List of research projects available for prospective graduate students Category: Cardiovascular Disease Department of Biochemistry Principal Investigator

Project Title with a brief description

Dr Jiang Jianming [email protected]

Identification and Dissection of the Regulatory Pathways For Heart Regeneration and Cardiovascular Disease Cardiovascular disease is a leading cause of death and disability worldwide. Heart failure (HF) occurs as a consequence of heart muscle damage. However, the adult mammalian heart, including the human heart, has limited potential to generate new muscle following injury and the burden of persisting decreased heart function may eventually lead to HF. Development of regenerative therapeutic strategies to reverse the progression of HF is an urgent and unmet clinical need. Recent research indicates regenerative potential of mammalian heart exists but dissipates shortly after birth. It is generally believed that the potential of heart regeneration is correlated with cardiomyocyte proliferative capacity. Interest in reinstating the generative potential of adult mammalian heart has led to exploration of the mechanisms underlying cardiomyocyte exit from their cell cycle to permanent arrest during the perinatal period. Our research interests focus on identifying and dissecting the regulatory pathways for heart regeneration using cutting-edge and integrated approaches in single cell RNAseq analysis, in vivo gene modulation, stem cell reprograming and high-throughput pharmacological and genetic screening. Our research will not only uncover novel regulators and pathways for reinstating regenerative potential in adult mammalian hearts, but also provide novel targets for drug discovery and lead to better approaches for treatment or prevention of cardiovascular disease.

Department of Medicine Principal Investigator

Project Title with a brief description

A/Prof Lee Chi-Hang, Ronald [email protected]

The aim of this randomized, open-label, clinical trial is to determine the impact of Sleep Study Guided Multidisciplinary Therapy (SGMT, i.e. continuous positive airway pressure and behavioral therapy) for obstructive sleep apnea (OSA)-in sub-acute phase of acute coronary syndrome on cardiovascular outcomes. We hypothesize that SGMT will result in a lower (1) plasma NT-pro BNP level, and (2) 10-year risk of cardiovascular mortality based on the European SCORE algorithm, when compared with Standard Therapy. OSA is being recognized as an emerging cardiac risk factor and prognostic marker. We have reported that OSA was a prevalent, as

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well as an independent predictor of plaque burden and adverse outcomes in patients presenting with acute coronary syndrome. In this clinical trial, which is a continuation of my research and publication trajectory, 180 patients presenting with acute coronary syndrome will be randomly assigned to SGMT (n=90) or Standard Therapy (n=90) groups. Both groups will receive guideline-mandated treatment for acute coronary syndrome. Those assigned to SGMT will undergo an overnight sleep study. Those found to have OSA will attend the SGMT clinic run by a multidisciplinary team at baseline, 1-, 3-, and 6-month. Advice on continuous positive airway pressure and behavioral therapy (weight loss, exercise, positional therapy, abstinence of alcohol and sleeping pills) will be given. The primary endpoint is plasma NT-pro BNP concentration in 6month follow-up. The secondary endpoint is 10-year risk of cardiovascular mortality based on European SCORE algorithm which includes age, sex, smoking status, systolic blood pressure, and serum total cholesterol or total/HDL-cholesterol ratio. In our aging population with an increasing prevalence of obesity, OSA will potentially become an increasingly important contributor to cardiovascular disease. Leveraging the collective expertise of a term of cardiologists and sleep physicians, our work will benefit society by advancing our understanding on the cardiovascular benefits of screening for and treating OSA. A/Prof Ling Lieng Hsi [email protected]

Maternal vascular status and influence on fetal and child development This research is part of the birth cohort study “Growing Up in Singapore Towards healthy Outcome” (GUSTO) of the TCR flagship programme “Developmental Origins: Singapore” (DevOS). Prenatal and perinatal factors determine the subsequent growth of the child. This project aims to determine a) cardiac and vascular adaptation, and their interaction during pregnancy. b) patterns of subclinical vascular disease in preconception women, and their relation to cardiovascular biomarkers c) if maternal vascular structure and function, which is influenced by premature atherosclerosis and collagen/elastin degradation, has any influence on the physical and metabolic characteristics of newborns and their subsequent development. A preconception cohort will be studied with several established techniques to assess structural atherosclerosis (carotid intima-medial thickness), arterial stiffness (carotid-femoral pulse wave velocity) and cardiac function (echocardiography) and ventriculo-arterial coupling. These measures will be repeated during pregnancy, at the height of cardiovascular stress, and post-partum. The student will have opportunity to: a) learn technical skills needed in conducting cardiac and vascular testing

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b) correlate these findings in preconception women with prevailing cardiometabolic milieu c) determine how the stress of pregnancy modifies these indexes d) ascertain if abnormal vascular structure affects fetal and newborn outcomes and later development. A/Prof Mark Chan [email protected]

Improving outcomes in acute Myocardial infarction through reversal of early and Late cardiac remodelling. (IMMACULATE study) 1. Multicentre registry of patients hospitalised for acute myocardial infarction (AMI) with serial cardiac imaging and blood sampling to identify biomarkers and therapeutic targets for post-MI ventricular remodelling and heart failure.

2. Registry-based randomised trial comparing outcomes of ventricular remodelling and hemodynamic stress among patients with AMI and elevated NT-pro-B-type natriuretic peptide receiving comprehensive telemedicine-guided post-MI treatment (remote intensive management) vs. standard non-telemedicine guided treatment

A/Prof Pipin Kojodjojo [email protected]

Artificial Intelligence Algorithms for Risk Prediction of Cardiac Adverse Outcome The detailed electrical signatures of cardiac function are predictive of future events. Because of the precision of electrical measurement, these waveforms can form the basis for training neural nets related to artificial intelligence training algorithms. The project will involve characterizing large datasets involving complex signals to predict outcome in cardiovascular patients. In one specific application, electrical patterns will be used to predict onset of a condition called ‘atrial fibrillation’, in a randomized, controlled trial. The student will operate at the interface of advanced computing, with links to the National Supercomputing Centre, scientists and specialist physicians The student will also work in a team with full ethics approval for patient data access to create prediction algorithms for improved risk determination.

Dr Roshni R. Singaraja [email protected] [email protected]

Updated in Jan 2018

The roles of novel genes in Diabetes and Cardiovascular Diseases Diabetes is a leading cause of morbidity and mortality in Singapore, as well as globally, and Singapore has the second-highest proportion of diabetics among developed nations, after correcting for age. Common genes and pathways connect diabetes with Cardiovascular Disease (CVD), which is the leading cause of death worldwide. Both Diabetes and CVD are complex disorders, caused by multiple genes and/or their interactions with environmental factors. Our group utilizes human genetic and genomic approaches to novel genetic target discovery, as well as human clinical studies, mouse models, and in vitro iPS and cell culture models to validate and decipher the roles of these novel genes in disease pathology. The overall goal of our several projects is to Page 3

identify and characterize novel genes affecting Diabetes and CVD, with a goal of eventual therapeutic development for these diseases. Project 1: Characterization of the role of CYP8B1 in cardiometabolic diseases Using human genetic studies, we have identified a novel gene, CYP8B1, with a potential role in lipid and glucose metabolism. We aim to further characterize the role of this gene utilizing iPS-cell derived hepatocyte and beta-cell models, as well as animal models, in order to derive mechanistic insight into the impact of this gene in cardiometabolic diseases. Project 2: Characterization of the role of ABCA8 in Diabetes Utilizing Mendelian genetic studies in human family cohorts, we have identified ABCA8 as a novel gene linked to both Diabetes and cardiovascular diseases. We aim to decipher the role of ABCA8 in these diseases utilizing human studies, mouse models, and in vitro iPS and cell culture models. Project 3: Characterization of the role of novel genes in lipid and glucose metabolism Using a deep sequencing approach in a human cohort, we have identified genes associated with lipid levels. Genetic studies in humans suggest roles for these genes in lipid/glucose metabolism. We aim to use in vivo CRISPR based approaches to validate the role of these genes in mouse models, as well in ex vivo cell culture models. Department of Obstetrics & Gynaecology Principal Investigator

