 2050, NSW, Australia Background: Like DNA and protein modifications, RNA can similarly be reversibly modified. Of the 170 known reversible RNA modifications, N6-methyladenosine (m6A) is the most abundant and is responsible for regulating gene expression by altering RNA splicing, export and degradation. With functional importance in all cellular process, the role of m6A in physiological and aberrant granulocytic differentiation remains unknown. Aim & Objectives: We determine whether m6A functions in normal and aberrant granulocyte differentiation in vivo and in HL-60, an in vitro leukemia cell line. Methods: Using methylated RNA immunoprecipitation sequencing, we determined differential m6A-methylated mRNAs in terminally-differentiated mouse granulocytes compared to their progenitors. Differential m6A events were functionally characterised using Gene Ontology (GO) analysis to identify m6A-regulated biological processes. Lentiviral transduction with shRNAs targeting an enzyme responsible for depositing m6A, METTL3, was used to examine changes in all-trans retinoic acid (ATRA)-induced granulocyte differentiation. Results: 401 differential m6A events from 359 genes were observed between primary granulocytes and their progenitors. Differentially m6A-modified genes were enriched for functions including transcription, RNA binding and cell structure. These genes include hnRNPA1, a RNA binding protein, which has previously been demonstrated to inhibit expression of C/EBP, a key transcription factor in granulocyte differentiation. Knockdown of METTL3 in HL-60 induces auto-differentiation and sensitises cells to ATRA-mediated granulocyte maturation. Conclusion: m6A is functionally important in granulopoiesis by modifying genes involved in the differentiation of myeloid progenitors into terminal granulocytes. Furthermore, reducing m6A levels can overcome differentiation blocks in leukemic cells in vitro. Abstract 1.3 METFORMIN TREATMENT AMELIORATES HIGH-FAT DIET INDUCED NEUROINFLAMMATION IN THE MOUSE PRE-FRONTAL CORTEX AND HIPPOCAMPUS – IMPLICATIONS FOR THE PACAP/VIP SYSTEM Mawj Mandwie1, Jocelyn Karunia1, Aram Niaz1, Ghaith Al-Badri1, Claire Rennie1, Kristine McGrath1, Alessandro Castorina1,2

1 School of Life Sciences, Faculty of Science University of Technology Sydney, NSW, Australia 2 Discipline of Anatomy & Histology, School of Medical Science, The University of Sydney, NSW, Australia Background: Chronic high fat diet (HFD)/Obesity is thought to induce neuroinflammation and consequent cognitive and behavioural impairments. Metformin, a first-line medication for the treatment of type 2 diabetes, has shown promise in ameliorating cognitive and behavioural dysfunctions in neurodegenerative diseases, but the mechanism(s) are not clear yet. Aim & Objectives: We aimed at assessing whether HFD could lead to neuroinflammation in vulnerable brain regions and if it caused dysregulations in the natural neuroprotective/anti-inflammatory Pituitary Adenylate Cyclase-Activating Peptide/Vasoactive-Intestinal-Polypeptide (PACAP/VIP) system. Additionally, we tested if metformin treatment ameliorated HFD-brain inflammation and rescued the disruptions in this system through activating the neuroprotective PI3K/Akt pathway. Methods: Pre-frontal cortices and hippocampi from C57BL/6J male mice fed either with a Standard Chow, HFD or HFD followed by Metformin administration were micro-dissected. The inflammatory profile, expression of the PACAP/VIP system members and the activity of the PI3K/Akt pathway were assessed in the selected regions by real-time quantitative polymerase chain reaction (qPCR) and Western blotting. Results: Our data show that HFD causes inflammation both in the pre-frontal cortex and the hippocampus, which is associated with disruptions of the PACAP/VIP system and reductions of PI3K/Akt activity. Intriguingly, metformin treatment attenuated neuroinflammation and PACAP/VIP system disruptions caused by HFD, and rescued the PI3K/Akt cascade. Conclusion: Our findings suggest that HFD triggers inflammation in vulnerable brain regions, likely by disrupting the endogenous levels of PACAP/VIP peptides and downstream PI3K/Akt pathways. Metformin fully rescues HFD-induced detrimental effects. Our findings provide evidence to repurpose Metformin as a therapeutic option to treat brain-inflammatory disorders.


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.4 PACAP AND VIP MODULATE LIPOPOLYSACCHARIDE (LPS)-INDUCED MICROGLIAL ACTIVATION BY TRIGGERING DISTINCT PHENOTYPIC CHANGES Jocelyn Karunia1, Aram Niaz1, Mawj Mandwie1, Ghaith Al-Badri1, Alessandro Castorina1,2

1 School of Life Sciences, University of Technology Sydney, NSW, Australia 2 Discipline of Anatomy & Histology, School of Medical Science, The University of Sydney, NSW, Australia Background: Aberrant microglial activation plays a key role in the progressive neuronal loss seen in many neurodegenerative diseases. PACAP and VIP are two neuropeptides with robust immunosuppressive properties within the CNS. However, not much is known on the underlying mechanisms through which the peptides regulate microglia activities. Aim & Objectives: Here, using lipopolysaccharide (LPS) to induce microglial cell activation, we aimed at testing whether and how administration of either PACAP or VIP could differentially affect microglial pro-inflammatory profile, activation state and morphological appearance to produce immunosuppressive effects. Methods: Quantitative real-time PCR, Western blot, Griess reactions, immunofluorescence and morphological analyses were conducted in order to determine the effects of PACAP and VIP in BV2 microglial cells exposed or not to 1µg/ml LPS. Results: Our results showed that both PACAP and VIP reduce the expression of pro-inflammatory mediators on LPS-stimulated BV2 cells. We also found that exogenous administration of peptides rescued the dysregulations of the endogenous PACAP/VIP levels and attenuated the expression of microglial activation markers caused by LPS. Interestingly, we noted that, despite PACAP and VIP elicited similar anti-inflammatory activities in LPS-treated BV2 cells, their produced distinct effects on cell morphology, with PACAP mainly reducing the number of activated cells, whereas VIP acted by increasing the subpopulation of cells exhibiting an ‘intermediate’ phenotype/bipolar-shaped (p<0.001 vs. control), at the expenses of resting/rounded cells.

Abstract 1.5 The response of CD8 T cells recognising liver-expressed antigens is regulated by the affinity of their TCR for p:MHC ligands, favouring high affinity clones Khalia Ackermann1, Kieran English1, Sioh-Yang Tan1, Leszek Lisowski2, David MacDonald1, Szun Szun Tay1, Claire McGuffog1, Mehdi Ramezani-Moghadam1, Jelte Krol1, Geoffrey W. McCaughan1, David G. Bowen1, and Patrick Bertolino1

1Liver Immunology Program and AW Morrow Gastroenterology and Liver Centre, Centenary Institute, Royal Prince Alfred Hospital and University of Sydney, NSW, Australia. 2Translational Vectorology Group and Vector and Genome Engineering Facility, Childrens Medical Research Institute, NSW, Australia Background: We have previously established that factors influencing primary T cell activation, such as antigen load in the liver, will affect the final outcome of CD8 T cell responses. CD8 T cells activated under a high antigen load fail to clear liver-specific antigens and become exhausted or deleted, similarly to CD8 T cells during chronic human liver disease. Interestingly the binding strength of a T cell’s receptor to a peptide:MHC complex, an important factor influencing primary activation, has been poorly studied in this context. Aim & Objectives: To investigate how the binding affinity of the TCR for its ligand influences the activation, expansion, and fate of CD8 T cells recognising liver-expressed antigens. Methods: CD45.1+ CD8 T cells expressing a transgenic TCR (OT-1) were CTV-labelled and transferred into CD45.2+ mice. Recipients were inoculated 24 hours later with a high dose of a recombinant adeno-associated viral (rAAV) vector targeting hepatocytes and encoding for wildtype OVA or altered OVA variants (recognised with varying affinities by the donor T cells). The phenotype and proliferation of donor CD8 T cells was assessed by flow cytometry. Results: Activation and expansion of CD8 T cells was positively correlated with TCR:pMHC binding affinity. Although CD8 T cells activated by low or intermediate affinity ligands proliferated, all donor cells were deleted within 21 days. While CD8 T cells activated by high affinity ligands also underwent deletion in most recipient mice, some recipient mice retained CD8 populations that over-expressed PD-1, suggesting exhaustion. Conclusion: This study suggests that under a high antigen load, intrahepatic CD8 T cell activation favours the expansion and survival of high affinity clones, despite surviving cells becoming exhausted. This knowledge is critical to design more effective treatments of chronic liver diseases in which liver CD8 T cells are functionally exhausted.


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.6 ARC (APOPTOSIS REPRESSOR WITH CASPASE RECRUITMENT DOMAIN) DEFICIENT MICE DEVELOP SEVERE POST-TRAUMATIC KNEE OSTEOARTHRITIS BUT HAVE REDUCED PAIN Shihani P Stoner1,3,Cindy C Shu1,3, Yolanda Liu1,3, Susan M Smith1,3, Anthony W Ashton2,3, Christopher B Little1,3

1 Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research 2 Perinatal Medicine Research 3 Kolling Institute, Royal North Shore Hospital, Faculty of Medical Health, University of Sydney Background: Healthy articular chondrocytes express abundant ARC mRNA and protein, which are lost with spontaneous/age-associated and post-traumatic osteoarthritis (ptOA). In vitro ARC overexpression protects against chondrocyte apoptosis and prevents hypertrophic differentiation. Aim & Objectives: Given ARC regulates chondrocyte differentiation and survival, we investigated the effect of ARC depletion on surgically induced ptOA in mice. Methods: OA was induced by destabilisation of the medial meniscus (DMM) in wildtype (C57BL6) and ARC-deficient (Nol3-/-) mice. At 4/8/16 weeks post-DMM, operated knee joints were collected for histopathology and synovial tissue isolated for gene expression analyses. Pain sensitisation (distal mechanical allodynia and local hyperalgesia) was assessed from baseline in the 16-week group. Results: All animals developed mechanical allodynia and local hyperalgesia post-DMM. By 4 weeks Nol3-/- mice had significantly less allodynia than wildtype, and this was maintained to 16 weeks. Local hyperalgesia followed a similar trend although did not reach significance. Histopathology scoring revealed no difference between genotypes in synovitis, subchondral bone remodelling or osteophytes, but Nol3-/- mice developed increased chondrocyte hypertrophy by 8 weeks, and significantly greater cartilage structural damage at 16 weeks compared to wildtype. Both genotypes had similar initial upregulation of Mmp2/3/13, while Nol3-/- demonstrated reduced initial increase of Adamts4/5 at 4 weeks; by 16 weeks Mmp2 and Adamts4 remained increased in Nol3-/-. Conclusion: Increased chondrocyte hypertrophy and MMP expression may have exacerbated late-stage cartilage erosion in Nol3-/- mice, but the mechanism driving pain reduction remains to be elucidated. Prolonging chondrocyte ARC expression may aid in delaying/reducing ptOA progression.

Abstract 1.7

ELUCIDATING THE ROLE OF OVEREXPRESSED N 6-METHYLADENOSINE METHYLTRANSFERASE, VIRMA, IN BREAST CANCER. Natalia Pinello1,2, Michelle van Geldermalsen1,2, Chau-To Kwok1,2, Justin JL Wong1,2

1 Epigenetics and RNA Biology Program, Centenary Institute, University of Sydney, Camperdown 2050, NSW, Australia. 2 Sydney Medical School, University of Sydney, Camperdown 2050, NSW, Australia. Background: N 6-methyladenosine (m6A) is the most abundant modification of mRNAs. It is deposited on mRNAs by a methyltransferase complex which consists of methyltransferase-like proteins 3 and 14, WTAP, VIRMA, RBM15 and Zc3h13. Dysregulation of m6A homeostasis has been implicated in human malignancies including breast cancer. VIRMA is overexpressed in breast cancers indicating that m6A dysregulation may play a role in breast tumorigenesis. Aim & Objectives: We sought to characterise the hitherto unknown role of abnormal VIRMA expression in breast cancer. Methods: We analysed the Human Genome Atlas Research Network (TCGA) and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) data to determine genetic aberrancies that affect VIRMA in breast cancers. We performed shRNA-mediated VIRMA knockdown in four breast cancer cell lines, MCF7, HS578T, MD453, SKBR3. We assessed cell proliferation (MTT assay) and clonogenicity after confirming VIRMA knockdown in cells. The presence and relative abundance of m6A RNA methylation was determined by Dot Blot using an anti-m6A antibody. Results: We identified amplification of the VIRMA gene in approximately 15% of breast cancers in association with poorer overall survival in both the TCGA and METABRIC cohorts (P<0.035, Log-Rank Test). Upon VIRMA knockdown, we observed a marked decrease in cell proliferation and clonogenicity in all breast cancer cell lines tested. Consistently, we observed a decrease in RNA m6A levels in VIRMA knockdown cells. Conclusion: These results indicate a potential role of VIRMA overexpression in breast tumorigenesis. Future work may provide insights into the usefulness of targeting VIRMA for cancer therapy.


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.8 DIFFERENTIAL CHEMOKINE RECEPTOR EXPRESSION AND USAGE BY IMMEDIATE DENDRITIC CELL PRECURSORS Stuart J Cook, Quintin Lee, Alex CH Wong, Benjamin C Spann, Jonathan N Vincent, Justin JL Wong, Andreas Schlitzer, Mark D Gorrell, Wolfgang Weninger, Mainthan Palendira and Ben Roediger Skin Imaging and Inflammation Laboratory, Centenary Institute, Royal Prince Alfred Hospital and University of Sydney, NSW, Australia Conventional dendritic cells (cDCs) derive from distinct precursor cells called pre-DCs in the bone marrow, which traffic through the blood to the periphery. Coordinated trafficking of these cells is necessary for different immune challenges. For instance, Type 1 cDCs (cDC1) are important for immune responses to viral infection and cancer due to their unique ability to cross present antigen and prime T helper type 1 (Th1) immunity. We have recently shown their precursor, pre-cDC1s, express and use the chemokine receptor CXCR3 to traffic to melanoma tumours in mice and their kinetics can be altered using inhibitors of dipeptidyl peptidase-4 (DPP4). However, whether pre-cDC1s express and use CXCR3 to traffic to tumours in humans has not been demonstrated. Here, I will present new data showing HLA-DR+ CD45RA+ CD123– CADM1hi CD1c– pre-cDC1 express CXCR3 in human blood distinctly from HLA-DR+ CD45RA+ CD123– CADM1lo CD1c+ pre-cDC2. CXCR3 expression was maintained in mature CD45RA– CD45RO+ CADM1hi CD1c– cDC1 in both healthy human blood and in melanoma. Collectively, our results suggest a selective role for CXCR3 within the cDC1 lineage in both mouse and man, which holds relevance for the development of novel cancer immunotherapies.