Project Title with a brief description

Prof Ariff Bongso [email protected]

The therapeutic potential of stem cells from the Wharton’s jelly of the human umbilical cord

A/Prof Fong Chui Yee [email protected]

Updated in Jan 2018

We have isolated and studied a novel primitive stem cell population from the Wharton’s jelly of the human umbilical cord (hWJSC) that is gaining increasing popularity. In fact the International Society for Cellular Therapy (ISCT) in their recent newsletter has called hWJSCs ‘a novel source for tomorrow’s cellular therapy’. hWJSCs are noncontroversial and can be harvested painlessly in abundance from discarded human umbilical cords. They are proliferative with short population doubling times, making it possible to expand their numbers after short periods in culture. They resemble hBMMSCs but with some added unique characteristics. They have their own signature of mesenchymal markers (CD10+, CD13+, CD29+, CD44+, CD90+, CD14-, CD33-, CD56-, CD31-, CD34-, CD45-), their stemness properties are maintained over long periods in culture, and their frequency of colony-forming units is much higher than that of hBMMSCs, . Several groups have reported differentiation of Page 4

hWJSCs into bone [30] cartilage, adipocytes, hepatocytes, neurons, pancreatic islet-like cells [38], endothelial cells [39] and skeletal muscle cells. They are tolerated in preclinical xenograft models without the need for immunosuppression therapy, express HLA-G, MHC class I antigens and possess immunosuppressive properties in splenocyte proliferation and mixed lymphocyte reaction assays.They express lower levels of human leukocyte antigen-DR than their bone marrow counterparts and thus exhibit low immunogenicity in clinical applications. Safety studies showed that hWJSCs do not transform to tumour-associated fibroblasts unlike hBMMSCs, are non-toxic and do not induce tumorigenesis in laboratory animals and cynomolgus monkeys [54]. The engraftment, safety and regenerative ability of hWJSCs were demonstrated in xenograft animal models and recently in several human clinical trials. Interestingly, a recent pilot study showed that hematopoiesis was enhanced after co-transplantation of cord blood cells with hWJSCs but the authors claimed that more studies were warranted and the exact mechanisms as to how hWJSCs enhanced hematopoiesis required elucidation. Furthermore, besides the improved engraftment, it is not known whether the hWJSCs targeted and destroyed residual renegade lymphoma or leukemic cells. This proposal aims to find other novel potentials of the Wharton’s jelly stem cells for cell-based therapy. Prof Yong Eu Leong [email protected] Dr Susan LOGAN [email protected]

Integrated Women’s Health Program (IWHP) Singapore is globally one of the most rapidly aging societies. With improvements in health care, life expectancy in females has increased from 68 years to 85 years over the past 45 years. In 2013, women aged >45 years comprised 40% of Singapore’s female population &, as such, would expect to spend half of their lives in the menopausal transition & menopause. Many apparently well midlife Singaporean women attend gynecology for routine health screens and care. Currently this care is specialty centric but with a recognition that this age group face multiple health issues including problems relating to menopausal symptoms, bone health and osteoporosis, sexual health, cancer, cardiovascular disease, emotional health, mobility, balance & muscle strength. Many are unrecognized & unmet and all directly influence each other. Improving health care delivery and outcomes depends on systematically establishing the prevalence of health conditions and ultimately health-care needs. In addition, previous research has primarily focused on men. Gynaecologists are ideally placed to bring the focus to women & introduce individualized and innovative healthcare solutions for the prevention and timely treatment of chronic conditions such as hip fractures, cardiovascular disease/stroke, dementia, cancer, and emotional and healthy relationships. The IWHP consists of five elements: 1. A cross sectional study to assess the prevalence & correlates of

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health conditions and risk factors across multiple systems (cardiovascular, mental, musculo-skeletal, sexual and gynaecological) among a clinical population of women ages 45-69 years in Singapore. The assessment tool includes a questionnaire (electronic in 9/2015), biophysical measurements, blood sampling, short physical performance battery & whole body (bone, muscle, fat) DXA. a. To use this cross sectional information to compare the prevalence of health conditions in the IWHP to that of previously published findings from samples of women living in the United States & globally b. To use the fasted blood samples to explore novel biomarkers in relation to disease prediction 2. To integrate research findings within a treatment setting. Participants will be provided results from their research assessments and will be given referrals to internal resources. This model of care will be termed the Integrated Women’s Health Program (IWHP). 3. Using the IWHP as scaffolding for recruitment of eligible patients, it is proposed to initiate interventional clinical trials after identifying specific needs 4. To undertake a longitudinal assessment of this mid-life female cohort Department of Paediatrics Principal Investigator

Project Title with a brief description

A/Prof Heng Chew Kiat [email protected]

Research Topic: Genetics of coronary artery disease Coronary artery disease (CAD) is the leading cause of death and disability in many developed countries and is the number two killer in Singapore. According to the World Health Organization, 23.6 million deaths each year by 2030 are projected to be due to cardiovascular diseases. CAD is a complex disease that is caused by multiple genetic and environmental factors as well as their interactions. There are several ongoing projects being conducted in our Cardiovascular Genetics Lab. The common objectives of these projects are to determine the contributing genetic factors to CAD and to develop methods to better predict CAD risk. We have 3 ongoing projects available for graduate students. 1. Genome-wide association studies (GWAS) of coronary artery disease and its associated risk factors This study aims to identify susceptibility genes for CAD and myocardial infarction in the Singaporean population. Other risk factor phenotypes investigated include lipid profile, blood pressures and inflammatory markers. We also participate in both regional and international consortia such as Asian Genetic Epidemiology Network (AGEN) and Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE). Students wishing to work on this project are required to have a good grasp of biostatistics.

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2. Investigation of a newly identified protein, androgendependent tissue factor pathway inhibitor regulating protein (ADTRP), for its role in coronary artery disease Recently, a GWAS in the Chinese population has identified rs6903956 within the C6orf105 gene (now known as androgendependent tissue factor pathway inhibitor “TFPI” regulating protein or ADTRP) on chromosome 6p24.1 as a novel susceptibility locus for CAD. ADTRP is a protein encoded by C6orf105 that regulates TFPI expression. TFPI is the major inhibitor of tissue factor-factor VIIa– dependent FXa generation. Hence, ADTRP can be considered a protective factor of CAD. As it is a newly discovered protein, very little is known about its characteristics. This provides ample scope for investigation. The specific aims of this study are: 1. To determine which of the two known isoforms of ADTRP is the predominant form through the use of Human Coronary Artery Endothelial Cell (HCAEC) line. 2. To determine the changes in ADTRP in response to androgen in HCAEC. 3. To select meaningful single nucleotide polymorphisms for genotyping to determine if any are significantly associated with CAD in the Singaporean population. 4. To conduct genetic variant screening in ADTRP gene The methods employed in this study include cell culture, PCR, qRTPCR, genotyping, bioinformatics and statistical analyses. 3. Identification of rare variants with strong effects on early onset myocardial infarction There is an increasing trend of early onset of myocardial infarction (MI) in patients who are below 40 years. A proportion of these do not have strong conventional risk factors such as smoking, hypertension and diabetes. In fact some of the patients with early onset of MI lead physically active lives and do not appear to have any conventional risk factors except for being male. It is therefore of interest to investigate the genetic factors underlying this early onset phenotype. Genome-wide association studies (GWAS) have thus far identified genetic variants that may predispose individuals to MI. However, these are mostly variants that are common in the population and are hence unlikely to account for the small subset of patients who had MI at a young age. We aim to carry out exome sequencing using next generation sequencing (NGS) technology for MI patients 45 years comprised 40% of Singapore’s female population &, as such, would expect to spend half of their lives in the menopausal transition & menopause. Many apparently well midlife Singaporean women attend gynecology for routine health screens and care. Currently this care is specialty centric but with a recognition that this age group face multiple health issues including problems relating to menopausal symptoms, bone health and osteoporosis, sexual health, cancer, cardiovascular disease, emotional health, mobility, balance & muscle strength. Many are unrecognized & unmet and all directly influence each other. Improving health care delivery and outcomes depends on systematically establishing the prevalence of health conditions and ultimately health-care needs. In addition, previous research has primarily focused on men. Gynaecologists are ideally placed to bring the focus to women & introduce individualized and innovative healthcare solutions for the prevention and timely treatment of chronic conditions such as hip fractures, cardiovascular disease/stroke, dementia, cancer, and emotional and healthy relationships. The IWHP consists of five elements: 1. A cross sectional study to assess the prevalence & correlates of health conditions and risk factors across multiple systems (cardiovascular, mental, musculo-skeletal, sexual and gynaecological) among a clinical population of women ages 45-69 years in Singapore. The assessment tool includes a questionnaire (electronic in 9/2015), biophysical measurements, blood sampling, short physical performance battery & whole body (bone, muscle, fat) DXA. a. To use this cross sectional information to compare the prevalence of health conditions in the IWHP to that of previously published findings from samples of women living in the United States & globally b. To use the fasted blood samples to explore novel biomarkers in relation to disease prediction 2. To integrate research findings within a treatment setting. Participants will be provided results from their research assessments and will be given referrals to internal resources. This model of care will be termed the Integrated Women’s Health Program (IWHP). 3. Using the IWHP as scaffolding for recruitment of eligible patients, it is proposed to initiate interventional clinical trials after identifying specific needs 4. To undertake a longitudinal assessment of this mid-life female cohort

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Department of Paediatrics Principal Investigator

Project Title with a brief description

Prof Dario CAMPANA [email protected]

Cell therapy of cancer The available projects focus on developing novel immunotherapies for cancer by expanding and genetically-modifying immune cells, such as T lymphocytes, natural killer cells and monocytes. The research relies on methods developed in our laboratory, some of which have already been translated into clinical application. The research involves a variety of techniques including cell culture, gene cloning, vector design and flow cytometry.

Department of Physiology Principal Investigator

Project Title with a brief description

A/Prof Christopher Beng-Ti ANG Sialylation and Tumor Cell Invasion in Glioblastoma [email protected] Brain tumors are among the most devastating of cancers, with a mean survival period of just 15 months upon. Currently, clinicians rely on mainly histology to guide treatment decisions. In line with international efforts, we recently showed that resistance to current chemotherapeutic drugs and tumor cell invasiveness can be attributed to a self-renewing, cellular fraction within the tumor mass. This tumor heterogeneity is the reason that brain tumors are notoriously difficult to treat. We further showed that patient genetic profiles can be mapped to such gene activation profiles enriched for stem-like properties. Our proposal here will focus on one such important gene activation program, regulated by a sialylation enzyme, ST3Gal1. We recently demonstrated that ST3Gal1 expression marks a persistent cellular fraction, and is highly enriched in mesenchymal molecular features, typical of aggressive and recurrent tumors. Our orthotopic animal model established from patient cells further demonstrated that ST3Gal1 knockdown abrogated tumor-initiating and -sustaining capability. Moving forward, we will focus on signaling mechanisms that mediate ST3Gal1 activity. Our lab has expertise to interrogate patient clinical databases, and we identified several methylation hallmarks that potentially regulate ST3Gal1. This is a significant List of research projects available for prospective graduate students Updated in Aug 2015 Page 49 endeavor as DNA methylation accounts for up to 70% of tumor profiles. We will test such methylation hallmarks in vitro and in animal models. Our lab deals with several techniques such as primary cell culture, biochemical and molecular biology approaches and flow cytometry. Histology forms a significant thrust of our analyses, along with mouse models that recapitulate the patient heterogeneous spectrum. Our study is unique as we rely on patient genomic content to guide our bench experiments. The successful candidate will be exposed to cross-disciplinary approaches to Updated in Jan 2018

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address clinically important questions in glioma biology.

A/Prof Herbert Schwarz [email protected]

Development of a CD137L chimeric antigen receptor for DC immunotherapy Chimeric antigen receptors (CAR) are a breakthrough technology and among the most promising new cancer therapies. This project will extend the use of CARs by employing them for inducing an anticancer immune response. For this we will use a novel discovery and technology developed by us, which is the generation of dendritic cells by CD137 ligand signaling (CD137L-DC) that are several fold more potent than classical DC. Monocytes transfected to express a CAR which upon binding to a tumor antigen induces differentiation of the monocytes to CD137L-DC will prime tumor-specific T cells that eliminate tumor cells (Figure 1). Figure 1: Schematic depiction of the construction of a CD137L-CAR specific for Her2, a tumor associated antigen, to be used for generating CD137L-DC that will induce an immune response against breast cancer.

Exploration of CD137 ligand induced myelopoiesis for treatment of Acute Myeloid Leukemia CD137L is expressed on the surface of hematopoietic stem and progenitor cells (HSPC) and of antigen presenting cells (APC), and CD137L is capable of transmitting signals into HSPC and APC, a process referred to as reverse signaling. A main effect of CD137L signaling is inducing myeloid differentiation. We are exploring this activity of CD137L for its usefulness in treating Acute Myeloid Leukemia (AML). AML is due to hematopoietic progenitor cells failing to complete the myeloid differentiation pathway. Our recent data demonstrate that recombinant CD137 protein, which engages CD137L on AML cells, induces their differentiation. This is expected to reduce the aggressiveness of AML. We aim to complement these in vitro studies with in vivo studies in murine AML models. Further, we will characterize the CD137L signaling cascade in order to identify potential small molecule targets for AML differentiation therapy.

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A/Prof Lina Lim [email protected]

Interplay between immune cells and cancer cells to drive tumorigenesis The molecular mechanisms involved in initiation, promotion and progression of cancer are still unclear; inflammation is thought to play a major role in all aspects of tumorigenesis. Similarly, the immune system may play a large role in the sustenance or regression of the tumor, and the understanding of the role of the immune response towards tumors will be important in the development of innovative anti-tumor therapies. The strong association between macrophages and tumor cells as well as the high incidences of leukocyte infiltration in solid tumors have made tumor-associated macrophages (TAMs) to be seen as the key to tumor progression and are considered to be putative therapeutic targets for cancer treatment (Sica et al, 2008). Significant progress has been made to understand how TAMs induce cancer growth and how tumors induce macrophage polarization to TAM. However the signals with which TAMs provide to tumors to promote tumor metastasis is unclear. ~30% of breast cancer metastasizes to the brain and bone in HER2+ and Triple negative breast cancer, resulting in untreatable circumstances. Therefore, it is of vital importance that new insights into the mechanisms of metastasis to the brain and bone are needed.