Abstract 1.9 MODULATION OF CATION INTERACTIONS WITH ZWITTERIONIC PHOSPHOLIPID BILAYERS BY HEADGROUP ASSOCIATED CARBONYL OXYGENS. Alvaro Garcia1, Beatriu Domingo Tafalla1, Bruce Cornell2, Charles G. Cranfield1

1 School of Life Sciences, University of Technology Sydney, NSW, Australia. 2 SDx Tethered Membranes Pty. Ltd., Unit 6, 30-32 Barcoo Street, Roseville, NSW 2069, Australia. Background: Despite the many reported studies of mono and divalent cation interactions with phospholipid bilayers, the structure of the phospholipid-water interface and its modulation by ions remains poorly understood. Determining the phospholipid headgroup constituent involved in these interactions would enhance our understanding of this important phenomenon. Aim & Objectives: The aim of the present study is to examine the role of the phospholipid ester moiety in modulating the interaction of sodium, potassium, calcium and magnesium with zwitterionic phospholipid bilayers. Methods: We used DOPC (ester linked) and diether-DOPC (ether linked) phospholipids to determine the effect of headgroup associated carbonyl oxygens on cation interactions with bilayers. We used a monolayer coating chemistry comprised of benzyl disulphide eleven-oxygen-ethylene-glycol reservoir linkers with a C20 phytanyl group as tethers on gold to anchor the tethered bilayer. Coupled with electrical impedance spectroscopy, this enabled us to determine changes in the electrical properties of tethered membranes. Results: We report an ester dependence on the association of Na+ and Ca2+ to the phospholipid headgroup. This ester dependence is absent in the interactions observed with K+ and Mg2+. Ca2+ demonstrates a higher affinity for the phospholipid bilayer than Mg2+ and modulates the overall membrane conduction at concentrations that are physiologically relevant. Conclusion: The phospholipid headgroup associated carbonyl oxygens play an integral role in modulating cation interactions with phospholipid bilayers. The fact that Ca2+ has the ability to modulate bilayer properties at physiologically relevant concentrations suggests that Ca2+ fluxes may indirectly modulate the function of membrane-associated proteins in cells.

ABSTRACT SESSION 2: EARLY CAREER RESEARCHERS’ TALKS I


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.10 TISSUE-SPECIFIC ROLES OF SIRT1 IN THE DEVELOPMENTAL ORIGIN OF METABOLIC DISORDERS DUE TO MATERNAL HIGH-FAT FEEDING Long T. Nguyen1,2*, Crystal Mak2, Hui Chen2, Amgad Zaky1, Carol Pollock1 and Sonia Saad1

1 Renal medicine, Kolling Institute, Royal North Shore Hospital, University of Sydney, Sydney, New South Wales, Australia 2 School of Life Sciences, Faculty of Science, University of Technology Sydney, Sydney, New South Wales, Australia Background: Obesity in pregnancy is an emerging risk factor of metabolic disorders in the offspring. Sirtuin (SIRT)1, a potent regulator of lifespan, metabolism and stress responses, has been shown to be implicated in this programming effect. Aim & Objectives: We aimed to study how SIRT1 is regulated in different organs in the offspring in association with maternal obesity-induced metabolic disorders. We also examine the protecting effects of SIRT1 overexpression in early ages. Methods: Female mice were fed a control or high-fat diet (HFD) prior to and throughout gestation and lactation. SIRT1-overexpressed offspring were produced by mating the mice with hemizygous SIRT1-transgenic male breeders. Hypothalamus, adipose tissues, liver and kidney were collected for assessment of metabolic signalling and stress responses. Results: Maternal HFD led to reduced expression of SIRT1 in hypothalamus, liver and kidney. SIRT1 overexpression in offspring from obese dams reversed their body weight, adiposity and insulin resistance, and improved lipid metabolism and redox balance in liver and kidney. However, appetite and urinary albumin/creatinine ratio were not affected. Conclusion: Our study provides causal, tissue-specific roles of SIRT1 in the developmental origin of metabolic disorders due to high-fat diet in pregnancy and illuminates SIRT1 as a promising clinical target to prevent obesity and related disorders in offspring of obese/overnourished mothers. Abstract 1.11 INHIBITION OF THROMBOCYTE ACTIVATION RESTORES PROTECTIVE IMMUNITY TO MYCOBACTERIAL INFECTION Elinor Hortle1, Khelsey Johnson2, Jordan Shavit3, David Tobin2, Stefan Oehlers*1,4

1 Tuberculosis Research Program, Centenary Institute, Sydney, Australia 2 Department of Molecular Genetics and Microbiology, Center for Microbial Pathogenesis, Duke University School of Medicine, Durham, North Carolina, USA 3 Department of Pediatrics and Communicable Diseases, University of Michigan, Ann Arbor, Michigan, USA 4 Sydney Medical School, University of Sydney, Sydney, Australia * Corresponding author Background: Infection with M. tuberculosis (TB) causes a hyper-coagulable state, along with an increase in the number of circulating platelets. In vitro evidence suggests that these platelets may support bacterial survival within the host by converting macrophages into foam cells; we therefore hypothesise that anti-platelet drugs may be an effective host-directed therapy to aid in the treatment of TB. Aim & Objectives: To determine whether anti-platelet drugs can reduce mycobacterial burden, and if so, determine if this effect is macrophage dependent. Methods: Here we used the zebrafish/ M. marinum model of TB infection. This model resembles TB granuloma formation more closely than mouse models. Zebrafish larvae are also transparent, so fluorescently tagged bacteria and cells can be used to observe platelet involvement in infection in real time in a live animal. Fish were infected with M. marinum, and treated with anti-platelet drugs for a period of 5 days. Results: We found that infection-induced coagulation and thrombocytosis are conserved in the zebrafish-M. marinum infection model. The anti-platelet drugs – aspirin, tirofiban, and eptifibatide - were able to reduce bacterial burden by 50%, and this reduction was both platelet and macrophage dependent. Drug treatment was associated with reduced lipid accumulation and cell death at sites of bacterial growth, suggesting less conversion of macrophages into foam cells, and less necrosis within the granuloma. Conclusion Our results indicate that thrombocytes have a pathogenic role in mycobacterial infection, and that anti-platelet drugs may be effective host-directed therapies to aid in the treatment of TB.


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MONDAY 19TH NOVEMBER 2018 (continued) Abstract 1.12 IMPLEMENTATION OF THE GOAL-DIRECTED MEDICATION REVIEW ELECTRONIC DECISION SUPPORT SYSTEM (GMEDSS) INTO HOME MEDICINES REVIEW (HMR) TO DEPRESCRIBE MEDICATIONS IN OLDER ADULTS Lisa Kouladjian O’Donnell1, Mouna Sawan1, Emily Reeve1, Danijela Gnjidic2, Timothy Chen2, Patrick Kelly3, J Simon Bell4, Sarah Hilmer1

1 NHMRC Cognitive Decline Partnership Centre, Kolling Institute, Sydney University, Royal North Shore Hospital St Leonards, NSW 2 Sydney University Faculty of Medicine and Health, School of Pharmacy, Camperdown, NSW 3Sydney University Faculty of Medicine and Health, School of Public Health, Camperdown, NSW 4Faculty of Pharmacy and Pharmaceutical Sciences, Monash University, Melbourne, VIC Introduction: People with dementia living in the community are prescribed more medicines compared to people without dementia, and may be vulnerable to adverse effects (e.g. confusion) from high-risk medicines (e.g. anticholinergics and sedatives, as measured using Drug Burden Index (DBI)). Computerised Clinical Decision Support Systems (CCDSS) have demonstrated effectiveness in improving appropriate prescribing in older adults. HMR is a government-funded pharmacist-led medicines review service that aims to increase patient benefits from medicines. Aim: To describe the preliminary baseline data on participants characteristics and DBI from a cluster-RCT of a CCDSS in HMRs to reduce the proportion of patients (with and without dementia) who are exposed to DBI medicines. Methods: This study is a two-arm, parallel group, cluster-RCT, clustered at the level of accredited pharmacists (AP). The GMEDSS is a CCDSS that incorporates validated deprescribing tools and guides (e.g. DBI, Patient Attitudes Towards Deprescribing questionnaires, and Goals of Care) applicable to patients with and without dementia and their carers. APs were randomised into the intervention (HMR + GMEDSS) or control (HMR only) groups. APs collected data (e.g. medication profile) from up to 10 of their HMR patients at baseline (during HMR interview) and at 3-months follow-up. Results: To date, 53 APs have been enrolled in the study, recruiting 179 patients. Preliminary analysis of baseline data (n=161): Mean age (±SD) of all patients was 77.7±7.4 years and 86% of patients were female. Proportion of patients with DBI>0 was 64.2% and 80.0% in the control and intervention groups, respectively. Median DBI (IQR) for the control group (n=81) is 0.5(0-1) and intervention group (n=80) is 0.78 (0-0.90). Conclusion and Relevance: This preliminary baseline data will form part of an analysis of whether addition of the GMEDSS intervention into HMR will reduce anticholinergic and sedative medication exposure.


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TUESDAY 20TH NOVEMBER 2018

Abstract 2.1 INTRAUTERINE CIGARETTE SMOKE EXPOSURE INCREASES METABOLIC AND LIVER DAMAGE WHICH WERE REVERSED BY MITOCHONDRIAL ANTIOXIDANT Gerard Li1, Brian Oliver1,2, Sonia Saad1,3, Hui Chen1,

1 School of Life Sciences, Faculty of Science, University of Technology Sydney, NSW, Australia 2 Respiratory Cellular and Molecular Biology, Woolcock Institute of Medical Research, Sydney, NSW, 2037, Australia 3 Kolling Institute of Medical Research, Royal North Shore Hospital, University of Sydney, St. Leonards, New South Wales, Australia Background: Intrauterine cigarette smoke exposure increases the risk of metabolic disorders in the offspring, which could be due to mitochondrial oxidative stress damage. MitoQ is a potent mitochondrial targeted antioxidant which could minimise these damages. Aim & Objectives: We aimed to determine if maternal MitoQ supplementation during pregnancy could ameliorate metabolic disorders in the male offspring due to in utero cigarette smoke exposure. Methods: Female Balb/C mice were divided into three groups: Sham (exposed to air), SE (exposed to cigarette smoke before mating, during gestation and lactation), SEMQ (SE dams with MitoQ supplementation during gestation and lactation). Intraperitoneal glucose tolerance test (IPGTT) was performed in the male offspring at 12 weeks of age. Livers and plasma were collected at 13 weeks of age. Data were analysed with one-way ANOVA with Fisher’s Least Significant Difference post hoc test. Results: SE offspring exhibited reduced glucose tolerance and increased hepatic triglyceride accumulation. Furthermore, there was an increase in plasma ALT activity and increased inflammation indicating liver damage. This was associated with increased hepatic mitochondrial ROS and reduced mitochondrial density. Additional MitoQ supplementation in the pregnant dams reduced mitochondrial ROS level and increased endogenous mitochondrial antioxidants and markers of mitophagy and mitochondrial biogenesis. Overall, this restoration of mitochondrial integrity markers normalised liver triglyceride concentration, systemic glucose metabolism and markers of liver damage. Conclusion: MitoQ supplementation reversed metabolic and hepatic damage in the offspring due to maternal cigarette smoke exposure and could be an option among pregnant women who are unable to quit smoking. Abstract 2.2 EFFECT OF CHRONIC POLYPHARMACY AND THE DRUG BURDEN INDEX (DBI) ON MUSCLE FUNCTION AND STRUCTURE IN AGED MICE. Gizem Gemikonakli1,2, John Mach1,2, Trang Tran1,2, Susan Howlett3, Rafael de Cabo4, David G Le Couteur2,5& Sarah N Hilmer1,2 1 Lab of Ageing and Pharmacology, Kolling Institute, Royal North Shore Hosp, Sydney, NSW, Australia. 2 Northern Clinical School, Univ of Sydney, NSW, Australia. 3 Dalhousie University, Halifax, Canada. 4 Translational Gerontology Branch, National Institute on Aging, Maryland, USA. 5 ANZAC Research Institute, Sydney, NSW, Australia. Background: Ageing, polypharmacy (use of ≥5 medications) and increasing DBI (total anticholinergic/sedative medication exposure) are associated with falls and impaired physical function. Preclinical ageing models can assess underlying mechanistic changes. Aim & objectives: We investigated whether chronic polypharmacy or monotherapy, with increasing DBI and/or cessation (deprescribing), affected physical function and/or muscle histology in mice. Methods: 12-month-old male C57BL/6 mice received either control diet or study drug(s) at therapeutic doses. Polypharmacy diets consisted of zero-DBI (metoprolol, simvastatin, omeprazole, paracetamol, irbesartan), low-DBI (metoprolol, simvastatin, omeprazole, paracetamol, citalopram) and high-DBI (metoprolol, simvastatin, citalopram, oxycodone, oxybutynin). Individual drugs (high-DBI regimen) were tested as monotherapy. At 21-months, animals were randomised to continue treatment or gradual withdrawal. Rotarod performance was assessed at 12-24-months, and balance beam at 24-months. Gastrocnemius muscle samples were collected at 26-months. Results: Rotarod performance at 21-24-months indicated increased endurance for metoprolol mice (n=15-36) compared to control (n=24-29) and high-DBI (n=18-38), and for zero-DBI (n=15-34) compared to low-DBI (n=19-40); (p<0.05). Balance assessment showed deprescribed high-DBI (n=18) and metoprolol (n=16) mice performed better than their prescribed comparators (n=10-15; p<0.05). Preliminary histology results suggest a trend towards less muscle fibres per field in control (n=3), compared to high-DBI (n=3; p=0.119) and citalopram (n=2; p=0.049), while collagen quantification suggests no difference between control (n=5), high-DBI (n=6) and high-DBI deprescribed (n=4). Conclusion: Rotarod detected differences between drug treatments and deprescribing affected balance. Our preclinical results suggest certain polypharmacy and monotherapy regimens impact measures of muscle function and may affect structure. Future research will complete muscle histological assessment.

ABSTRACT SESSION 3: HIGHER DEGREE RESEARCH STUDENTS’ TALKS I


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.3 MACROPHAGE DEVELOPMENT AND ACTIVATION INVOLVES COORDINATED INTRON RETENTION IN KEY INFLAMMATORY REGULATORS Immanuel D Green1,2, Natalia Pinello1,2, Renhua Song1,2, John EJ Rasko2,3,4, Justin J-L Wong1,2 1 Epigenetics and RNA Biology Program, Centenary Institute, University of Sydney, Camperdown 2050, NSW, Australia 2 Sydney Medical School, University of Sydney, Camperdown 2050, NSW, Australia 3 Gene and Stem Cell Therapy Program, Centenary Institute, University of Sydney, Camperdown 2050, NSW, Australia 4 Cell and Molecular Therapies, Royal Prince Alfred Hospital, Camperdown 2050, NSW, Australia Background: Intron retention (IR) is a form of alternative splicing that includes a non-coding intron in mature mRNA. We have previously demonstrated that IR regulates gene expression during granulocyte differentiation via cytoplasmic nonsense-mediated decay. Aim & Objectives: We investigated the roles of differential IR during macrophage development and activation. Methods: We performed mRNA sequencing and bioinformatic analysis (IRFinder) on THP-1 monocytes and THP-1-derived M0 and M1 macrophages, to measure differential IR. Gene Ontology (GO) analysis was used to determine the functional enrichment of intron-retaining transcripts. Differentially-expressed retained introns were validated using RT-qPCR. Nuclear/cytoplasmic fractionation of THP-1 monocytes and macrophages was used to establish the localisation of intron-retaining transcripts. We polarised primary M1 macrophages to validate differential IR in key genes of interest. Results: IRFinder analysis identified 881/1126 (78.2%) introns decreased in retention during monocyte-to-macrophage differentiation. Differential IR was inversely correlated with gene expression. GO analysis of transcripts with decreased IR were significantly enriched in the type I interferon-mediated signalling category (p < 0.005). Five IR events, belonging to genes involved in myeloid differentiation, adhesion, phagocytosis and chemotaxis were confirmed by RT-qPCR. A 10- to 150-fold enrichment of intron-retaining transcripts was observed in the nucleus, indicating nuclear detention may regulate gene expression. During M1 macrophage polarisation, we found a dramatic decrease in IR alongside upregulation of vital proinflammatory regulators CXCL2 and NFKBIZ. Conclusion: IR predominantly decreases in monocyte-to-macrophage differentiation. Our findings substantiate nuclear detention of intron-retaining transcripts as an important mechanism of gene expression control during the innate immune response. Abstract 2.4 RISK OF ACL RUPTURE AND OSTEOARTHRITIS: A ROLE FOR PRIOR SUB-CRITICAL INJURY Carina Blaker1,2, Sanaa Zaki2, Christopher Little2, Elizabeth Clarke1

1 Murray Maxwell Biomechanics Laboratory, Institute of Bone and Joint Research, Kolling Institute, Northern Sydney Local Health District, Faculty of Medicine and Health, University of Sydney, St. Leonards, NSW, Australia 2 Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research, Kolling Institute, Northern Sydney Local Health District, Faculty of Medicine and Health, University of Sydney, St. Leonards, NSW, Australia Background: The incidence of anterior-cruciate-ligament (ACL) rupture is increasing at a rapid rate and has been associated with considerable long-term disability through early osteoarthritis development. The reasons why only 50% of ACL injuries progress to osteoarthritis is unclear but prior mild (sub-critical) knee injuries may play an important role. Aim & Objectives: Investigate the role of sub-critical knee injury in ACL rupture risk and susceptibility to osteoarthritis. Methods: A single, sub-critical knee injury was induced in 10-week-old male C57BL/6 mice. Mice were evaluated for changes in pain responses, joint mechanics, rupture risk (ACL strength) and structural pathology associated with osteoarthritis at 1-, 2-, 4- and 8-weeks post-injury. Results: Although sub-critical knee injury did not cause unloading of the injured limb during weight-bearing there was evidence of sensitisation of the affected limb 1-8 weeks post-injury (P<0.0001). Mechanically, sub-critical knee injury had no effect on joint mechanics but was found to significantly impact ACL strength (P=0.0148). At 4- and 8-weeks post-injury ACL strength was reduced by 14-16% compared to un-injured controls. Sub-critical injury also induced focal lesions in both the cartilage and subchondral bone within the patellofemoral and tibiofemoral knee compartments. Conclusion: A sub-critical knee injury did not cause adverse pain or altered mechanics but did lead to sensitisation, an increased risk of rupture and joint lesions which may be susceptible to osteoarthritic progression. These results highlight the importance of addressing sub-critical injuries as risk factors for both ACL injury and osteoarthritis, and may improve identification of at-risk patients for early intervention.


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2. 5 THE ROLE OF EXTRACELLULAR VESICLES IN THE MODULATION OF ENDOTHELIAL JUNCTIONS IN AN IN VITRO MODEL OF CEREBRAL MALARIA Benjamin Sealy1, Iris Cheng1, Valery Combes1

1 School of Life Sciences, Microvesicles and Malaria Research Group, University of Technology Sydney, NSW, Australia Aims: Extracellular Vesicles (EVs) are functional sub-cellular fragments capable of transferring surface markers, proteins, and nucleic acids to cells that internalise them. Recent studies have suggested that EVs participate in the development of the vascular lesion during Cerebral Malaria (CM). Using an in vitro Blood-Brain barrier (BBB) model, we aim to investigate the effects EVs have on the modulation of endothelial integrity by measuring the expression of junctional protein, VE-cadherin and the activation status of the endothelial monolayer. EVs released by both normal and Plasmodium falciparum-parasitised red blood cells (nRBCs, pRBCs) after 48h culture were purified from the supernatant by sequential centrifugation. Methods: Primary human brain endothelial cells (HBECs) were incubated with nRBCs, pRBCs, nRBC-/pRBCs-EVs (nEVs, pEVs), or a combination of them. Modulation in VE-cadherin expression both local and in relation to the presence of PRBCs and/or EVs was assessed by a combination of high-content, high-resolution and OMX super-resolution microscopy. Expression of cell adhesion molecules (CAMs) was measured by flow cytometry. Results: Quantitative image analysis showed that non-parasitised conditions triggered up-regulation of VE-cadherin expression, whereas parasitised conditions resulted in a significant down-regulation. We also observed that p-EVs were taken up by HBEC at twice the rate of n-EVs. Expression of CAMs was increased in the presence of p-MVs and PRBC-Mix. Conclusion: These results suggest that interactions between EVs and their cells of origin have similar effect of target cells. Further studies are currently ongoing in order to determine which molecular pathways are involved in the changes observed. Abstract 2.6 CHRONIC ALCOHOL DRINKING ALTERES MICRORNA IN SERUM IN PATIENTS WITH LIVER CIRRHOSIS Fathima Shihana1, Devanshi Seth2,3,4 1Discipline of Pharmacology, Sydney Medical School, The University of Sydney, NSW, Australia. 2 Drug Health Services, Royal Prince Alfred Hospital, Camperdown, NSW, Australia 3 Discipline of Clinical Medicine & Addiction Medicine, Faculty of Medicine, The University of Sydney, NSW, Australia 4The Centenary Institute of Cancer Medicine & Cell Biology, The University of Sydney, NSW, Australia Background: Alcoholic Liver Cirrhosis (ALC) is the major medical complication of prolonged drinking associated with underlying addiction issues. Recent literature suggests that microRNAs may be important players in alcohol use disorders, including ALC. Few microRNAs associated with chronic alcohol use and ALC are known, but their action is not clear in human ALC. Aim & objective: To evaluate global serum microRNAs as risk factors for ALC in heavy drinkers. Methods: Serum samples from age and gender-matched chronic alcohol drinkers (without liver disease, Controls=12; with liver cirrhosis, Cases=12) were used for microRNA profiling (TaqMan OpenArray ultra-high content platform). Identified miRNAs were validated in an independent set of patients (Controls=12, Cases =12), matched for age and gender. Ingenuity was used for target gene and network identification. Results: Novel microRNA signature was identified in drinkers distinguishing patients with liver cirrhosis from those without liver disease. Fourteen microRNAs were significantly downregulated in Cases compared to Controls in the discovery cohort (p<0.05, Ct >1.5). Validation in independent cohort confirmed downregulation of 7 common microRNAs (miR-27b,1, miR-29c, miR-130a, miR-191, miR-221, miR-484 and miR-335) and identification of another 7 miRNAs, significantly downregulated (p<0.05, Ct >1.5) in Cases vs Controls. Majority of microRNAs were specific to female Cases in both discovery and replication cohort. Conclusion: Serum microRNAs can potentially be used to identify drinkers at risk of cirrhosis. Since females are known to be at a greater risk of developing liver cirrhosis, it is intriguing to find majority of dysregulated microRNAs significantly different only in female patients.


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TUESDAY 20TH NOVEMBER 2018 (continued)

Abstract 2.8 MIR-99B AND MIR-652 ARE REQUIRED FOR OPTIMAL CONTROL OF M. TUBERCULOSIS INFECTION IN HUMAN MACROPHAGES Jessica L Pedersen1, Simone Barry2, YuRong Yang3, Xiaolin Wang4, Warwick Britton2, Nilesh J Bokil1 and Bernadette Saunders1

1 School of Life Science, University of Technology Sydney, NSW, Australia 2 Centenary Institute, The University of Sydney, NSW 2042 Australia 3 Ningxia Medical University, Yinchuan, Ningxia, P.R. China 4 Infectious Disease Hospital of Ningxia, Yinchuan, P.R. of China Background: New tools including diagnostics and therapeutics are urgently required to decrease the global tuberculosis (TB) burden, with over 10 million new cases in 2017 and 1.6 million deaths. Analysis of miRNA expression in the plasma of 100 active TB patients and healthy controls, identified 87 microRNA that were differentially regulated. A subset of 5 of these microRNA distinguished the two populations with a sensitivity of 94% and specificity of 88%. Aim & Objectives: This study investigated the role of two of these microRNA, miR-99b and miR-652 in human macrophages. Methods: Expression of miR-99b and miR-652 in human macrophages during M. tuberculosis infection was assessed by qPCR. The levels of these microRNA were then modulated by introducing antagomirs, and the impact on macrophage function during infection was examined. Results: miR-99b was significantly up-regulated in the plasma of TB patients compared to healthy controls at the time of diagnosis, whereas miR-652 was significantly down-regulated. Similarly, in human macrophages infected with M. tuberculosis miR-99b expression was significantly up-regulated while miR-652 expression remained significantly down-regulated. Interestingly however, inhibition of either miR-99b or miR-652 in macrophages led to a significant increase in bacterial burden after 24 hours infection. Evaluation of the precise mechanisms of action of miR-99b and miR-652 in regulating host response to M. tuberculosis infection is underway. Conclusion: This study suggests that both miR-99b and miR-652 are required for optimal control of TB infection and may potentially be new therapeutic targets for anti-TB therapy. Abstract 2.9 PRPF19 REGULATES INTRON RETENTION IN ACUTE MYELOID LEUKAEMIA Alex CH Wong1,2,3, John EJ Rasko1,2,4, Justin J-L Wong1,2,3 1 Gene & Stem Cell Therapy Program, Centenary Institute, University of Sydney, Camperdown 2050, Australia 2 Sydney Medical School, University of Sydney, NSW 2006, Australia 3 Epigenetics and RNA Biology Program, Centenary Institute, University of Sydney, Camperdown 2050, Australia 4 Cell and Molecular Therapies, Royal Prince Alfred Hospital, Camperdown 2050, Australia Background: Pre-mRNA-processing factor 19 (PRPF19) catalyses conformational changes in the spliceosome. PRPF19 is also essential for recognition, processing and repair of DNA double-stranded breaks. PRPF19 confers chemoresistance in many cancers and reducing PRPF19 expression inhibits cell proliferation and clonogenicity. The role of PRPF19 in regulating intron retention (IR) in cancer remain uncharacterised. Aim & Objectives: We examined whether PRPF19 regulates IR using shRNA knockdown of PRPF19 in the THP-1 myeloid leukaemia cell line. Methods: THP-1 cells were transduced with lentiviral tetracycline-inducible shRNA targeting PRPF19. Doxycycline-treated cells were harvested at 24 & 48 hours, compared to vehicle control. mRNAseq was analysed and IR measured using IRFinder. Differential IR events were identified using DESeq2, with Benjamini-Hochberg correction for multiple testing. Gene ontology analysis was performed using Panther. Results: PRPF19 knockdown induced dramatic increases in IR, with 2584 introns showing increased IR at 48 hours following PRPF19 knockdown (adjusted p-value < 0.05), compared with 73 introns with decreased IR. Pathways affected by increased IR include RNA processing/splicing and DNA replication/repair. Interestingly, ultra-short introns were predominantly retained, with 976 of the 2584 introns with increased IR being shorter than 100 base pairs. The DNA damage repair gene RECQL4 contains multiple ultra-short introns which were retained following PRPF19 knockdown. This is the subject of ongoing studies. Conclusion: PRPF19 is not essential to splicing in humans but is an obligate splicing factor for a specific subset of ultra-short introns. The coupling of DNA damage response to mRNA splicing in cancer may be mediated by PRPF19-regulated IR.

ABSTRACT SESSION 4: HIGHER DEGREE RESEARCH STUDENTS’ TALKS II


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.10 BRINGING BACK THE OLD – OUABAIN A PROMISING TREATMENT FOR NEW-ONSET ATRIAL FIBRILLATION? Yeon Jae Kim1, Elisha Hamilton1, William Hannam1, Chia-Chi Liu1, Helge H Rasmussen1, 2, Alvaro Garcia3

1 Kolling Institute, University of Sydney, NSW, Australia 2 Department of Cardiology, Royal North Shore Hospital, NSW, Australia 3 School of Life Sciences, University of Technology, NSW, Australia Background: Pharmacological treatments of atrial fibrillation (AF) must address pathologically elevated cardiomyocyte cytosolic Ca2+ concentrations ([Cai]). [Cai] is indirectly maintained by the Na+-K+ pump (NKP). While cardiotonic steroids (CTSs) inhibit the NKP, sporadic reports of low-dose ouabain-induced stimulation exist. Such stimulation should reduce [Cai]. Aim & Objectives: To observe whether CTS-induced NKP stimulation occurs, and identify the mechanism and features of the CTS’s steroid core, sugar moiety, and lactone ring required for stimulation. Methods: Rabbit cardiomyocytes were extracted, voltage clamped, then exposed to experimental solutions, which blocked non-NKP currents. NKP current (Ip) was measured, with or without prior exposure to CTSs at different low concentrations. One-way ANOVAs were then performed. Results: Exposure to 5nM ouabain maximally increased Ip to 0.69±0.09 pA/pF (N=6) vs. 0.46±0.03 pA/pF in 25 controls (p<0.01). Ouabain antagonist rostafuroxin abolished this stimulation (Ip=0.40±0.04 pA/pF, N=7). NKP activity is regulated by glutathionylation, a reversible post-translational modification; cysteine-lacking FXYD3 inclusion in patch solutions, which prevents de-glutathionylation, abolished ouabain-induced stimulation. 1nM dihydroouabain (ouabain with a saturated lactone) increased Ip to 0.65±0.02 pA/pF, N=6 (p<0.01). 10-500nM ouabagenin (ouabain without a sugar moiety) did not increase Ip, nor did 5–30nM digoxin (which only shares ouabain’s lactone). 1h ouabain pre-exposure to achieve steady-state binding reduced Ip (p<0.05) at 10nM. Conclusion: Some CTSs induce transient NKP stimulation. A sugar moiety is critical for stimulation, which occurs through de-glutathionylation, a pathway previously not implicated in CTSs’ effects on NKP activity. Ouabain-induced NKP stimulation outlined may manage new-onset AF. A human trial should be considered. Abstract 2.11 WE ARE WHAT WE EAT: IDENTIFYING A REGULATORY CROSSTALK BETWEEN CENTRAL CARBON METABOLISM AND CELL DIVISION IN BACTERIA Riti Mann1, Amy L Bottomley1, Leigh G Monahan1, Elizabeth J Harry1 1 The ithree Institute, University of Technology Sydney, Ultimo, Australia Background: Cell division, an essential process in bacteria, is driven by a cytoskeletal ring structure - the Z ring, whose formation at midcell site must be tightly regulated to ensure faithful cell division. Several mechanisms have previously been described that influence the positioning and timing of Z ring assembly, but one important yet poorly understood aspect of cell division regulation is the need to coordinate division with nutrient availability. Aim: To understand the role of central carbon metabolism (CCM) in Z ring positioning in the bacterium Bacillus subtilis Methods: To investigate Z ring positioning, fluorescence microscopy was performed. The rate of DNA replication was measured using qPCR. Results: A mutant of the glycolytic enzyme pyruvate kinase (pyk) was shown to form acentral Z rings. This Z ring placement defect, however, was rescued when pyruvate was added to the growth medium, signifying pyruvate to be a specific key metabolite in coordinating cell growth with division by regulating midcell Z ring formation. We have shown that this mutant has a DNA replication defect, which is rescued after pyruvate addition. This observation raises the possibility that pyruvate is involved in cell division regulation via its effect on the DNA replication process. Conclusion: We hypothesize that the metabolic regulation of the division process is indeed an outcome of the effect that metabolic perturbation has on the DNA replication process. Understanding this interconnection between various cell cycle events not only answers as to how CCM and division are linked, but also uncovers a new area of interest for possible exploitation for the development of antimicrobial agents.