Prof Shazib Pervaiz [email protected]

Redox Regulation carcinogenesis

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Over the years, our work has highlighted the critical role of an altered redox metabolism on cell survival and death signaling in cancer cells. Using a variety of model systems such as drug-induced apoptosis, receptor mediated death signaling, and oncogene-induced cell survival, we demonstrated that the intracellular ratio between the two main reactive oxygen species (ROS), superoxide and hydrogen peroxide (O2 - :H2O2), determines cancer cell response to death signals; a tilt in favor of superoxide promotes cell survival whereas an increase in hydrogen peroxide favors death execution via activation of the death promoting protein Bax. Of note, we have highlighted a novel biological activity of Bcl-2 by providing experimental evidence linking Bcl-2-induced increase in mitochondrial superoxide levels to the anti-apoptotic activity of Bcl-2. Interestingly, an elevated O2 :H2O2 ratio induced by either pharmacological inhibition (DDC) or gene knockdown of Cu/Zn SOD resulted in an increase in phosphorylation of Bcl-2, specifically at Ser70 (S70), and this site specific phosphorylation of Bcl-2 enhanced the anti-apoptotic activity of Bcl-2, thereby rendering cancer cells resistant to chemotherapyinduced apoptosis. To that end, we have uncovered a novel mechanism in which an increase in intracellular O2 - endows cancer cells with a survival advantage via tyrosine nitration-mediated detachment of B56δ from PP2A catalytic core, and the eventual accumulation of S70 phosphorylated Bcl-2 with potent anti-apoptotic activity. The mechanism of redox modulation of PP2A in the context of carcinogenesis is under investigation. Updated in Jan 2018

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Specific targeting of mutant K-Ras expressing cancers via Aktdependent ROS production This project involves investigating the molecualr mechanism(s) underlying the activity of a novel small molecule compound against mutant Kras expressing human cancers. Identification of a novel protein TMTC2 in death receptor sensitization of human cancers. Our preliminary studies indicate that ligation of the death receptors DR4 and DR5 (TRAIL R1 and TRAIL R2) upregulates the expression of a novel transmembrane protein TMTC2. Upregulation of TMTC2 increases death receptor sensitivity while its gene knockdown inhibited TRAIL-mediated cell death in human nasopharyngeal carcinoma cell lines. The sub-cellular localization of TMTC2 appears to place it at the ER, however this has not been validated in more than one cell lines. Notably, TMTC2 induction appears to be under the influence of intracellular reactive nitrogen species (RNS). The mechanism of induction, protein structure and localization as well as the death sensitizing activity of TMTC2 is under study. Identification of surrogate death signaling cisplatinresistant human cancer cells.

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Platinum-based compounds are the main line of treatment for a number of clinical cancers. However, development of drug resistance remains a therapeutic challenge. We have generated cisplatinresistant clones from human lung and ovarian carcinoma cell lines. These cells become resistant to most chemotherapeutic drugs, thus suggesting a MDR phenotype. Although, the mechanism of this drug resistance is not clearly understood, we have made a remarkable observation during the course of these studies, i.e that cells that become resistant to cisplatin become highly sensitive to death receptormediated apoptosis. The mechanism of activation of this surrogate death signaling in the context of cisplatin resistance is under investigation. Mechanism of statin-induced apoptosis in cancer cells. We recently demonstrated that the cholesterol lowering drug, simvastatin, induces apoptosis in human colorectal and breast carcinoma cells. We identified a critical role for intracellular ROS and downstream JNK activation in statin-induced apoptosis. Interestingly, we showed a massive induction and activation of Rho family of proteins, Rac1, Rho and cdc42 in statin treated cells, upstream of ROS production. Of note, the Rho family of proteins is prenylated despite their localization in the cytoplasm as opposed to the plasma membrane. The mechanism of this non-membrane dependent prenylation as well as ROS production in this model is under investigation. Updated in Jan 2018

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A/Prof Celestial T. Yap To investigate the roles of cytoskeletal proteins in oncogenic [email protected] signaling and cell survival Gelsolin (GSN) is an actin-associated cytoskeletal protein involved in binding and severing actin filaments, thus controlling cytoskeletal turnover in response to cellular signals (eg. migration, apoptosis). Gelsolin has also emerged as a new player in pathways regulating signaling and gene transcription. We previously uncovered a novel role of gelsolin in promoting invasion of colorectal tumour cells by enhancing extracellular matrix breakdown, through the upregulation of urokinase plasminogen activator (uPa). Recently we have also uncovered gelsolin’s roles in oncogenic growth factor pathways in gastric cancer (work accepted for publication). We aim to ascertain if specific oncogenic signaling pathways interact with gelsolin to influence cancer cell behaviour. Our analysis of microarray data from a variety of human cancer tissues and in vitro work on cell lines showed a correlation between gelsolin levels and various oncogenic signaling pathway modulators. These suggest possible involvement of gelsolin in regulating oncogenic signaling. The specific signaling pathways identified to have potential interactions with gelsolin are known to induce aggressive tumour cell behavior, including invasion, epithelial-mesenchymal transformation and resistance to cell death induced by cellular stresses. Gelsolin levels influence the expression of target genes controlled by oncogenic signaling. In addition, we found that induction of specific oncogenic signaling in breast cancer cells also increased the expression of gelsolin, suggesting that the activities of gelsolin may be recruited by certain oncogenic signaling pathways in a feedback loop to enhance their target effects. Collectively, these data suggest novel regulatory roles of gelsolin in signaling pathways that promote cancer progression. We will investigate in depth the interactions between gelsolin and the specific signaling pathways identified by our laboratory, as well as examine the significance of these interactions on cancer cell behavior and clinical outcomes. To investigate the roles of secreted miRNAs in modulating the tumour microenvironment Novel miRNAs secreted by human gastric cancers have been identified by our group, which distinctly differentiate cancers from non –cancer tissues. These secreted miRNAs are hypothesized to act on neighbouring cells in the tumour microenvironent to influence cancer spread. The project will investigate how subsets of the secreted miRNAs impact on cancer behavior and alter the tumour microenvironment.

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Department of Surgery Principal Investigator

Project Title with a brief description

A/Prof Edmund Chiong [email protected]

Gene Polymorphisms and Response to Bacillus CalmetteGuerin Therapy for Bladder Cancer Patients Hypothesis: The gold standard immunotherapy for NMIBC is BCG immunotherapy and our present studies indicate that NRAMP1 gene polymorphisms correlate with response to BCG response Singapore NMIBC patients. In this study, we seek to validate our results in a larger patient cohort, and hypothesize that NRAMP1 gene polymorphisms may predict cancer recurrence and response to BCG therapy in a more heterogenous population. We also seek to compare its predictive value with other known gene polymorphisms that have been suggested to predict for BCG response or bladder cancer risk. Aims: 1) To determine if NRAMP1 gene polymorphisms can predict the clinical outcome of Asian and European NMIBC patients who have undergone BCG therapy, and, 2) to compare its predictive value with other known gene polymorphisms. Methodology: A multi-centre study in Singapore, Hong Kong and Switzerland will be conducted. NMIBC patients (n=300) who had transurethral resection (TUR) and received at least a minimum 6 once-weekly plus 3 once-a-week intravesical BCG for EORTC intermediate or high risk disease, will be retrospectively recruited. Genomic DNA isolated from blood will genotyped and verified using high resolution melting (HRM) analysis and DNA sequence analysis respectively. Patient demographic, initial disease characteristics, vaccination status, intravesical BCG therapy details, and clinical outcome data will be collected and analyzed using Cox regression methods and Kaplan–Meier analyses. Clinical significance: Our proposed work may help to determine and confirm the importance and possibly identify ethnic differences with regards to the effect of NRAMP1 gene polymorphisms on BCG response. These results will further elucidate our understanding on the predictive value of NRAMP1 gene polymorphisms, and possibly aid in the decision for alternate therapy thus sparing the patient from the side effects and cost of BCG therapy.