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.12 CD4 T CELL HELP IS REQUIRED FOR ROBUST CD8 T CELL RESPONSES TO HEPATOCYTE EXPRESSED ANTIGEN Kieran English1, Jelte Krol1, Yik Chun (Michael) Wong1, Szun Szun Tay1,2, Nicole A. W. Wood1, Claire McGuffog1, Ian E. Alexander2, Leszek Lisowski3, Geoffrey W. McCaughan1, David G. Bowen1,4, and Patrick Bertolino1 1 Liver Immunology Program and AW Morrow Gastroenterology and Liver Centre, Centenary Institute, Royal Prince Alfred Hospital and University of Sydney, NSW, Australia 2 Gene Therapy Research Unit, Children’s Medical Research Institute and Children’s Hospital at Westmead, NSW, Australia 3 Vector and Genome Engineering Facility, Children’s Medical Research Institute, Westmead, NSW, Australia. 4 Collaborative Transplantation Research Group, Bosch Institute, Royal Prince Alfred Hospital and University of Sydney, NSW, Australia Background: Studies show that both CD8 and CD4 T cells play critical roles in protecting humans and chimpanzees against hepatotropic pathogens, such as Hepatitis C virus, which represent major health burdens globally. While a robust anti-viral CD8+ CTL response is a predictor of resolution of hepatotropic viral infections, CD4+ helper T cells likely play an essential role in pathogen clearance by supporting CD8 T cell expansion and differentiation, a process known as "CD4 help". Aim & Objectives: Although studies have reported the determinant role of CD4 T cells in hepatotropic viral responses, the site where the help is provided and the exact mechanisms regulating CD4 help have never been elucidated. Methods: We have generated recombinant adeno-associated viral (rAAV) vectors that mediated hepatocyte specific expression of a model antigen containing both CD8 and CD4 epitopes recognised specifically by T cell receptor (TCR) transgenic CD8 T cells and CD4 T cells. This, along with antibody mediated CD4 T cell depletion, allows us to investigate how CD4 T cells influence the CD8 T cell response in the mouse liver. Results: Following rAAV treatment, CD4 T-cells are primarily activated and expanded in the liver-draining lymph nodes before accumulating in the liver. Furthermore, we show that CD4 T cells responding to hepatocyte expressed antigen are crucial for the optimal expansion of effector CTLs and the formation of a stable population of memory CD8+ T cells in the liver. Conclusion: These results suggest a model in which the CD4 T cell response boosts the number of effector virus-specific CD8 T cells during the primary immune response, resulting in enhanced clearance of antigen-expressing hepatocytes and formation of memory CD8 T cell populations. Abstract 2.13

A NOVEL THERAPEUTIC STRATEGY FOR KIDNEY FIBROSIS WITH A SMALL PROTEIN I-BODY (AD-114)

Qinghua Cao, Hao Yi, Chunling Huang, Carol Pollock, Xin-Ming Chen Renal Medicine, Kolling Institute, Royal North Shore Hospital, The University of Sydney Backgroud: Fibrosis is the final common pathway of chronic kidney disease (CKD). CXCR4 plays a central role in the development of tissue fibrosis. However, the only study using the FDA approved CXCR4 antagonist was terminated due to its off-target cardiotoxicity. Recently, a fully humanized single-domain antibody-like scaffold, termed i-body AD-114, with specific high binding affinity to CXCR4 was developed by AdAlta Ltd. AD-114 has shown anti-fibrotic effects in lung, liver and eye pathology through selectively blocking CXCR4 signaling. Aim: To define the role of i-body AD-114 in renal fibrosis. Methods: The binding of AD-114 with proximal tubular cells (PTCs) was detected using immunocytochemistry. To examine whether AD-114 blocks TGFβ1-induced fibrotic responses in PTCs, PTCs were incubated with/without TGFβ1 (2ng/ml) in the absence or presence of AD-114 (3M,4M and 5M) for 48 hours. Protein levels of CXCR4, fibronectin (FN), collagen 3 (Col 3) and collagen 4 (Col 4) and the phosphorylation of AKT and P38 were quantitated by Western blotting. Secretion of MMP-2 was quantified by ELISA. Results: TGFβ1 significantly increased the expression of CXCR4, FN, COL-3, Col 4, MMP-2 and the phosphorylation of AKT and P38 compared to normal control (P<0.001, n=4), which was reversed by AD-114 compared to control i-body (P<0.001, n=4). Conclusion: AD-114 suppresses TGFβ1-induced expression of FN, COL-3, Col 4 and MMP-2 in renal PTCs via AKT and p38 pathways. Blocking CXCR4 using the AD-114 may be a potential therapeutic strategy to CKD.


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.14 THE ROLE OF PERIVASCUAR MACROPHAGES IN CONTACT HYPERSENSITIVITY Kathy On1,2, Shweta Tikoo1,2, Rohit Jain1,2, Wolfgang Weninger1,2,3

1 Centenary Institute, Newtown, NSW Australia 2 Faculty of Medicine and Health, The University of Sydney, NSW, Australia 3 Department of Dermatology, Royal Prince Alfred Hospital, Camperdown, NSW 2050, Australia Background: Contact hypersensitivity (CHS) is a type IV hypersensitivity skin inflammatory disorder. It is initiated when small molecules, called haptens, bind to proteins in the skin resulting in their detection as foreign antigens. Although, studies have focused on the adaptive immune response, the spatiotemporal behaviour of immune cells in the skin is not very well-defined. Aim & Objectives: We aim to delineate the spatiotemporal behaviour of dermal dendritic cells (dDCs), perivascular macrophages (PVM) and mast cells (MCs) during CHS. Methods: Using intravital multiphoton microscopy, we have visualised and quantified changes in the behaviour of the above-mentioned immune cell subsets following sensitisation with the hapten 2,4-dinitro-1-fluorobenzene (DNFB). Results: Intravital imaging of homeostatic ear skin identified three migratory behaviours of dDCs within the dermis. dDCs demonstrated a predominant continuous scanning behaviour as they interacted with sessile PVM. However immediately post-sensitisation, 42.07 2.12 % of dDCs were observed to form stable interactions with PVM. These stable interactions are significantly reduced once PVM phagocytose mast cell granules (MCG) from degranulating mast cells. This suggests that MCG uptake by PVM might lead to the dissociation of PVM-DC interaction potentiating dDC egress into the draining lymph nodes. Conclusion: Our findings have identified the formation of myeloid cell clusters composed of PVM, stable dDCs and MCs. The dissociation of these clusters following MC degranulation reveals a novel cellular mechanism involved in mediating the sensitisation phase. Further investigation into the molecular mechanisms regulating these cellular events will potentiate the identification of possible therapeutic targets for CHS.

Abstract 2.15 PATTERNS OF IMMEDIATE BREAST RECONSTRUCTION IN NSW AUSTRALIA: A POPULATION-BASED STUDY Yingyu Feng1, Kathy Flitcroft1,3, Marina T. van Leeuwen4, Andrew Spillane2,3, Adam Elshaug5, Sallie Pearson4 1 School of Public Health, the University of Sydney, NSW, Australia 2 Northern Clinical School, the University of Sydney, NSW, Australia 3 Breast & Surgical Oncology at The Poche Centre, NSW, Australia 4 Centre for Big Data Research in Health, UNSW, Australia 5 Menzies Centre for Health Policy, the University of Sydney, NSW, Australia Background: The rate of immediate breast reconstruction (IBR) following mastectomy for breast cancer in Australia is low and varies between regions. To date, no previous Australian studies have examined IBR rates between all hospitals within a particular region, despite hospitals being an important known contributor to variation in IBR rates in other countries. Aim & Objectives: to examine the inter-hospital variation in IBR rates in NSW. Methods: We used cross-classified random-effects logistic regression models and extracted data on 7,961 women who underwent therapeutic mastectomy procedures in New South Wales (NSW) between January 2012 and June 2015. We also derived IBR rates by patient-, residential neighbourhood- and hospital-related factors and investigated the underlying drivers for the variations in IBR utilisation. .Results: We estimated the mean IBR rate across all hospitals that performed mastectomy during the study period to be 17.1% (95% Bayesian Credible Interval (CrI) = (12.1%-23.1%)), but observed wide variation between hospitals (variance=4.337, CrI = (2.634-6.889). Older women, women born in Asian countries (OR=0.5, CrI= (0.4-0.6)), residing in lower SES neighbourhoods (OR=0.7, CrI= (0.4-0.6) for the most disadvantaged), and who underwent surgery in public hospitals (OR=0.4, CrI= (0.1-1.0)) were significantly less likely to have IBR. Conclusion: Wide inter-hospital variation raises concerns about potential inequities in access to IBR services and unmet demand in certain areas of NSW. Explaining and quantifying the underlying causes of variation in IBR rates is the first step in identifying possible policy solutions to redress the issue.

ABSTRACT SESSION 5: EARLY CAREER RESEARCHERS’ TALKS II


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.16 CROSS-TALK BETWEEN STEM CELLS AND DISEASED CELLS IN AN OSTEOARTHRITIC JOINT: IMPLICATIONS FOR TREATMENT Jiao Jiao Li1, Christopher Little1

1 Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research, Kolling Institute, Northern Sydney Local Health District, Faculty of Medicine and Health, University of Sydney, St Leonards, NSW Background: Osteoarthritis is a leading cause of disability in aging individuals. A major predisposing factor is prior joint injury, for which current treatments only relieve short-term symptoms but do not prevent further joint degeneration. Although a significant problem, the mechanisms underlying impaired healing remain poorly understood. Aim & Objectives: To test the hypotheses that (1) impaired healing following joint injury is due to the inflammatory environment created by diseased cells, and (2) this can be reversed by ‘priming’ the d iseased cells using human mesenchymal stem cells (hMSCs) and a bioactive scaffold. Methods: hMSCs were co-cultured with diseased cells isolated from human osteoarthritic joint tissues +/- bioactive scaffold, for up to 21 days in growth, osteogenic or chondrogenic medium (simulating the post-injury joint environment). Cell interactions were assessed using RT-PCR (n=4) and histology (n=2). Results: hMSCs co-cultured with diseased cells showed significant upregulation of several markers of inflammation, matrix degradation and tissue degeneration, at both early and late time points (e.g. MMP13, TLR4). They also showed significantly impaired ability to form new bone and cartilage tissues at 21 days, but interestingly, this was partially rescued by the bioactive scaffold. Co-culture with hMSCs caused the diseased cells to undergo early and rapid downregulation of inflammatory markers (e.g. MMP13, ADAMTS5). Conclusion: Joint injury can create a highly inhibitory environment that increases inflammation in stem cells, and significantly impair their regenerative ability. ‘Priming’ the injured joint using hMSCs and/or a bioactive scaffold may efficiently neutra lise the effects of inflammation and improve subsequent treatment outcomes. Abstract 2.17 THE ROLE OF LIVER CAPSULAR MACROPHAGES IN HEPATIC FIBROSIS Tan S-Y1, MacDonald K3, McCaughan G1,2, Bowen D1, Bertolino P1

1 Liver Immunology Group, Centenary Institute, NSW Australia; 2 Department of Immunology, QIMR Berghofer Medical Research Institute, Brisbane, QLD Australia. Background: The liver contains a heterogenous population of macrophages, including tissue-resident TIM-4+CX3CR1- Kupffer cells in the liver sinusoids and TIM-4-CX3CR1+ liver capsular macrophages (LCM) in the capsule that mediate immunosurveillance in these distinct niches of the liver. Hepatic macrophages have been shown to have both pro-fibrotic and pro-restorative properties in fibrosis, but the roles of the different subsets remain unclear. The gene signature of LCM resembles that of repair macrophages, including constitutive expression of matrix metalloproteinase MMP-13, suggesting they could potentially play a role in hepatic fibrogenesis and/ or its resolution. Aim & Objectives: 1. To assess whether the number, distribution and phenotype of LCM are altered during the initiation and progression of acute liver injury and hepatic fibrosis. 2. To assess the role of LCM in the spontaneous resolution of fibrosis upon removal of the injury-causing agent. Methods: Liver fibrosis was induced by intraperitoneal administration of carbon tetrachloride for 8 weeks, or treatment with thioacetamide in drinking water (300mg/L) for 12 weeks. Multiphoton microscopy was used to co-visualise second harmonic signals from collagen matrix and hepatic macrophage subsets in fluorescent reporter lines including csf1r-GFP, cd207-GFP and cx3cr1-GFP. Results: In acute injury, the number of LCM and/or LCM precursors in the liver capsule was increased. In chronic fibrosis, LCM-like cells appeared to localise with fibrotic bands in the livers of treated animals. Ongoing studies aim to elucidate the roles of LCM in both the progression and resolution of hepatic fibrosis. Conclusion: A better understanding of the roles of LCMs and other subsets of hepatic macrophages will inform the design of macrophage-targeting anti-fibrotic strategies in the clinic.


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TUESDAY 20TH NOVEMBER 2018 (continued) Abstract 2.18 WHOLE GENOME SEQUENCING AND INTRONIC SEQUENCE INTERROGATION TO SOLVE A CASE OF EARLY-ONSET FAMILIAL PARKINSON’S DISEASE Ryan Davis1,3, Kishore Kumar1,2,3, Fabienne Edema-Hildebrand2, Brian Koentjoro1, Jin-Sung Park1, Velimir Gayevskiy3, Mark Cowley3,4, Nick Blair1,2,3, Carolyn Sue1,2,3

1 Department of Neurogenetics, Kolling Institute, University of Sydney and Royal North Shore Hospital, St Leonards, NSW 2 Department of Neurology, Royal North Shore Hospital, St Leonards, NSW 3 Translational Genomics group, Kinghorn Centre for Clinical Genomics, Garvan Institute for Medical Research, Darlinghurst, NSW 4Computational Biology, Children’s Cancer Institute, UNSW, NSW Background: Parkinson’s Disease is the second most prevalent neurodegenerative disorder (after Alzheimer’s Disease) affecting some 80,000 Australians over the age of 50. ~90% of Parkinson’s Disease presents as “sporadic” cases with no identifiable genetic cause. Of the familial Parkinson’s Diseases, mutations in the Parkin gene are particularly important and can be associated with autosomal recessive early onset disease. Aim & Objectives: Identify the underlying genetic cause of early-onset Parkinson’s Disease in a family of four female siblings. Methods: Sanger sequencing was used to screen the Parkin gene for exonic sequence variants. Western blotting for Parkin from patient fibroblasts was used to assess the protein expression levels. Whole Genome Sequencing was performed for interrogation of intronic sequences, which were analysed with a novel bioinformatic tool called Introme. Parkin transcript analysis was performed on cyclohexamide-treated fibroblasts. Results: Sanger sequencing identified a single coding variant in the mother and four siblings, a deletion of exon 2 of the Parkin gene. Western blotting revealed the absence of Parkin protein expression, indicating a second allelic variant ought to be present. Whole genome sequencing analysed with Introme revealed two candidate variants in intron 7 of the Parkin gene that created new canonical splice sites. Transcript analysis identified cryptic exon inclusion resulting from one of the identified variants. Conclusion: Interrogation of intronic sequences is of benefit in cases with strong clinical phenotype and biochemical indication where a second allelic variant cannot be identified. This approach will be useful for resolving genetic cases of this type. Abstract 2.19 FKBPL, A NOVEL PLAYER IN HEART ISCHAEMIA AND FIBROSIS Lana McClements1, Benjamin Rayner2, Abdelrahim Alqudah3, David Grieve3, Tracy Robson4

1 School of Life Sciences, University of Technology Sydney, NSW, Australia 2 Heart Research Institute, University of Sydney, NSW, Australia 3 Centre for Experimental Medicine, Queen’s University Belfast, United Kingdom 4 Department of Molecular and Cellular Therapeutics, Irish Centre for Vascular Biology, Royal College of Surgeons in Ireland, Dublin, Ireland Background: FKBPL is a critical regulator of physiological and pathological angiogenesis. In an FKBPL knockdown mouse model, impaired cardiac and vascular function were previously observed. In this study, we investigated the role of FKBPL in ischaemic heart disease and cardiac fibrosis, which are the leading causes of mortality in people with cardiovascular disease. Aim & Objectives: The aims of our study were to determine whether FKBPL has a role in cardiac ischaemia and heart fibrosis using relevant in vivo and in vitro models. Methods: FKBPL mRNA expression was determined by qPCR in hearts from rat ischaemia/reperfusion (I/R) model versus sham control at 48 h. In a separate experiment, the human cardiac fibroblast cell line T0446 was exposed to hypoxia (2% O2) or TGF-β (10 ng/mL) for 24 h or 48 h before RNA lysates were collected and FKBPL mRNA expression measured. Results: Cardiac FKBPL mRNA expression was significantly downregulated in rat I/R model (n=4, p≤.05, t-test). Interestingly, following exposure of cardiac fibroblasts to hypoxia (2%) versus normoxia (n=3, in triplicates, p≤0.01, two-way ANOVA with tukey’s post-hoc) or TGF-β versus control (n=2, in triplicates, p≤0.01 for 24 h, two-way ANOVA with tukey’s post-hoc), there was a significant increase in FKBPL mRNA expression. At 48 h, the levels of FKBPL returned to normal in both hypoxia and TGF-β conditions. Conclusion: FKBPL levels appear to be regulated by cardiac ischaemia and fibrosis in a time-dependent manner therefore changes in FKBPL expression could be explored further in these conditions as a novel pathogenic mechanism.