Targeted Drug Delivery for Non-muscle invasive Bladder Cancer A/Prof Edmund Chiong [email protected]

Updated in Jan 2018

Intravesical chemotherapy for bladder cancer is highly promising for reducing recurrence rate in the short-term, but fails in the long-term, due to the short residence time and low penetration of the drug within the bladder. Cytotoxicity associated with administered chemotherapeutic agents limits higher doses for increased efficacy. Page 23

Thus, it is a challenge to achieve an ideal intravesical agent that can target and penetrate into tumor cells and be highly effective with minimal toxicity. The proposed work aims to develop targeting strategies with different drug formulations for more effective drug delivery to bladder tumors. We propose to design peptides to target fibroblast growth receptor 3 (FGFR3), epidermal growth factor receptor (EGFR) and CD47 on human bladder tumor cells. Screening experiments will be first carried out by evaluating the ability of the proposed peptide to bind two or more cell lines which overexpress FGFR3, EGFR or CD47. A number of intravesical drug delivery systems such as prodrugs and drug-nanoparticulate formulations will be developed by our collaborators in engineering. The drug-containing nanoparticles will be conjugated with tumor-targeting peptides to achieve effective targeting of bladder tumors. Our task is to further evaluate the targeted penetration in tumor cells and the corresponding therapeutic efficacy of the drug-nanoparticulate formulations conjugated with/without targeting peptides in a mouse orthotopic model. The use of fluorescence tagged peptide conjugated carriers as possible bladder cancer diagnostic tool will also be evaluated ex vivo with tumor biopsy samples and cell pellets from patients’ urine.

Category: Metabolic Diseases Research Department of Biochemistry Principal Investigator

Project Title with a brief description

Dr Nguyen Nam Long [email protected]

Discovery of physiological ligands for a lipid transporter In continuation with the discovery of a LPC/lipid transporter and the importance of DHA uptake for brain growth and CNS functions (Nguyen et al, Nature 2014, Nguyen et al., Nature Genetics 2015), my lab has identified a novel lipid transporter. This novel transporter protein is expressed in non-CNS tissues consistent with the strong peripheral phenotypes exhibited in knockout mice. Students will characterize the physiological functions of this novel transporter by analysis of the knockout phenotypes. This in vivo information will be important to design experiments to identify the physiological ligands of this transporter in vitro. New knowledge and information generated by understanding of the fundamental molecular and physiological functions of this transporter will facilitate the development of interventions for lipid-related diseases such as, cardiovascular diseases, diabetes and obesity. Students with strong interests in biomedical research are expected to have direct supervisions by the PI and expected to gain knowledge and various skills in genes, protein, lipid biochemistry, and cell culture assays as well as the physiological characterizations of knockout mice. Molecular and physiological characterization of a novel antilipid peroxidation enzyme in protection of kidney from diabetic nephropathy

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My lab has strong interests in identification and characterization of nutritionally regulated genes with a focus on lipid metabolism. We identified a few novel genes which are upregulated during fasting. Using our generated antibodies, we show that one of the gene candidates is highly expressed in polarized cells in the proximal tubules in kidney where 60% reabsorption of nutrients occurs and in microvilli enterocytes in small intestine which is the primary site for absorption of dietary nutrients. The preliminary data suggest the physiological roles of this novel protein in anti-oxidation of lipids. Students will develop assays to unravel the biochemical activity of this enzyme. In addition, students will learn to develop mouse models to study the physiological functions of this protein. We aim at developing a diabetic nephropathy model to better understanding pathology of the disease. Students are expected to gain knowledge and skills in protein and lipid biochemistry, cell culture assays as well as the physiological characterizations of knockout mice.

Adjunct Assistant Professor TEO Human in vitro models for studying diabetes disease Kee Keong Adrian mechanisms [email protected]; [email protected] Human pancreatic beta cell failure is the ultimate root cause of all forms of diabetes. Due to the limited availability of human islets and the lack of human models to study human beta cell biology, rodent islets and beta cell lines have traditionally been used as surrogate models. The recent discovery of human pluripotent stem cells (hPSCs) and the generation of human beta cell lines now provide wonderful opportunities to study human beta cells in vitro. Our projects involve the use of hPSC-derived pancreatic cells, human islets and the human beta cell line to investigate yet-elusive diabetes disease mechanisms. Methodologies such as the culture and differentiation of hPSCs into pancreatic cells, culture of human islets and beta cell lines, gene knock down and overexpression, QPCR, immunostaining, flow cytometry, manipulation of signaling pathways, glucose stimulation assays and high throughput technologies will be employed. The student is expected to have a deep interest in Stem Cells and Diabetes, be highly motivated, read the literature extensively, gain indepth knowledge on the research topic and master cell culture cum molecular biology techniques quickly. There will be intensive direct mentorship and a structured programme within the laboratory to bring the student up to speed on the topic of Stem Cells and Diabetes. This project is expected to gear the student towards a research career in Stem Cells, Diabetes and Metabolism. REFERENCES 1) Teo, K.K.A.*, Gupta, M.K., Doria, A., and Kulkarni, R.N.* (2015). Updated in Jan 2018

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Dissecting diabetes/metabolic disease mechanisms using pluripotent stem cells and genome editing tools. Mol Metab, accepted. *Corresponding authors 2) Teo, K.K.A., Tsuneyoshi, N., Hoon, S., Tan, E.-K., Stanton, L.W., Wright, C.V., and Dunn, N.R. (2015). PDX1 binds and represses hepatic genes to ensure robust pancreatic commitment in differentiating human embryonic stem cells. Stem Cell Reports 4, 113. 3) Valdez, I.A.#, Teo, K.K.A.#*, and Kulkarni, R.N.* (2015). Cellular stress drives pancreatic plasticity. Sci. Transl. Med. 7, 273ps2. #First authors *Corresponding authors 4) Teo, K.K.A.*, Valdez, I.A., Dirice, E., and Kulkarni, R.N.* (2014). Comparable generation of Activin-induced definitive endoderm via additive Wnt or BMP signalling in absence of serum. Stem Cell Reports 3, 5-14. *Corresponding authors 5) Teo, K.K.A., Wagers, A.J., and Kulkarni, R.N. (2013). New opportunities: harnessing induced pluripotency for discovery in diabetes and metabolism. Cell Metabolism 18, 775-791. 6) Teo, K.K.A., Windmueller, R., Johansson, B.B., Dirice, E., Njolstad, P.R., Tjora, E., Raeder, H., and Kulkarni, R.N. (2013). Derivation of human induced pluripotent stem cells from patients with maturity onset diabetes of the young. J Biol Chem. 288, 5353-5356. 7) Teo, K.K.A., and Vallier, L. (2010). Emerging use of stem cells in regenerative medicine. Biochem. J. 428, 11-23. A/Prof Yew Wen Shan [email protected]

Current Synthetic Enzymology research interest focuses on protein engineering and biocatalysis, with emphasis on using structural and mechanistic enzymological knowledge to develop therapeutics. We are involved in the following research areas: 1. Defining Synthetic Enzymology as an enabling platform for the

2.

3. 4. 5. 6.

Updated in Jan 2018

Construction of Next-Generation Synthetic Biology solutions for Pharma and Consumer Businesses. Using Synthetic Enzymology to discover and develop novel therapeutic biomolecules, in the areas of infectious diseases, metabolic disorders, cancer and aging. Developing anti-virulence Quorum-Based Technology for use in the biomedical industry. Using Synthetic Lixiviant Enzymology for Biomining of Electronic Wastes for Environmental Sustainability. Development of Lead Compounds Targeting Enzymes involved in Metabolic Cancer. Using Synthetic Enzymology for the Construction of Therapeutic (Probiotic) Cells for the Treatment of Metabolic Diseases and Page 26

Infectious Diseases. Department of Medicine Principal Investigator

Project Title with a brief description

Dr Roshni R. Singaraja [email protected] [email protected]

The roles of novel genes in Diabetes and Cardiovascular Diseases Diabetes is a leading cause of morbidity and mortality in Singapore, as well as globally, and Singapore has the second-highest proportion of diabetics among developed nations, after correcting for age. Common genes and pathways connect diabetes with Cardiovascular Disease (CVD), which is the leading cause of death worldwide. Both Diabetes and CVD are complex disorders, caused by multiple genes and/or their interactions with environmental factors. Our group utilizes human genetic and genomic approaches to novel genetic target discovery, as well as human clinical studies, mouse models, and in vitro iPS and cell culture models to validate and decipher the roles of these novel genes in disease pathology. The overall goal of our several projects is to identify and characterize novel genes affecting Diabetes and CVD, with a goal of eventual therapeutic development for these diseases. Project 1: Characterization of the role of CYP8B1 in cardiometabolic diseases Using human genetic studies, we have identified a novel gene, CYP8B1, with a potential role in lipid and glucose metabolism. We aim to further characterize the role of this gene utilizing iPS-cell derived hepatocyte and beta-cell models, as well as animal models, in order to derive mechanistic insight into the impact of this gene in cardiometabolic diseases. Project 2: Characterization of the role of ABCA8 in Diabetes Utilizing Mendelian genetic studies in human family cohorts, we have identified ABCA8 as a novel gene linked to both Diabetes and cardiovascular diseases. We aim to decipher the role of ABCA8 in these diseases utilizing human studies, mouse models, and in vitro iPS and cell culture models. Project 3: Characterization of the role of novel genes in lipid and glucose metabolism Using a deep sequencing approach in a human cohort, we have identified genes associated with lipid levels. Genetic studies in humans suggest roles for these genes in lipid/glucose metabolism. We aim to use in vivo CRISPR based approaches to validate the role of these genes in mouse models, as well in ex vivo cell culture models.

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Department of Obstetrics & Gynaecology Principal Investigator

Project Title with a brief description

A/Prof Chan Shiao-Yng [email protected]

Metformin treatment in gestational diabetes and impact on offspring in utero programming

Co- supervisor:

Gestational diabetes mellitus (GDM) complicates 20% of pregnancies in Singapore, one of the highest incidences worldwide. GDM is associated with a range of adverse pregnancy and neonatal outcomes. Importantly, offspring exposed to maternal hyperglycaemia in utero demonstrate increased rates of obesity, diabetes mellitus, adverse metabolic, cardiovascular and renal profiles in childhood and adulthood. Such risks are hypothesised to be transmitted through epigenetic programming of the fetus. Metformin is a biguanide which decreases insulin resistance through several mechanisms and is used in the treatment of GDM in many Western countries, but not routinely in Singapore. Whilst perinatal outcomes with metformin treatment are comparable to treatment with insulin, unlike insulin, metformin crosses the placenta to the fetus. Emerging data suggests that in utero metformin exposure may have a longer-term effect on the offspring in addition to the impact of GDM per se. With metformin exposure in utero, toddlers at age 1 year display a higher body mass index and children at aged 8 years have an increased mean circulating plasma glucose level. Using in vitro models of human embryonic stem cells and human mesenchymal stem cells, this project will investigate the effects of metformin exposure on the growth, development and function of these cells as they differentiate along the adipose and pancreatic cell lineages respectively. Specifically the epigenetic modifications of DNA methylation, histone modifications and miRNA will be assessed. The understanding of such mechanisms will help clinicians re-evaluate the pros and cons of metformin treatment in GDM. The methodologies developed in the project could also underpin the evaluation of future pharmacological interventions in pregnancy.

Dr Walter Stunkel [email protected] Dr Adrian Teo [email protected]

A/Prof Chan Shiao-Yng [email protected]

Placental lipid flux in gestational diabetes as a potential therapeutic target in mitigating the transgenerational cycle of metabolic disease Gestational diabetes mellitus (GDM) is glucose intolerance that is first recognized during pregnancy and it is linked with adverse obstetric outcomes, fetal macrosomia, and future offspring risk of obesity and type 2 diabetes (T2DM). In Singapore, GDM complicates 20-25% of pregnancies and T2DM accounts for over 10% of disease burden in the general population. The transgenerational cycle of diabetes begetting diabetes needs to be stopped in order to effectively fight our war against diabetes. The placenta is central in determining the in utero environment within which the fetus develops. In addition to regulating nutrient-waste exchange between the mother and fetus, it also ensures that fetal requirements are balanced with maternal resources available to support the pregnancy

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using a range of signaling pathways. The placenta plays a large part in transmitting cues to the developing fetus to prepare it for the challenges of the environment it could face following birth by imposing a “programming” effect that is sustained into ex utero life. However, such evolutionary mechanisms could have unintended consequences in pathological conditions like GDM. We hypothesise that a critical biological pathway through which this occurs is the regulation of placental lipid flux. Lipids are not just structural components of cell membranes and sources of energy but lipid species are also biological signals in their own right. Evidence for a significant alteration in the uptake, metabolism and release of lipids during the maternal-fetal lipid transfer across the placenta is starting to emerge in GDM and could be a potential intervention target. In this study, we aim to use a range of ex vivo and in vitro techniques (including human tissue culture, molecular biology, labelled fatty acid uptake, LC-MS lipidomics, pharmacotherapeutics) to better understand the specific dysregulation occurring in lipid flux within placenta from GDM pregnancies and investigate how this could be manipulated by various nutritional supplements and drugs. Results will also be correlated with a range of clinical characteristics measured longitudinally during pregnancy and later in the offspring, and will provide an unprecedented opportunity to investigate and understand the placental mechanisms underlying altered offspring growth and development. The evidence gained from this study could potentially lead to effective prevention and intervention strategies for GDM and mitigating the transgenerational cycle of metabolic disease.