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POSTER PRESENTATION ABSTRACTS Abstract P1.1 DISCOVERY EXPLORATION OF MICRORNA IN HUMAN PLASMA EXTRACELLULAR VESICLES, BIOMARKERS FOR UNCOMPLICATED MALARIA Iris S. Cheng1, Nutpakal Ketprasit3, Fiona Deutsch2, Benjamin Sealy1, Nham Tran2, Duangdao Palasawan3 and Valery Combes1 1 School of School of Life Sciences, Microvesicles and malaria research group, University of Technology Sydney, NSW, Australia 2 School of Biomedical Engineering & Centre for Health Technologies, Non-coding RNA Cancer Group, University of Technology Sydney, NSW, Australia 3Chulalongkorn University, Faculty of Allied Health Sciences, Bangkok, Thailand Background: Malaria is a common tropical parasitic disease that affects two hundred million people, leading to 450,000 deaths every year. During a malaria infection, the complex life cycle of the parasite requires continuous activation and repression of various genes. MicroRNAs (miRNA) are small non-coding RNAs that have been shown to control cellular development, differentiation, metabolism, and homeostasis. Extracellular vesicles (EVs) are sub-cellular fragments released by all cell types that can transfer their content, which includes nucleic acids and proteins, into target cells, thereby regulating cell-to-cell interactions. EVs derived from human plasma contain highly stable miRNAs, the level of which are known to change in many diseases, such as malaria, thus making detection of EV-derived miRNA a potential non-invasive biomarker for uncomplicated malaria. Aim & Objectives: Analyse targeted miRNAs in EVs from P. falciparum and P. vivax malaria infected Thai patients using Real Time-quantitative PCR (RT-qPCR) to identify potential biomarkers of uncomplicated malaria. Methods: miRNA were purified from EVs extracted by sequential centrifugation of 18 P.f, 18 P.v and 20 control samples. Abundance of has-miR-451a, has-miR-150-5p, has-miR-15b-5p, miR-16-5p and has-let-7a-5p was measured. Results & conclusion: Results show significant changes in the abundance of some of these miRNA by direct comparison of CT values. Currently, housekeeping miRNA are being explored to allow better comparisons. However, these preliminary results suggest that an exploration of larger numbers of small RNA using RNAseq could expand our findings and allow the discovery of new miRNAs that could be used as potential biomarkers of malaria. Abstract P1.2 TARGETING THE E3 UBIQUITIN RING FNGER LIGASE RNF20 IN OVARIAN CANCER Kristie-Ann Dickson1,2, Alex Cole3, Rory Clifton-Bligh2, Deborah Marsh1,2 1 Medical Science, School of Life Sciences, University of Technology Sydney, NSW, Australia 2 Hormones and Cancer Group, Kolling Institute, University of Sydney, NSW, Australia 3 Department of Internal Medicine, University of Michigan Comprehensive Cancer Center, Ann Arbor, MI, USA Background: E3 ubiquitin ligases transfer ubiquitin from an E2 conjugating enzyme to a lysine on the recognized substrate. The most common family of E3 ligases are RING (Really Interesting New Gene) domain proteins. We have shown that the ring finger protein RNF20 is strongly expressed in 87% (379 of 424) of high-grade serous ovarian cancer (HGSOC) (1). The RNF20-RNF40 heterodimer is the main E3 ligase complex responsible for monoubiquitination of histone H2B at lysine 120 (H2Bub1). This active histone mark leads to an open chromatin configuration favouring DNA access by transcriptional complexes and DNA repair proteins. Aim & Objectives: To determine whether targeting the E3 ubiquitin ligase RNF20 and/or its complex member RNF40, can impact of the sensitivity of ovarian cancer cells to the chemotherapeutic platin drug cisplatin. Methods: We employed HGSOC cell line models and down-regulated RNF20 and/or its complex member RNF40, with siRNA or shRNA approaches. We explored the resulting functional effects using clonogenic cell survival assays, immunoblotting and spheroid formation. Results: We have shown that down-regulation of RNF20 decreases levels of H2Bub1 (consistent with a closed chromatin configuration); impairs the ability of HGSOC cell lines to form clones after treatment with the DNA damaging agent cisplatin; and produces spheroids of smaller size, compared to cells treated with scrambled siRNA or shRNA. Conclusion: This work provides preliminary evidence that developing strategies to therapeutically down-regulate RNF20 may increase the efficacy of standard-of-care chemotherapy for women with ovarian cancer.


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Abstract P1.3 NOVEL TARGET: AURORA KINASE INHIBITION LEADS TO TELOMERASE DOWN-REGULATION IN THYROID CANCER.

Grace Lim1,2, Martyn Bullock 1,2, Roderick Clifton-Bligh 1,2,3 1Cancer Genetics, Kolling Institute of Medical Research, St Leonards, NSW, Australia 2University of Sydney, Sydney, NSW, Australia 3Department of Endocrinology, Royal North Shore Hospital, St Leonards, NSW, Australia Background: Thyroid cancer is the most common endocrine malignancy. Whilst most thyroid cancer patients have a good prognosis, there are limited treatment options for those with aggressive disease. Systemic therapies, including the tyrosine kinase inhibitor Sorafenib are confined by dose limiting side effects, and so there is a clear near to identify new therapeutic targets. Somatic activation of telomerase, via Telomerase Reverse Transcriptase promoter (TERTp) mutations, underpins poor prognosis. The mitotic regulator Aurora Kinase B (AURKB), is overexpressed in thyroid cancer and correlates with tumour aggressiveness. Aim & Objectives: We sought to determine AURKB exerts its tumorigenic effect by regulating TERT expression. Methods: Thyroid cancer cell lines were treated with AURKB inhibitor Hesperadin, and activity on thyroid cancer pathways MAPK and PI3K/AKT/mTOR determined by western blotting (WB). Effects upon TERTp activity, TERT expression and Telomerase activity were ascertained by gene reporter, qRT-PCR and qTRAP assay respectively. Results: AURKB inhibition in thyroid cancer cells led to downregulation in both WT and mutant (C228T) TERTp activity (50%, p>0.01). Correspondingly, TERT expression levels were significantly decreased up to 72 hrs post-treatment (80%, p>0.001), and a sustained reduction in telomerase activity was observed up to 120 hours post-treatment (60%, p<0.01). Phospho-WBs revealed an inhibition of pERK and pAKT (>95%, p<0.001) between 20 mins and 9 hours post-treatment, but no significant effect on mTOR target pS6 was observed at those time-points. Conclusion: AURKB is a regulator of telomerase in thyroid cancer. Thus, AURKB warrants further investigation as a potential therapeutic target in thyroid cancer. Abstract P1.4 A SYSTEMATIC REVIEW AND META-ANALYSIS OF THE PREVALENCE OF LEFT VENTRICULAR NON-COMPACTION IN ADULTS Samantha Barratt Ross1,2, Katherine Jones1, Bianca Blanch1, Rajesh Puranik2,3, Kevin McGeechan4,5, Alexandra Barratt4,5, Christopher Semsarian1,2,3,5 1 Agnes Ginges Centre for Molecular Cardiology, Centenary Institute, Sydney, NSW, Australia 2 Sydney Medical School, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW Australia 3 Department of Cardiology, Royal Prince Alfred Hospital, Sydney, NSW, Australia 4 Sydney School of Public Health, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW, Australia 5Wiser Healthcare, Sydney, NSW, Australia Background: Assessment of heart health is common and increasingly patients undergo cardiac assessment; asymptomatic individuals may be referred for risk assessment which includes the use of new imaging modalities. Left ventricular non-compaction (LVNC) is classified as an inherited cardiomyopathy characterised by prominent trabeculations, however clinical outcomes are heterogenous and the significance of the finding in adults is unclear. Aim & Objectives: To assess the reported prevalence of LVNC in different adult cohorts, taking into consideration the role of diagnostic criteria and imaging modalities. Methods: Systematic review and meta-analysis of studies reporting LVNC prevalence in adults. Studies were sourced from Pre-Medline, Medline and Embase, and assessed for eligibility according to a standardised proforma. Eligible studies provided a prevalence of LVNC in adult populations (≥ 12 years). Studies were assessed, and data extracted by two independent reviewers. Results: Fifty-three eligible studies documenting LVNC in 60 unique cohorts were included. The pooled prevalence estimates for LVNC were consistently higher amongst cohorts diagnosed on cardiac magnetic resonance (CMR) imaging (15%, n = 23) compared with echocardiogram (1%, n = 31). The prevalence of LVNC varied between disease and population representative cohorts. Athletic cohorts demonstrated high pooled prevalence estimates on echocardiogram (3%, n = 5) and CMR imaging (27%, n = 2). Conclusion: There is a higher prevalence of LVNC with newer imaging technologies, specifically cardiac magnetic resonance imaging, which identify LVNC changes more readily. The clinical significance of these findings remains unclear, however there is significant potential for overdiagnosis, overtreatment and unnecessary follow-up.


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Abstract P1.5 DISRUPTION OF RENAL TUBULAR MITOCHONDRIAL QUALITY CONTROL BY KCA3.1 IN DIABETIC NEPHROPATHY Chunling Huang1, Hao Yi1, Ying Shi1, Qinghua Cao1, Xin-Ming Chen1, Carol Pollock1 1. Renal Lab, Kolling Institute, University of Sydney, Royal North Shore Hospital , St Leonards, NSW 2065, Australia Background: Mitochondrial dysfunction is involved in the pathogenesis of diabetic nephropathy. Mitochondrial quality control is characterized by repairing of mitochondrial damage through mitophagy and fission/fusion. It has been shown that blockade of KCa3.1, a potassium channel, ameliorates diabetic renal fibrosis. However, the role of KCa3.1 in mitochondrial quality control is not yet known. Aim & Objectives: To identify the role of KCa3.1 in mitochondrial quality control in diabetic nephropathy. Methods: In vitro human proximal tubular cells (HK2 cells) transfected with scrambled siRNA or KCa3.1 siRNA were exposed to TGF-β1 for 48h. Mitochondrial function, mitochondrial ROS (mtROS) production were assessed. In vivo, diabetes was induced in KCa3.1+/+ and KCa3.1-/- mice by streptozotocin injection. The mitochondrial fusion related protein optic atrophy 1 (OPA1) and fission related protein dynamin-related protein (Drp1) as well as the autophagy marker LC3 were examined by western blotting in HK2 cells and mice kidneys. Results: The in vitro results showed that TGF-β1 significantly inhibited mitochondrial ATP production rate when compared to the controls, which were significantly reversed by KCa3.1 gene scilencing in HK2 cells. KCa3.1 gene silencing inhibited TGF-β1-induced significant increase in MitoSOX Red fluorescence in HK2 cells. TGF-β1 significantly decreased OPA1, increased Drp1 and LC3 expression in HK2 cells, which were reversed by KCa3.1 gene silencing. Consistently, KCa3.1 deficiency significantly reversed diabetes induced decreased OPA1 and increased Drp1 and LC3 expression in HK2 cells and diabetic KCa3.1-/- mice. Conclusion: KCa3.1 mediates dysregulation of mitochondrial quality control in diabetic nephropathy. Abstract P1.6 KINEMATIC VERSUS MECHANICAL ALIGNMENT IN PRIMARY TOTAL KNEE REPLACEMENT: A SYSTEMATIC REVIEW AND META-ANALYSIS Joshua Xu1, Jacob Cao1, Jason Luong1, Jonathan Negus1,2 1Sydney Medical School, The University of Sydney, NSW, Australia 2Department of Orthopaedics, Northern Beaches Hospital, Frenchs Forest, NSW, Australia Background: The two primary alignment methods in total knee replacement (TKR) are mechanical alignment (MA) and kinematic alignment (KA). MA restores the limb to a neutral alignment while KA restores the knee to its natural pre-arthritic alignment. There is discussion on the optimal knee alignment technique. Aim & Objectives: To compare the functional outcomes, radiological outcomes and complication rates of kinematic alignment (KA) and mechanical alignment (MA) for primary total knee replacement (TKR). Methods: Electronic searches were performed using five databases from their date of inception to January 2018. Relevant studies were identified, with data extracted and meta-analysed from the identified studies. Results: From the 8 studies included, there were 330 patients undergoing KA and 335 patients undergoing MA for TKR. We found that the postoperative Knee Society Function Score (P=0.004), Combined Knee Society Score (P=0.002) and flexion range of motion (P=0.01) was significantly better in the KA group when compared to the MA group. Also, the mean operation time (P=0.01) for the KA group was shorter than that of the MA group. However, there was no significant difference between the KA and MA groups in terms of radiological outcomes, complication rates (P=0.80) and reoperation rates (P=0.99). Conclusion: Our findings suggest that KA in TKR produces better functional outcomes than MA and has shorter operation times. However, there were no differences between both techniques regarding radiological outcomes and complication rates. Further research in larger sample populations are required to validate these findings and examine long-term outcomes.


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Abstract P1.7 BIOMARKERS OF AUTONOMIC BALANCE: A NEW HORIZON FOR PREVENTING MENTAL DISORDER AND IMPROVING RECOVERY AMONG CRASH SURVIVORS Ilaria Pozzato1, Ashley Craig1, Bamini Gopinath1, Yvonne Tran1, Michael Dinh2, Mark Gillett3, Ian D Cameron1 1 John Walsh Centre for Rehabilitation Research, Northern Clinical School, Faculty of Medicine and Health, The University of Sydney, Kolling Institute of Medical Research, Sydney, Australia. 2 Emergency Department, Royal Prince Alfred Hospital, Sydney, Australia. 3 Emergency Department, Royal North Shore Hospital, Sydney, Australia. Background: Psychological distress is an important well-known contributor to serious mental disorders and poorer recovery after road traffic injuries. Measurements of autonomic balance early after the crash may be a promising non-invasive tool to detect survivors’ ability to cope with stress. Aim & Objectives: This study aims to investigate (i) risk of psychological distress and poor recovery up to 12 months after a crash, (ii) ability of autonomic balance biomarkers in early detecting at-risk individuals, (iii) links between mental and physical health outcomes. Methods: This is a controlled prospective cohort study, where 120 mildly-to-moderately injured crash survivors presenting to two Emergency Departments in Sydney, will be assessed at 1, 3, 6 and 12 months of their injury. Outcomes are autonomic biomarkers, such as a heart function measure, called heart rate variability, and a combination of bio-psychosocial measures based on the WHO International Classification of Functioning. Comparisons to matched non-injured controls, undertaking the same longitudinal assessments, will also occur. Results: Ethical approvals (HREC/15/RPAH/423) and prospective registration (ACTRN 12616001445460) have been obtained. To date, ongoing recruitment counts 120 crash survivors and 80 controls. Conclusion: Findings will clarify whether autonomic biomarkers (perhaps in conjunction with psychosocial predictors) can assist in early identification of people who are vulnerable to developing psychopathology. If findings are positive, these are likely to improve management of traffic injuries in acute and general care settings and to enhance compensation processes, for a faster personal recovery, decreased medical and insurance costs, and lower rates of mental disorders among crash survivors. Abstract P 1.8 PROTEIN C LEVELS ON ADMISSION PREDICT BURNS PATIENT OUTCOME AND ITS TREND PARALLELS RECOVERY Ruilong Zhao1, Thomas Lang1, Rachel McGrath2, Meilang Xue1, Yan Lin1, Sue Smith3, Albert Kim2, Siobhan Fitzpatrick2, Nancy Huang2, Andrew Zimmerman1, Aruna Wijewardena2, Greg Fulcher2, John Vandervord2, Chris Jackson2 1 Sutton Laboratory, Kolling Institute at Royal North Shore Hospital, Sydney, NSW, Australia 2 Royal North Shore Hospital, Sydney, NSW, Australia 3 Raymond Purves Laboratory, Kolling Institute at Royal North Shore Hospital, Sydney, NSW, Australia Background: The natural course of partial-thickness burns is difficult to predict without robust markers to assist traditional evaluation. Aim & Objectives: We assessed the potential of: activated protein C (APC), a protease with pleiotropic beneficial properties; its zymogen, protein C (PC); and their receptor, endothelial protein C receptor (EPCR), to predict burns patient outcomes. Methods: We enrolled 88 severe burns patients in a single-centre prospective cohort study. Biopsies of normal and burned skin were collected from each patient and immunostained for PC and EPCR. Plasma was collected on admission (Day 0) and every three days, and analysed for PC, APC, soluble (s)EPCR, and inflammatory cytokines. Clinical data were recorded throughout admission. Results: Epidermal PC expression was reduced (p<0.01) while EPCR was increased (p<0.05) in burn-damaged compared to normal tissue. Correspondingly, Day 0 plasma PC levels were markedly reduced (p<0.0005), then recovering over time to a plateau by Day 9. Day 0 plasma PC levels predicted requirement of increased medical support during admission, even when adjusted for burn depth, size and inhalational injury (p=0.013; OR 0.879 [0.793-0.973]). APC increased from a nadir on Day 3, to Day 15. Plasma sEPCR and cytokines did not vary significantly over the study period. Conclusion: PC expression is decreased in burn-damaged tissue. Day 0 plasma PC is a better predictor of increased support during admission than burn just clinical characteristics alone. Although a validation study is required, plasma PC levels may be a novel marker for predicting outcomes in patients with severe burns.


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Abstract P1.9 EMBEDDING RESEARCH BIOBANKING INTO THE WORKFLOW OF A DIAGNOSTIC PATHOLOGY LABORATORY – THE ROYAL NORTH SHORE HOSPITAL EXPERIENCE Sam Yuen1,2, Ussha Pillai1, Nathan Hall2, Kimiko Blendell2, Cameron Turner2, Antony Kaufman2, David Nevell2, Mikaela Holmes1, Shannon Chan1, Jane Carpenter3, Verity Hodgkinson3,4, Roger Wilson3, Kathleen Phillips3, Simon Cooper3 and Deborah J Marsh1,5 1Hormones and Cancer Group, Kolling Institute, Royal North Shore Hospital (RNSH), St Leonards, NSW, Australia and University of Sydney, Sydney, NSW Australia 2Department of Anatomical Pathology, Royal North Shore Hospital, NSW Health Pathology, St Leonards, NSW, Australia 3NSW Health Statewide Biobank, NSW Health Pathology, Camperdown, NSW, Australia 4Sydney Medical School, The University of Sydney, Westmead, NSW, 2145, Australia 5School of Life Sciences, Faculty of Science, University of Technology Sydney, NSW, Australia Background: Tumour banking underpins translational cancer research. While tumour banks have existed for well over 20 years in NSW, the cost to diagnostic pathology laboratories of supporting them has not been established. Furthermore, the potential advantages of embedding tumour banking within routine workflows of diagnostic pathology laboratories have not been explored. Aim & Objectives: This project sought to establish a workflow pipeline for biobanking multiple tumour streams for research purposes within an Anatomical Pathology department. It has aimed to collect data to determine the impact of research biobanking on the time of staff employed in a diagnostic facility. Methods: This study was conducted as an 18 month collaboration between the Kolling Institute Tumour Bank and RNSH Anatomical Pathology (AP). It supported a Technical Officer position to focus on embedding a biobank workflow within RNSH’s AP department. Data was generated on procedures involved in the collection of research specimens, including the time various processes took and the staff involved. Results: Embedding this workflow has involved all facets of patient care, including theatre staff, surgeons, laboratory staff, registrars, pathologists and tumour bank officers. Engagement with all stakeholders has involved educational briefings, staff in-services, new protocols and guidelines. This project has elucidated numerous differences between medical research and clinical diagnostic environments in the context of workflows and objectives. Conclusion: The embedding of research focused workflows into the workflow of a diagnostic laboratory must address differences and engage in top down and bottom up directives in order to seamlessly integrate the two ideals. Abstract P1.10 CHEMICAL PROFILING OF EXHALED BREATH FROM CYSTIC FIBROSIS PATIENTS USING GC×GC-TOFMS Mohammad Asif Iqbal1, Maiken Ueland1, Katie D. Nizio1, Yang Song2, Raynuka Lazarus2, Lucy Kealley2, Jenny Bishop2, Peter Middleton2, Tapan Rai1, and Shari L. Forbes1 1School of Mathematical and Physical Sciences, University of Technology Sydney, NSW, Australia 2Cystic Fibrosis Group, The Westmead Institute for Medical Research, NSW, Australia Background: Chronic bacterial lung infections are the leading cause of death in patients with Cystic Fibrosis (CF). To date, sputum culture is the most common technique for the diagnosis of bacterial lung infections in adult CF patients. However, it requires several days or longer to obtain culture results. Therefore, a rapid diagnostic technique for bacterial lung infections would significantly improve CF healthcare. Aim & Objectives: The objective of this study was to investigate volatile organic compounds (VOCs) in exhaled breath samples from adult CF patients and to identify biomarkers specific to certain bacterial lung infections. Methods: In this study, breath samples were collected from both adult CF patients and healthy controls and analysed using a combination of solid phase microextraction (SPME) and comprehensive two-dimensional gas chromatography – time-of-flight mass spectrometry (GCxGC–TOFMS). In addition, VOCs in the headspace of corresponding CF sputum samples were also analysed using the same technique. The results were evaluated using principal component analysis (PCA) in order to assess the multivariate structure of the data. Results: The results demonstrate moderate variance among groups (e.g. CF sputum vs. control breath and CF breath vs. control breath). It was also possible to identify a set of potential biomarkers responsible for the variance between groups, however further data is required. Conclusion: The overall finding of this study indicates that the VOC profile obtained from exhaled breath allows distinction between groups (e.g. CF vs healthy controls). In addition, it provides important clues to the diagnosis of bacterial lung infections in adult CF patients.


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Abstract P1.11 CLINICAL SIGNIFICANCE OF KDM6A EXPRESSION IN MELANOMA

Gaya Punnia-Moorthy,1, Jason Madore2 , James Wilmott2, Richard Scolyer 2 , Justin Wong 3 , Peter Hersey 1, Jessamy Tiffen 1 1Melanoma Oncology and Immunology Group, Centenary Institute, NSW, Australia 2 Melanoma Institute Australia, NSW, Australia 3 Epigenetics and RNA biology, Centenary Institute, NSW, Australia Background: Melanoma is an aggressive form of skin cancer and the most common type of cancer in young adults. Australia has one of the highest incidence of melanoma in the world. Current treatments for metastatic melanoma are plagued by the resistance these melanomas develop against current targeted therapies or immunotherapies. Lysine demethylases (KDMs) are epigenetic enzymes that remove methyl groups from the amino acid lysine on histone proteins, which effects gene expression. One of these KDMs is KDM6A (also known as UTX), that removes methyl groups from histone 3, lysine number 27 (H3K27me3) inducing activation of gene expression. KDM6A has been reported to play an important role in the progression of bladder cancer, multiple myeloma, lung cancer, cervical cancer and pancreatic cancer however the role of KDM6A in melanoma is yet to be investigated. Aim & Objectives: To correlate KDM6A expression with clinical parameters and overall survival in a cohort of patients with melanoma Methods: In this study, we generated tissue microarrays (TMAs) and quantified KDM6A protein expression in 83 patients with cutaneous melanoma using immunohistochemistry (IHC) and correlated KDM6A expression with various clinical parameters including survival. Results: The results showed that 55% of patients had high KDM6A expression and that the median KDM6A IHC score in the cohort was 1.6. A score >1.6 was used to classify expression as ‘high’ where as a score </= 1.6 defined ‘low’ expression. KDM6A expression significantly correlated with disease free status, prior cancer diagnosis and specimen site. In addition, patients with high KDM6A expression lead to significant decreased overall survival compared to patients with low KDM6A. Conclusion: KDM6A expression significantly correlated with various clinical parameters and that high KDM6A expression had poorer overall survival. These results suggest that KDM6A has important clinical implications in melanoma and that targeting KDM6A in melanoma should be further investigated. Abstract P1.12 INFLAMMATION OF THE INFRAPATELLAR FAT PAD AND CLINICAL OUTCOMES OF THE KNEE: THE INTERLOCK STUDY PROTOCOL Hema Urban (Umapathy)1, Jillian P. Eyles1, David J. Hunter1, Kathryn Mills2 1. Institute of Bone and Joint Research, The Kolling Institute, University of Sydney, and Rheumatology Department, Royal North Shore Hospital, Sydney, New South Wales, Australia 2. Faculty of Medicine and Health Sciences, Macquarie University, Sydney, New South Wales, Australia Background: Anxiety is linked to increased pain sensitisation, but little is known about this association in osteoarthritis. Aim & Objectives: To investigate if pressure pain thresholds (PPT) are associated with anxiety in patients with patellofemoral osteoarthritis (PFOA). Methods: Participants with PFOA were recruited from the osteoarthritis chronic care program at the Royal North Shore Hospital. PPT was measured using a handheld algometer at 4 points around the patellar and on the extensor carpi radialis longus (ECRL) muscle of the non-dominant forearm in Newtons (N). Anxiety was measured using the Depression, Anxiety and Stress Scale questionnaire’s anxiety domain. The associations were examined using Pearson’s and Spearman’s correlation coefficients with Cohen’s standard used to determine the strength of the relationship (0.10-0.29 represented weak association; 0.30-0.49 moderate, and ≥0.50 strong association). Results: Baseline demographics showed only 27.2% of participants were classified with at least mild anxiety. All PPT points were significantly inter-correlated with strong associations (p = <0.001). Anxiety was weakly correlated with a reduced PPT measured at the Lateral site (r= -0.23, p=0.038) and the ECRL (-0.25, p=0.027) Conclusion: Lower local threshold in patients with increased anxiety suggest secondary hyperalgesia and the correlation of reduced ECRL PPTs with increasing anxiety suggests central sensitisation. These patients may also experience lower pain tolerance at the knee given the significant intercorrelation between local and distal thresholds. These findings highlight an important potential therapeutic pathway for PFOA patients; treatments to reduce anxiety may be useful in reducing pain sensitisation.


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Abstract P1.13 EFFECTS OF E-CIGARETTE ON HUMAN CORONARY ARTERY ENDOTHELIAL CELLS Michael Chhor1, Tara Nguyen1, Charles Cranfield1, Catherine Gorrie1, Kristine McGrath1 1 School of Life Sciences, University of Technology Sydney, NSW, Australia Background: Cardiovascular disease (CVD) is a leading cause of morbidity world-wide, with cigarette smoking being a major preventable risk factor. Smoking cessation can be difficult due to the addictive nature of cigarettes and the withdrawal symptoms associated with cessation. Electronic cigarettes (e-Cigs) have emerged as a popular smoking cessation device, however, the health effects surrounding the use of e-Cig remains unclear. Aim & Objectives: To investigate the (i) effects of e-Cig aerosol condensate (EAC) on human coronary artery endothelial cells (HCAEC) – key cells that plays a role in the initiation of atherosclerosis and (ii) effect of e-Cigs on the cardiovascular system using a mouse model of ‘vaping’. Methods: In vitro: HCAEC were exposed to EAC for 24hr before the effects of cell viability, oxidative stress and inflammation were assessed using an MTT assay, dichlorofluorescein assay and an ICAM-1 ELISA, respectively. A co-culture model using conditioned media of A549 epithelial lung cells was also used to assess the effects of EAC on HCAEC. In vivo: Following 12 weeks e-Cigs exposure, the aortic arch was collected from female Balb/C mice and stained for VCAM-1 using DAB staining. A one-way ANOVA with Bonferroni’s post-hoc test was used to determine significance for each assay. Results: In vitro, cell viability in HCAEC was decreased when exposed to EAC either directly or after exposure to A549 cells. Reactive oxygen species levels and ICAM-1 expression levels were shown to be increased in HCAEC when exposed to EAC directly. In vivo, VCAM-1 staining were increased in the aortic arches from animals exposed to e-Cigs. Conclusion: Together, these results provide evidence that e-Cigs can have an impact on the development of atherosclerosis and cardiovascular health. Therefore, caution should be taken before considering e-Cig as a cessation option. Further work is now needed to elucidate the full impact of e-Cig in CVD. Abstract P2.1 A NOVEL SCREENING TEST FOR COELIAC DISEASE USING GLIADIN COATED GOLD NANOPARTICLES Anantdeep Kaur 1, Michael Wallach2, Olga Shimoni1 1 Institute for Biomedical Materials and Devices, University of Technology Sydney, NSW, Australia 2 School of Life Sciences, University of Technology Sydney, NSW, Australia Background: Coeliac disease (CD) is an immune mediated disorder that damages the small intestine in genetically predisposed individuals. CD has high prevalence rates with up to 80% of affected individuals remaining undiagnosed, emphasising an immediate need for a valid serological or point-of-care test for early diagnosis of the disease. Aim & Objectives: Development of a novel screening test for CD based on the coating of gold nanoparticles with gliadin, the highly antigenic protein that induces CD. Methods: The binding of the hydrophobic gliadin protein on the surface of the gold nanoparticles (AuNPs) was successfully characterised. A serological assay was then developed in the form of a simple, single step screening test. The assay was tested on 30 patient serum samples in a blinded assessment and results were compared with the previously run serological and pathological tests on these patients. Results: Our data demonstrates that we can accurately detect the biomarker, anti-gliadin antibody; using a stable suspension of AuNPs coated with the whole gliadin protein in a single step assay. The results for the screening test on 30 patient non-haemolytic serum samples, were recorded after the visual examination of precipitate formation and the determination of a shift or change in absorbance values using a UV-Vis spectrophotometer. Based on the clinical sample analysis, it was demonstrated that a very high accuracy of (>95%) in distinguishing CD from non-CD patients was achieved. Conclusion: The one step assay indicates immense potential as a point-of-care test by eliminating multiple steps followed in existing serological tests. This test would be particularly useful in aiding the large-scale screening of the general population.