Assistant Professor Citra Mattar [email protected]

Educating Maternal Immunity to improve donor cell engraftment: The kinetics of maternal immune cell trafficking and a novel therapeutic strategy in a murine Monogeneic diseases affect 1:200 fetuses and the most common hereditary condition in Asia and SE Asia are the thalassaemias, with an 8% carrier rate. These disorders cause various pathologies during fetal development and the postnatal period, such as severe haemolytic anaemia, and there are numerous advantages to in utero treatment. Our current research focuses on the safety and efficacy of intrauterine gene and stem cell therapy in small and large animal models for thalassaemia major, as we currently explore both in vivo lentiviral delivery (LV-IUGT) and various combinations of intrauterine haemopoietic cell transplantation (IUHCT) in a thalassaemia major mouse model. Both IUHCT and IUGT offer the potential advantages to effectively cure haemoglobinopathies over postnatal therapy. However, significant hurdles exist in achieving significant functional engraftment or improvement of anaemia. In our IUHCT model haemopoietic stem cells harvested from donor mice are injected intraperitoneally into E14 fetuses of time-mated thalassaemic mice; donor cell engraftment is monitored serially in peripheral blood with terminal harvests to determine organ engraftment. Strategies to improve engraftment include postnatal transplantations with immune-modulation of the recipient to maintain

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peripheral tolerance. In the LV-IUGT model we directly inject a clinical lentiviral vector into murine fetuses at E14 and determine the haematological outcomes serially. We are currently optimizing the dose and delivery of this strategy. Recent data confirm the presence of a formidable barrier formed by transplacentally-trafficked cells and antibodies from the maternal immune system. By understanding the important interaction between maternal and fetal immune systems we will be able to develop strategies to overcome this barrier to generate long-term engraftment that necessary for cure of such genetic diseases. Therefore, another aspect of our research focuses on investigating cell trafficking by identifying different biomarkers that appear on maternal and fetal immune cells maternal. A/Prof Fong Chui Yee [email protected]

Studies on the effects of gestational diabetes mellitus on the fetus using stem cell-derived tissues

Prof Ariff Bongso [email protected]

Gestational diabetes affects about 20-25% of pregnant women and the infants of such diabetic mothers have an increased risk of glucose intolerance, obesity, metabolic syndrome, hypertension and cardiovascular disease. Maternal hyperglycaemia leads to fetal hyperglycaemia because glucose readily traverses the placenta. After 20 weeks of gestation the fetus has a functioning pancreas and is responsible for her own glucose homeostasis because maternal insulin does not cross the placenta in appreciable amounts. Unchecked fetal hyperglycaemia results in hypertrophy of fetal pancreatic islets and hyperinsulinemia. Epidemiological studies show that the consequences of this are life-long and even extend to the next generation through the maternal line. How the mother’s hyperglycemia affects her infant is poorly understood thus hampering strategies for prevention and treatment. Using stem cell technology, we wish to generate pancreatic tissues that would mimic those of infants of diabetic mothers to study the effects of maternal hyperglycemia. We have isolated mesenchymal stem cell populations from the Wharton’s jelly of the human umbilical cord (hWJSC). Besides their painless harvest, these hWJSCs are primitive, abundant, proliferative, safe, hypoimmunogenic and can be differentiated into some desirable tissues. However, attempts at differentiating hWJSCs into true insulin-producing pancreatic tissues have been unsuccessful. We therefore reprogrammed hWJSCs from normal human umbilical cords into induced pluripotent stem cells (iPSCs) using an integration-free, non-viral approach to generate a wider variety of desirable tissues for our research. Our method eliminates the potential risk of accumulation of any virally-induced genetic mutations in the iPSCs. Our reprogramming protocol can now be applied to hWJSCs from the umbilical cords of mothers with pre and gestational diabetes (GDM)

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and the differentiated tissues from these iPSCs provide an unprecedented opportunity to study the effects of maternal hyperglycaemia on the fetus or infant. Our experiments are designed to (i) to reprogram hWJSCs from umbilical cords of GDM mothers into iPSCs, characterize them for stemness and compare their differences with iPSCs derived from hWJSCs of normal cords, (ii) differentiate the GDM-hWJSC-iPSCs into pancreatic insulinproducing cells and compare their phenotype, genotype and function with that of normal-hWJSC-iPSC-insulin-producing cells and (iii) comparatively evaluate the response of GDM and normal hWJSCiPSC-derived insulin-producing cells to constant and pulsatile doses of glucose. The generation of islet-like cells at various stages of maturity from normal and GDM-hWJSC-iPSCs can be used to examine whether the pathogenic mechanisms of GDM are due to elevated sugar levels, fetal hyperinsulinemia or an overworked fetal pancreas. The hWJSC-iPSC-derived insulin-producing cells may also serve as an ideal immunological match for regenerative cell-based therapy for the children of GDM mothers. We hypothesize that stem cells from the Wharton’s jelly (hWJSCs) of the umbilical cord of the mother with gestational diabetes mellitus (GDM) can be reprogrammed to induced pluripotent stem cells (iPSCs) which can be differentiated into insulin-producing cells to study the effects of maternal hyperglycaemia on her fetus. Our specific aims are to (1) reprogram hWJSCs from umbilical cords of mothers with pregestational and gestational diabetes mellitus into iPSCs, characterize them for stemness using the gold standards for pluripotency and compare their differences with iPSCs derived from hWJSCs of normal cords, (2) differentiate the GDM-hWJSC-iPSCs into pancreatic insulinproducing cells and compare their phenotype, genotype and function with that of normal-hWJSC-iPSC-insulin-producing cells and (3) comparatively evaluate the response of GDM and normal hWJSC-iPSC-derived insulin-producing cells to constant and pulsatile doses of glucose. Prof Yong Eu Leong [email protected] Dr Susan LOGAN [email protected]

Updated in Jan 2018

Integrated Women’s Health Program (IWHP) Singapore is globally one of the most rapidly aging societies. With improvements in health care, life expectancy in females has increased from 68 years to 85 years over the past 45 years. In 2013, women aged >45 years comprised 40% of Singapore’s female population &, as such, would expect to spend half of their lives in the menopausal transition & menopause. Many apparently well midlife Singaporean women attend gynecology for routine health screens and care. Currently this care is specialty centric but with a recognition that this age group face multiple health issues including problems relating to menopausal symptoms, bone health and osteoporosis, sexual health, cancer, cardiovascular disease, emotional health, mobility, balance & muscle strength. Many are unrecognized & unmet and all directly influence each other. Improving health care Page 31

delivery and outcomes depends on systematically establishing the prevalence of health conditions and ultimately health-care needs. In addition, previous research has primarily focused on men. Gynaecologists are ideally placed to bring the focus to women & introduce individualized and innovative healthcare solutions for the prevention and timely treatment of chronic conditions such as hip fractures, cardiovascular disease/stroke, dementia, cancer, and emotional and healthy relationships. The IWHP consists of five elements: 1. A cross sectional study to assess the prevalence & correlates of health conditions and risk factors across multiple systems (cardiovascular, mental, musculo-skeletal, sexual and gynaecological) among a clinical population of women ages 45-69 years in Singapore. The assessment tool includes a questionnaire (electronic in 9/2015), biophysical measurements, blood sampling, short physical performance battery & whole body (bone, muscle, fat) DXA. a. To use this cross sectional information to compare the prevalence of health conditions in the IWHP to that of previously published findings from samples of women living in the United States & globally b. To use the fasted blood samples to explore novel biomarkers in relation to disease prediction 2. To integrate research findings within a treatment setting. Participants will be provided results from their research assessments and will be given referrals to internal resources. This model of care will be termed the Integrated Women’s Health Program (IWHP). 3. Using the IWHP as scaffolding for recruitment of eligible patients, it is proposed to initiate interventional clinical trials after identifying specific needs 4. To undertake a longitudinal assessment of this mid-life female cohort Department of Paediatrics Principal Investigator

Project Title with a brief description

A/Prof Lee Yung Seng [email protected]

For MSc or PhD project Prenatal, perinatal, and postnatal determinants of adiposity and metabolic phenotype in childhood This research is part of the birth cohort study “Growing Up in Singapore Towards healthy Outcome” (GUSTO) of the TCR flagship programme “Developmental Origins: Singapore” (DevOS). Prenatal, perinatal, and possibly early postnatal factors determine the subsequent growth of the child. This project aims to uncover the factors which determine the size and body composition of the newborns, and predictors of the growth of young children in the first few years of life, including maternal factors (diet and nutrition, habitus, pregnancy complications), birth events, and the baby’s growth pattern in the first few months’ of life. Specifically the