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Abstract P2.2 APPLICATION OF PHOSPHATIDYLETHANOL AS A BIOMARKER FOR ALCOHOL CONSUMPTION Van Long Nguyen1,2,3, Paul S. Haber3,4, Devanshi Seth3,4,5 1 NSW Health Pathology 2 Department of Chemical Pathology, Royal Prince Alfred Hospital, Missenden Road, Camperdown, NSW 2050, Australia 3 Faculty of Medicine, The University of Sydney, Sydney, NSW 2006, Australia 4 Drug Health Services, RPAH, Camperdown, NSW 2050, Australia 5 Centenary Institute, 93 Missenden Road, Camperdown, NSW 2050, Australia Background: Phosphatidylethanol (PEth) is a direct metabolite of ethanol. PEth can accumulate in red blood cells where it resides for approximately 4 weeks post alcohol consumption, unlike other indirect markers detected only within hours/days. Aim & Objectives: To validate PEth as a long-term biomarker for alcohol consumption. To determine whether the biomarker can identify different levels of drinking and its performance for ‘any’ drinking compared to routine alcohol biomarkers. Methods: PEth was analysed from whole blood samples using liquid chromatography tandem mass spectrometry. A mixed cohort of 133 individuals was tested. Correlations between self-reported alcohol consumption and biomarker levels were investigated. Receiver operator characteristic curves for four different drinking patterns were also examined for PEth. A comparison between PEth and common indirect alcohol biomarkers from routine blood testing were performed as well. Results: PEth displayed a positive correlation with self-reported alcohol consumption (r2 = 0.738). PEth displayed much higher discrimination for distinguishing between heavier forms of drinking (>40 g ethanol/day) (area under the curve (AUC) = 0.96) than compared to lower-moderate drinking levels (<40 g of ethanol/day) (AUC = 0.60). In the context of any alcohol consumption in the preceding month, PEth was the most sensitive (96%) and specific (88%) biomarker. This was compared to liver function tests, mean corpuscular volume and %carbohydrate deficient transferrin. Conclusion: PEth presents as a suitable longer-term biomarker for indications of alcohol consumption. PEth testing may be considered in settings to supplement an individual’s self-report. Abstract P2.3 SDHAF3 INTERACTS WITH SDHB: COULD AN SDHAF3 VARIANT PLAY A ROLE IN PHEOCHROMOCYTOMA AND PARAGANGLIOMA? Trisha Dwight,1,2 Un Na,3,4 Edward Kim,1,2 Ying Zhu,5 Anne Louise Richardson,1 Bruce G Robinson,1,2 Katherine M Tucker,6 Anthony J Gill,2,7,8 Diana E Benn,1,2 Roderick J Clifton-Bligh,1,2 Dennis R Winge3 1 Cancer Genetics, Hormones and Cancer Group, Kolling Institute of Medical Research, Royal North Shore Hospital, Sydney 2065, Australia 2 University of Sydney, Sydney 2006, Australia 3 Department of Medicine, University of Utah Health Sciences Center, Salt Lake City, Utah 84132, United States of America 4 Department of Biochemistry, University of Utah Health Sciences Center, Salt Lake City, Utah 84132, United States of America 5 Hunter New England Health, Royal North Shore Hospital, Sydney 2065, Australia 6Hereditary Cancer Service, Prince of Wales Hospital, Sydney 2031, Australia 7Department of Anatomical Pathology, Royal North Shore Hospital, Sydney 2065, Australia 8Northern Cancer Translational Research Unit, Royal North Shore Hospital, Sydney 2065, Australia Background: Succinate dehydrogenase assembly factors (SDHAF1-4) play a crucial role in maturation of SDH subunits and assembly of the functioning SDH complex as a whole. Methods and Results: Through use of massively parallel sequencing, we identified a variant in SDH assembly factor 3 (SDHAF3), c.157T>C (p.Phe53Leu) associated with increased prevalence in pheochromocytoma and paraganglioma (6.6%) compared to normal populations (1.2% [1000 Genomes], p=0.003; 2.1% [Exome Aggregation Consortium], p=0.0063). As this variant was deemed “probably damaging” to protein function (by in silico prediction tools), we explored functional consequences of the amino acid change (p.Phe53Leu). In yeast, introduction of SDHAF3 variant (p.Phe53Leu) into Sdh7 null yeast (ortholog of SDHAF3 in humans) resulted in impaired function, as observed by its failure to restore SDH activity when expressed in Sdh7 null yeast relative to WT SDHAF3. As SDHAF3 is involved in maturation of SDHB, we tested the functional impact of SDHAF3 c.157T>C and various clinically relevant SDHB mutations on this interaction. Our in vitro studies in human cells show that SDHAF3 interacts with SDHB (residues 46 and 242), with impaired interaction observed in the presence of the SDHAF3 c.157T>C variant. Conclusions: Our studies reveal novel insights into the biogenesis of SDH, uncovering a vital interaction between SDHAF3 and SDHB. We have shown that SDHAF3 interacts directly with SDHB (residue 242 being key to this interaction), and that a variant in SDHAF3 (c.157T>C [p.Phe53Leu]) may be more prevalent in individuals with pheochromocytoma and/or paraganglioma, and is hypomorphic via impaired interaction with SDHB.


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Abstract P2.4 KILLING ME SOFTLY: MITOCHONDRIAL-MEDIATED NEUROTOXIC DEATH MECHANISMS OF BUTYLONE, PENTYLONE AND 3,4-METHYLENEDIOXYPYROVALERONE (MDPV) IN SH-SY5Y CELLS Huey Sze Leong1,2, Morgan Philp1, Martin Simone2, Paul Kenneth Witting2, Shanlin Fu1 1Centre for Forensic Science, School of Mathematical and Physical Sciences, University of Technology Sydney, NSW, Australia 2Discipline of Pathology, School of Medical Sciences, The University of Sydney, NSW, Australia Background: Illicit use of synthetic cathinones (SCs) butylone, pentylone and 3,4-methylenedioxypyrovalerone (MDPV) has resulted in a number of fatalities. The precise mechanism of drug toxicity is, however, unclear. Aim & Objectives: This paucity of understanding raises an interest to investigate in-vitro toxicity and its mechanistic pathways. Methods: Human neuronal SH-SY5Y cells were differentiated to a dopaminergic phenotype using retinoic acid and 12-O-tetradecanoylphorbol-13-acetate. Differentiated cells were treated with each drug (1 to 10mM) to assess its cytotoxicity. To define underlying neurotoxicity mechanisms; mitochondrial bioenergetics impairment, intracellular calcium (Ca2+) and cell death pathways measurements were evaluated at two dosages, EC15 (butylone 4.67, pentylone 3.05 and MDPV 2.60mM) and EC40 (butylone 5.91, pentylone 4.06 and MDPV 3.43mM). Results: After 24h treatment, all three SCs displayed dose-dependent cytotoxicity, with MDPV being most potent, followed by pentylone and butylone. Significant (p< 0.0001 vs control) decreased mitochondrial respiration parameters followed by markedly depleted adenosine triphosphate (ATP) contents signified mitochondrial dysfunction as the crucial factor in the onset of SCs-induced neurotoxicity. Significant alteration in Ca2+ homeostasis finally lead to mitochondrial-apoptotic cell death, as indicated by the substantial activation of caspases 3, and 7. Conclusion: Understanding the relationship between underlying neurotoxic mechanisms and related clinical manifestation is imperative in the management and treatment of SCs addiction. The potential clinical relevance for this study include identification that SCs trigger neuronal apoptosis and stimulate increased intracellular Ca2+ and mitochondrial dysfunction leading to a loss in ATP, information that may be useful in understanding the detrimental impact of these drugs. Abstract P2.5 MEMBERS OF THE CHLORIDE INTRACELLULAR ION CHANNEL PROTEIN FAMILY DEMONSTRATE PROTEIN DEGLUTATHIONYLATION ACTIVITY Hala M Ali1, Khondker R Hossain1, Claudia D’Amario, Lele Jiang1, Stella M Valenzuela1, 2

1 School of Life Sciences, University of Technology Sydney, Sydney, NSW 2007, Australia, 2Institute for Biomedical Materials and Devices, University of Technology Sydney, Sydney, NSW, 2007Australia Background: The chloride intracellular ion channel (CLICs) proteins are atypical ion channel proteins, as they are principally soluble proteins that also demonstrate enzymatic activity. Human CLIC proteins have been found in most tissues and cells. Our group was the first to empirically demonstrate that CLIC family members have intrinsic enzymatic activity. Furthermore, we now have evidence that the expression of CLIC proteins by E. coli bacterial cells provides them with increased tolerance to oxidative stress. Aim: Our current study demonstrates for the first time that CLICs possess significant deglutathionylating activity. Method: We used a model peptide substrate in conjunction with a real-time fluorescence-based method. Results: Mutating the active site cysteine residue in CLIC1 (Cys24) largely eliminated its deglutathionylating activity. Furthermore, mutating either one of two highly conserved charged residues, arginine 29 (R29A) or lysine 37 (K37A) in CLIC1, where both residues are located in the putative transmembrane domain, and are also within the vicinity of the substrate binding site, resulted in a reduction of its deglutathionylation activity. We also now have indirect evidence of CLIC1 dependent de/glutathionylation of cellular proteins detected in cultured cell lines that are over-expressing CLIC1. Conclusion: The discovery that CLIC proteins participate in the glutathionylation cycle and would likely be targeting specific proteins, could explain the association of CLICs with a diverse range of clinical disorders and provide a novel therapeutic target.


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Abstract P2.6 TEMPORAL PROFILING OF PLASMINOGEN ACTIVATORS IN DOPAMINERGIC DIFFERENTIATED NEURONAL CELLS EXPOSED TO LIPOPOLYSACCHARIDE (LPS) Aram Niaz1, Jocelyn Karunia1, Mawj Mandwie1, Ghaith Al-Badri1, Alessandro Castorina1,2 1 School of Life Sciences, University of Technology Sydney, NSW, Australia 2 Discipline of Anatomy & Histology, School of Medical Science, The University of Sydney, NSW, Australia Background: Parkinson’s disease (PD) is an incurable neurodegenerative condition that can severely affect the physical, mental, and emotional well-being of humans. Diagnosing PD at the earliest stages can improve treatment outcomes, but suitable biomarkers are currently lacking. Aim & Objectives: The goal of this project is to assess the temporal expression pattern of plasminogen activators (PAs) and compare their expression profile with that of a panel of pro-inflammatory cytokines in dopaminergic differentiated neuronal cells exposed to LPS, to mimic an inflammatory insult. Methods: Human neuroblastoma SH-SY5Y cell lines were differentiated into dopaminergic neurons using 12-O-tetradecanoylphorbol-13-acetate (TPA). Cells were exposed to LPS either for short-term (up to 24h) or long-term (up to 10 DIV), and the expression of PAs, pro-inflammatory markers and that of different intracellular cascades was assessed by real-time qPCR or Western blot. Results: We identified some degree of linearity between the temporal expression of members of the PA system and that of canonical pro-inflammatory cytokines. Additional studies revealed that the expression of some PA members correlated with the activation of the PI3K/Akt/cAMP-responsive element binding protein (CREB) cascade. Conclusion: Taken together, the outcomes of this study suggest that certain members of the PA system hold the promise to be considered as valid candidate biomarkers of neuroinflammation in inflamed dopaminergic cells, and suggest that the PI3K/Akt/CREB pathway may be responsible for PAs gene induction following LPS exposure. Abstract P2.7 NR4A1 DEFINES AN ANTI-INFLAMMATORY SENESCENT CELL POPULATION Sorour Jarrah1, Paul R Coleman1, Jia Li1, Yang Zhao1, Kaka Ting1, Mathew A Vadas1, Jennifer R Gamble1 1. Centre for the Endothelium, Vascular Biology Program, Centenary Institute. Background: Cellular senescence is characterised by a permanent halt in the cell cycle, but where the cells remain viable and metabolically active. It is essential for development and wound healing, but can also result in chronic disease, should these cells not be cleared through its elaboration and stimulation of a pro-inflammatory milieu. Endothelial cell senescence contributes to cardiovascular disease and is also linked to cancer progression. However, senescent endothelial cells can also acquire an anti-inflammatory phenotype, as we have recently shown. This suggests that senescence in the vasculature can have a dual action both to stimulate (the pro-inflammatory senescent) and inhibit (the anti-inflammatory senescent) inflammation. However, there is a lack of appropriate markers to identify the two phenotypes of senescent endothelial cells limiting our capacity to understand the impact of these in disease initiation and progression. Aims: We have recently shown using RNAseq transcriptomic analysis, that NR4A1, a member of the Orphan Nuclear Receptor family subfamily 4 group A member 1 (NR4A1) is highly up-regulated in the anti-inflammatory senescent endothelial cells. NR4A1 is a transcriptional co-factor that inhibits inflammation. Here, we explored the link between the anti-inflammatory phenotype of senescent endothelial cells and up-regulation of the expression of NR4A1 in this population Method: Depletion of NR4A1 reverted the inflammatory senescent phenotype of the endothelial cells, from anti- to pro-inflammatory without affecting the number of senescent endothelial cells. Conclusion: NR4A1 is a new marker for identification of the unique anti-inflammatory senescent endothelial cell phenotype and will allow investigations into the temporal expression of this cell type in age-associated diseases.


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Abstract P2.8 EFFECT OF LONG-TERM POLYPHARMACY AND THE DRUG BURDEN INDEX (DBI) ON CARDIOVASCULAR FUNCTIONS AND CARDIAC FIBROSIS IN AGED MICE Trang Tran1,2, John Mach1,2, Gizem Gemikonakli1,2, Alexander Widiapradja1,2, Scott P Levick1,2, Susan Howlett3, Rafael de Cabo4, David G Le Couteur2,5 & Sarah N Hilmer1,2 1 Laboratory of Ageing and Pharmacology, Kolling Institute, Royal North Shore Hospital, Sydney, NSW, Australia 2 Nothern Clinical School, University of Sydney, NSW, Australia 3 Dalhousie University, Halifax, Canada 4 Translational Gerontology Branch, National Institute on Aging, Maryland, USA 5 ANZAC Research Institute, Sydney, NSW, Australia Background: Polypharmacy (use of ≥ 5 medications) and increasing Drug Burden Index (DBI: cumulative exposure to anticholinergic/sedative drugs) impair function in older adults. Preclinical studies can provide a mechanistic understanding of these exposures on organ function. Aim & Objectives: To evaluate whether chronic polypharmacy or monotherapy, with increasing DBI and deprescribing (medication cessation) affect cardiovascular functions and histology in aged mice. Methods: 12-month-old male C57BL/6 mice received control chow or medicated feed containing polypharmacy regimens of Zero-DBI (simvastatin, metoprolol, omeprazole, paracetamol, irbesartan), Low-DBI (simvastatin, metoprolol, omeprazole, paracetamol, citalopram), High-DBI (simvastatin, metoprolol, oxybutynin, oxycodone, citalopram) or monotherapy (medications from High-DBI regimen administered independently), all at therapeutic doses. At 21-months, animals were re-randomised to continue treatment or be deprescribed. Blood pressure (BP) and rotarod performance were assessed every 3-months. Hearts were collected at 26-months for collagen quantification. Results: At 21-months, compared to control, systolic and diastolic BP decreased in Zero-DBI, Low-DBI, simvastatin and metoprolol treated mice (P<0.05) but not in High-DBI (has metoprolol and simvastatin) group (P>0.1). At 21-24-months, compared to control and High-DBI, rotarod latency-to-fall, adjusted for weight and cohort, increased in metoprolol group (P<0.05). Neither BP nor rotarod endurance differed significantly with each treatment compared to its deprescribed group. Preliminary results (n=5) indicate that compared to control, High-DBI increased myocardial collagen (P<0.05) while Zero-DBI, Low-DBI, metoprolol and simvastatin showed no significant effect. Conclusion: Chronic treatment with this High-DBI polypharmacy regimen may impair therapeutic effects of antihypertensives and increase myocardial collagen. Future studies will continue to explore morphological changes involved. Abstract P2.9 THE POTENTIAL CHONDRO-PROTECTIVE EFFECTS OF 26S PROTEASOME INHIBITORS IN AN IN VITRO CARTILAGE EXPLANT MODEL Ying Liu1,3, Cindy C Shu1,3, Varshini Ravi1,3, Shihani Stoner1,3, Susan M Smith1,3, Anthony W Ashton2,3, Christopher B Little1,3 1 Raymond Purves Bone and Joint Research Laboratories, Institute of Bone and Joint Research 2 Perinatal Medicine Research 3 Kolling Institute, Royal North Shore Hospital, Faculty of Medical Health, University of Sydney Background: 26S proteasome inhibitors (PIs) have demonstrated capability in preventing NFkB activation, hence are candidate therapeutics for protecting injury or inflammation-induced cartilage damage. Aim & Objectives: We investigate the relative chondro-protective efficacy of three PIs (MG132, Bortezomib and Carfilzomib) and their mechanisms of action. Methods: Full-thickness ovine articular cartilage was cultured in serum-free DMEM ± 10ng/ml IL1β ± 20μg/ml MG132, 1μM Bortezomib or 20nM Carfilzomib. At 24/96hrs, conditioned media and cartilage were collected for sulphated-glycosaminoglycan (GAG) quantification, histology, and gene expression (COL2A1, ACAN, MMP3, MMP13, ADAMTS4, ADAMTS5, TIMP1, TIMP3). N=6/treatment/time point. Results: At 24hrs PIs did not alter GAG release compared to control, while MG132 and Bortezomib (but not Carfilzomib) reduced IL1β-induced GAG release. At 96hrs Bortezomib alone increased GAG release, while only MG132 inhibited IL1β-induced loss. The pattern of GAG release was reflected in the loss of cartilage proteoglycan histochemical staining. MG132 and Bortezomib (not Carfilzomib) inhibited expression of COL2A1, ACAN, TIMP1/3, increased ADAMTS4/5 and MMP3, but decreased MMP13. While some temporal differences were seen, in the presence of IL1β MG132 and Bortezomib generally augmented COL2A1, ACAN, TIMP1/3 inhibition, and inhibited ADAMTS4 and MMP13. Although Carfilzomib did not modify IL1β-induced GAG release, it resulted in less ACAN inhibition and greater ADAMTS4/5 inhibition than the other PIs, but did not modify MMP13. Conclusion: The three PIs investigated conferred distinct effects against IL1β-induced cartilage aggrecanolysis in vitro. However effects on the expression of key cartilage matrix homeostasis genes did not readily explain the protective effect of MG132 on preventing proteoglycan loss. Abstract P2.10


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PATIENT-DERIVED ENDOTHELIAL PROGENITOR CELLS DISPLAY SEX-SPECIFIC DIFFERENCE IN ANGIOGENESIS, AND DISEASE-RELATED DIFFERENCE IN REDOX SIGNALLING Christine Yu1, Kristen Bubb1, Gemma Figtree1 1. Cardiothoracic and Vascular Health Research, Kolling Institute, University of Sydney, Royal North Shore Hospital, ST Leonards, NSW, 2065 Background: Increasing age amplifies the production of free radicals which contributes to cardiovascular disease, where women are uniquely vulnerable. Endothelial cells (EPCs) play a vital role in regeneration post MI, which led to our interest in the potential mechanistic insight they can provide. Aim & Objectives: To investigate signalling pathways relevant to vascular function and disease within EPCs extracted from patients undergoing clinically indicated CT coronary angiography (CTCA). Methods: Patients in the BIOHEART study at North Shore Private Hospital had background data and blood samples taken prior to CTCA scan. Peripheral Blood mononuclear cells (PBMCs) were cultured in 0.1% gelatin coated T25 flasks and protein lysate was collected and used in immunoblotting, determining angiogenic and redox signalling molecules. Cells were also seeded onto a tube formation assay (Matrigel). Results: The average expression of Nox2 in EPCs in patients with confirmed coronary disease was almost double versus healthy recruits (2.0±0.37 vs. 1.02±0.12, p=0.02 by unpaired t-test, n=14-13). We observed a trend towards increased tube formation between patients with disease versus without (2.45±0.35 1.64±0.24, p=0.06 by unpaired t-test, n=15-15). EPCs from females with coronary disease showed significantly greater tubule formation than EPCs from healthy females (3.22±0.60 1.3±0.34, p<0.05 by unpaired t-test, n=6-4). Conclusion: The significantly increased expression of Nox2 in EPCs of patients with coronary atherosclerotic disease, suggests that EPCs reflect dysregulated vascular redox signalling. With an unfavourable risk of death in females, the trend towards increased angiogenic capabilities may reflect a compensatory response to coronary atherosclerosis. Abstract P2.11 GENOMIC SURVEILLANCE OF KLEBSIELLA SPP. ISOLATED FROM PATIENTS WITH BACTERAEMIA: INSIGHTS FROM A LOCAL COHORT STUDY. Kyle Catalan1, Michael Liu2, Thomas Gottlieb3, Elaine Cheong3, John Merlino3, Steven Djordjevic1 and Piklu Roy Chowdhury1,2 1 The i3 institute, University of Technology Sydney, Ultimo, NSW 2007, Australia 2Department of Primary Industries, Elizabeth Macarthur Agriculture Institute, [email protected] 3 Concord Repatriation General Hospital, Microbiology and Infectious Diseases, NSW Health Pathology, Concord NSW, Sydney Background: Klebsiella species are included as pathogens of clinical interest in the “Global Antimicrobial Resistance Surveillance System” (2015) and are considered high-risk drug resistant micro-organisms. A database linking isolate metadata with genome sequence and phylogeny would enhance understanding of the epidemiology of this important group of pathogens. Aim & Objectives: There is limited knowledge on the population structures and resistance gene carriage of Klebsiella spp. circulating in Australia. Here we conducted whole genomic sequencing (WGS) of Klebsiella isolates from patients with bacteraemia at Concord Hospital, Sydney between 2013-2016, to create a baseline database. Methods: Genomes of 129 Klebsiella (105 pneumoniae and 24 oxytoca) isolates collected between 2013 and 2016 from Concord Repatriation General hospital were sequenced using Illumina technology. The short-read sequences were assembled using Shovill and analysed in-silico using a range of open-source software, including pubMLST, PlasmidFinder, ResFinder, ISFinder, ARIBA (Antimicrobial Resistance Identification By Assembly), RAST (Rapid Annotation using Subsystem Technology) and Mauve. Results: We identified 62 different K. pneumoniae sequence types (ST) and 9 K. oxytoca STs. Notably, we identified 19 novel STs of pathogenic Klebsiella (8 pneumoniae and 11 oxytoca) not reported previously. Fourteen of 129 (10.9%) isolates were multidrug-resistant, of which 5 were resistant to extended-spectrum beta-lactam antibiotics. Two hyper-virulent lineages with ST25 and ST86 were identified and these had specific genomic hotspots that could facilitate the rapid evolution of complex resistance phenotypic variants. Conclusion: We identified both novel lineages of Klebsiella spp. and two hyper-virulent lineages within the bacteraemia cohort which is worth monitoring.


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Abstract P2.12 FOOD AND INFLAMMATION Rachel Shepherd1 1 School of Medical and Molecular Biosciences, University of Technology Sydney, NSW, Australia Background: It has been reported that some foods have inflammatory or anti-inflammatory properties. Aim & Objectives: To examine the evidence for inflammatory and anti-inflammatory effects and clarify the biological mechanisms for these effects. Methods: The evidence from published research was surveyed and potential biological mechanisms investigated. Results: There are a large number of scientific publications reporting inflammatory and anti-inflammatory effects for common food components. Many of these studies have quantified increases and decreases in a variety of different serum inflammatory response agents (CRP, cytokines, interleukins etc). However not all subjects exhibited similar serum effects with inconsistencies generally ascribed to genetic differences, with little evidential support. Also there was limited corroborating support for some of the effects. The proposed mechanisms for these effects were diverse and largely under-investigated. Conclusion: There is evidence that a variety of compounds commonly found in foods can influence inflammatory markers in serum. The mechanisms for these effects are less clear. Abstract P2.13 USING GENOMICS AND PROTEOMICS TO UNDERSTAND THE ANTIBIOTIC RESISTANCE CAPABILITIES OF AN ISOLATE Stephanie Town1, Steven P Djordjevic2, Matthew P Padula1 1 School of Life Sciences, University of Technology Sydney, NSW, Australia 2 Infection, Immunity and Innovation (i3) Institute, University of Technology Sydney, NSW, Australia Background: Recently, next-generation genomic sequencing has shown potential as an accurate predictor of phenotype and for understanding genes conferring antibiotic resistance (AMR) in bacterial communities. In the clinic, antibiotic disc assays can provide a visual indication of the type and strength of AMR in an isolate. Aim & Objectives: Our research aims to understand the AMR capabilities of an isolate on a molecular level through examination of its genome and proteome, with and without antibiotic challenge. This research seeks to connect AMR genes to gene end-products on the proteoform-level (peptides, proteins, post-translational modifications), which has yet to be experimentally shown. Methods: Long- and short-read genomic sequencing and assembly was conducted in-house on multi-drug resistant E. coli isolates to ascertain the presence of AMR-related genes and demonstrate phenotypic effectiveness for related-AMR. A shotgun proteomics pipeline using LC-MS/MS was used to generate data on proteome changes with and without antibiotic challenge. PEAKS Studio was used to analyse data, and UniProt and STRING databases indicated cloud- and crowd-based evaluations. Results: Several interesting proteins related to AMR were upregulated, despite no antibiotic challenge, including proteins previously annotated as hypothetical and multi-drug resistance proteins. Conclusion: This research establishes that proteomic techniques validate the potential for genomic sequencing to replace current clinical tests and provide more specificity in selecting the right antibiotic in the clinic. Our research findings are signif icant for being the first to experimentally link an AMR-related gene to the AMR-related protein using a systems biology approach that provides evidence that the genotype and the phenotype do differ.


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Abstract P2.14 INDIRECT PHOTOBIOMODULATION: A NOVEL TREATMENT FOR NEURODEGENERATION? Ji Yeon Kim1234, Thomas Ashhurst56, Matt Dun7, Daniel Johnstone34 1 Faculty of Medicine, University of Queensland, Herston, Australia 2 University of Queensland Centre for Clinical Research, Brisbane, Australia 3Bosch Institute, University of Sydney 4Discipline of Physiology, University of Sydney 5 Discipline of Pathology, University of Sydney 6 Sydney Cytometry, Centenary Institute, Sydney, Australia 7School of Biomedical Sciences and Pharmacy, University of Newcastle Background: Photobiomodulation (PBM) – treatment of tissue with low-intensity near-infrared light (λ=600–1100nm) – induces neuroprotection in animal models, however clinical translation is limited by light attenuation across the human skull. Treating peripheral tissues with ‘indirect/remote’ PBM also induces neuroprotection, and shows promise for clinical translation. Aim & Objectives: Assess efficacy and mechanism of remote PBM Methods: APP/PS1 mice (transgenic Alzheimer’s model) were treated with 50mW/cm2 670nm remote PBM for 3min/day, 3d/wk, for 8wks (8J/cm2/3min treatment). Brains were immunohistochemically stained for markers of pathology, including amyloid plaques. BALB/c and C57BL/6 mice were treated with remote PBM (4J/cm2/90sec treatment/day). Serum collected after 2d of treatment was subjected to ELISAs for target molecules FGF-21 or SDF-1a, or to proteomics analysis. Bone marrow collected after 10d of treatment was assessed by flow cytometry for changes in stem cell populations. Results: Indirect PBM did not significantly impact amyloid plaque load or density. There was no significant difference in FGF-21 or SDF-1a levels between treated and sham groups. Following proteomics analysis of serum (n=5/group), 842 proteins were identified with high confidence (p<0.01). Of these, 209 had a sham vs. treatment fold change of >1.25 or <0.8, and 15 had a fold change of >1.5 or <0.67. Flow cytometry revealed proliferation of a subset of mesenchymal (CD117hi) and haematopoietic (MPP1) stem cells. Conclusion: Systemic response to remote PBM appears multifaceted, with cellular and gene expression components playing a part. Other immunohistochemical markers may reveal more regarding efficacy. Increased understanding of this novel treatment may contribute to clinical translation. Abstract P2.15 DEVELOPMENT OF NEW THERAPY TO COMBAT TUBERCULOSIS INDUCED INFLAMMATION Giang HB Le1, Jessica Pedersen 1,2, AkaneTanaka1, Sheila Donnelly 1, Nilesh J Bokil1, Bernadette M Saunders 1,2 1School of Life Science, University of Technology Sydney, NSW, Australia 2Centenary Institute and Sydney Medical School, The University of Sydney, Sydney, NSW, Australia Background: Tuberculosis (TB) is a major health problem worldwide. While curable, TB infection often results in a strong pro-inflammatory response that can cause lasting lung damage. Helminth infections are often associated with the release of proteins with anti-inflammatory activity. Aims and Objectives: This study is examining the capacity of the anti-inflammatory helminth protein- helminth defence molecule (FhHDM-1) to treat mycobacteria induced inflammation. Methods: To examine this, primary human macrophages were treated with FhHDM-1 either before, simultaneously with, or after infection with Mycobacterium bovis Bacille Calmette-Guérin (BCG. The effect of FhHDM-1 on activation of macrophage pro-inflammatory function and bacterial survival was assessed. Results: Treatment with FhHDM-1 significantly inhibited the secretion of the pro-inflammatory proteins TNF, IL-6 and MCP-1. Interestingly, simultaneous treatment was not as effective as pre- or post- treatment in reducing secretion of these pro-inflammatory proteins. Interestingly no significant change was observed in the expression of mRNA for these markers upon treatment with FhHDM-1. Macrophages treatment with FhHDM-1, were able to control bacterial growth as efficiently as untreated macrophages. Conclusion: These data show that FhHDM-1 has an ability to inhibit the pro-inflammatory response induced by BCG infection. Furthermore, the peptide does not affect the capacity of the macrophages to control bacterial growth. These data suggest that further studies investigating the potential of FhHDM-1 to act as an adjunct therapy to reduce the excessive inflammation associated with tuberculosis infection are warranted.


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AWARDS AND PRIZES Awards and prizes include:

Honours Students: • Best Oral Presentation by an Honours Student ($500 cash prize) • Best Oral Presentation by an Honours Student – runner up ($300 cash prize) • Popular choice award - $100 Young Investigators (PhD students only): • Best Oral Presentation by a Young Investigator ($500 cash prize + $1,000 travel grant) • Best Oral Presentation by a Young Investigator – runner up ($300 cash prize + $700 travel grant) • Popular choice award - $100 Early Career Researchers (within 10 years post-PhD): • Best Oral Presentation by an Early Career Researcher ($500 cash prize + $1,000 travel grant) • Best Oral Presentation by an Early Career Researcher – runner up ($300 cash prize + $700 travel

grant) • Popular choice award - $100

Josephine Anderson Award (technical officer / research assistant only): • Josephine Anderson Award for Best Oral Presentation ($500 cash prize) • Josephine Anderson Award for Best Poster ($100 cash prize) Open to all: • Best Poster Awards (5 x $100 cash prizes) • Popular choice award – 3 x $50 Note: Travel grants can be claimed back (with valid receipts) from the conference committee within one calendar year from the date of the 2018 New Horizons conference. The travel grant can be used to support any costs related to airfare, accommodation and registration for presenting (oral or poster) at a national or international conference.

New Horizons 2018 | Innovative science with impact 47

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EPPENDORF SOUTH PACIFIC PTY. LTD. Contact: Rhea Cornely Address: Unit 4 112 Talavera Rd

North Ryde NSW 2113 Tel: 02 9889 5000 Email: [email protected] Website: www.eppendorf.com/AU-en

MEDIKANE Contact: Malcolm Ball Address: 123 Epping Road, Macquarie Park NSW 2113 Tel: 1300 889 962 Email: [email protected] Website: www.medikane.com.au

PHARMAXIS LTD Contact: Wolfgang Jarolimek Address: 20 Rodborough Road, Frenchs Forest NSW 2086 Tel: 02 9454 7200 Email: [email protected] Website: www.pharmaxis.com.au


http://www.beckman.com.au/


http://www.bio-rad.com/


http://www.bio-strategy.com.au/


https://biotools.com.au/


http://www.bmglabtech.com/


http://www.eppendorf.com/AU-en


http://www.medikane.com.au/


http://www.pharmaxis.com.au/


SPONSOR & EXHIBITOR INFORMATION

SAPPHIRE BIOSCIENCE Contact: Katie Chan Address: Unit G1/44-70 Rosehill Street, Redfern NSW 2016 Tel: 02 9698 2022 Email: [email protected] Website: www.sapphirebioscience.com


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kolling research institute 19th 20th november 2018€¦ · new horizons 2018 | innovative science...

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