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candidate will determine the factors which predict catch up growth in the group of children with lower birth weights, as well as those with growth failure. The student will get opportunities to study epigenetic changes of candidate genes which predict the subsequent growth pattern of the children, as well as predictors of failed catch up growth. Determining body composition of infants and young children This research is part of the birth cohort study “Growing Up in Singapore Towards healthy Outcome” (GUSTO) of the TCR flagship programme “Developmental Origins: Singapore” (DevOS). The candidate will study the various modalities of body composition assessment of infants and young children, including MRI of abdominal fat compartments and whole body fat, bioelectrical impedance, and air displacement plethysmography. There will also be opportunity to study correlation of the body composition parameters with developmental factors and epigenetic biomarkers. A/Prof Lee Yung Seng [email protected]

Effects of a BCAA (branched chain amino acids)-supplemented hypocaloric diet versus a standard hypocaloric diet on weight loss, lean mass preservation and improvement of insulin sensitivity, and its benefits over a high-protein hypocaloric diet. In obesity treatment, high-protein diets have shown to be more effective than conventional low-fat, diets in increasing lean mass preservation, but are costly and have potential health risks. Lean mass preservation is crucial for sustaining high basal energy expenditure for greater initial weight loss, mitigating weight regain and improving insulin sensitivity. Branched-chain amino acids (BCAA) supplements are known to stimulate protein synthesis and preserve lean mass but their effects during weight loss have not been investigated. We propose to investigate if a BCAAsupplemented hypocaloric diet when compared to a standard, moderate-protein hypocaloric diet will result in greater lean mass retention and improvement in insulin sensitivity. Additionally, we investigate if the BCAA-supplemented diet has added advantage to a high-protein hypocaloric diet by leading to less detrimental effects on bone mineral density and renal function. Using a 16-week weight loss intervention, combined with 8 weeks of weight maintenance, overweight and obese men and women will be randomized to one of three diets: a hypocaloric diet with BCAA supplements, a standard hypocaloric diet (as control) or a high-protein hypocaloric diet with placebo supplements. Participants’ compliance to the diet versus supplements will be compared. Body composition and bone mineral density (by DEXA), resting and diet-induced energy expenditure (by indirect calorimetry and test meals) and insulin sensitivity (by HOMA) will be measured before and after weight loss. Blood and urine samples will be analyzed for biochemical parameters and renal function. Body composition and bloods will also be measured at the end of weight maintenance. The effect of the diet intervention will be assessed by using repeated-measures ANOVA. Findings of this

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study will provide novel insight on the use of BCAA-supplementation during energy restriction and may offer an alternative cost-effective strategy for weight loss and maintenance, without the adverse health effects of a high-protein load. Whole-exome sequencing to identify genetic variants associated with severe childhood obesity, and tracking the changing prevalence of obesity related complications Susceptibility variants contributing to obesity are being uncovered by candidate gene approach, genome wide linkage studies, and genome wide association studies (GWAS), but the collective effect is small and does not explain most of the BMI heritability. We propose a whole exome sequencing (WES) approach to systematically interrogate coding variants and to identify novel obesity loci in 130 Chinese subjects from our existing cohort of Singaporean severe early-onset obese subjects, and an additional ~200 subjects will be recruited to augment this. Sequencing will be performed at high coverage (50X) on the Illumina-HiSeq platform. Statistical association tests will be performed for identified variants (single-variant analysis and gene-based mutational load test) using normal, lean (BMI between 18.5 and 23 kg/m2) adult Singaporean Chinese individuals as controls (N=500). A stringent approach will be undertaken to identify and prioritize novel obesity variants, including checks with genotype data, through bioinformatics tools for predicting functional effects and by re-sequencing and/or de novo genotyping of top hits. We will replicate top hits by targeted sequencing in 96 (out of 200 subjects) extreme obese Chinese children and through de novo genotyping in additional Chinese children with extreme BMI from childhood datasets from Singapore and China. We will study the invitro functions of the aberrant proteins of novel variants. Our secondary aim is to assess the change in prevalence of obesity related complications over 15 years by recruiting 350 severely obese children and compare against the complications prevalence data obtained from an NMRC funded childhood obesity study in 20012004. The identification of novel genes for extreme early-onset obesity can provide novel insights into human weight regulation mechanisms and new therapeutic targets for drug development, and also identification of high-risk children so that potential intervention measures can be implemented at an early stage of life. A/Prof Lee Yung Seng [email protected]

For PhD project Diet during infancy ( 0 -5 years) and infant health outcomes

Co- supervisor: Dr Mary Chong [email protected]

Updated in Jan 2018

The early years of life are critical to the growth, development, and well-being of children. Feeding practices and food consumption contribute to infants' nutrient adequacy or excess and also establish dietary patterns and habits that can last a lifetime. Understanding food consumption patterns and nutrient intake is important to identify areas for improvement in establishing healthy eating patterns at an early age. Page 34

Compared to their Western counterparts, little is known about the food consumption patterns and nutrient intakes of Asian infants. The existence of the three major ethnic groups (Chinese, Malay and Indians) in the local Singapore population provides an ideal backdrop for a study to investigate the dietary patterns and nutrient intakes of infants in the Asian region. This research will use data from the GUSTO (Growing Up in Singapore Towards healthy Outcomes) mother-offspring cohort study in Singapore and the largest in Asia. It is a longitudinal study designed to understand the developmental origins of health and disease from an Asian perspective. A cohort of 1200 pregnant women were recruited during the first trimester of pregnancy and followed up regularly throughout pregnancy and the nutrition, health and development of the child tracked from birth to 8 years of age. Research aims: With this, research projects would be undertaken to examine the following hypotheses: 1) What are the dietary patterns of Asian children in the early years of life ? Are there differences between ethnic groups? 2) What are the associations between nutrient intakes, dietary patterns of Asian children and their growth and adiposity in the early years of life ? 3) Does sleep duration in early life influences energy intakes and dietary patterns in Asian children and modifies the effect on growth and adiposity? A/Prof Lee Yung Seng [email protected]

For MSc project Role of sleep in behavioral and metabolic outcomes

Co- supervisor: A/Prof Joshua J. Gooley [email protected]

Updated in Jan 2018

Good sleep is important for good health. Over the past decade, increasing evidence suggests that short sleep negatively impacts cognition and contributes to increased risk of weight gain and metabolic disorders. As part of the Growing Up in Singapore Towards healthy Outcomes (GUSTO) birth cohort study in Singapore, we are investigating the influence of sleep behavior on neurobehavioral and growth outcomes. We are examining sleep behavior in children and their mothers using questionnaires and actigraphy. Our aim is to characterize the role of sleep in mental and physical health, as well as factors that influence sleep duration and sleep quality.

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Department of Obstetrics & Gynaecology Principal Investigator

Project Title with a brief description

A/Prof Chong Yap Seng [email protected]

For PhD project

Co- supervisor: Dr Walter Stunkel [email protected]

The role of epigenetic pathways present in umbilical cord derived mesenchymal stem cells in fetal programming of metabolic disease It has been shown that an adverse intrauterine environment can largely influence the risk for non-communicable diseases later in life. Maternal gestational diabetes (GDM) and intrauterine growth restriction are correlated with increased adolescent type 2 diabetes, obesity and cardiovascular disease in the offspring. As birth weight per se does not fully explain obesity in the offspring of diabetic mothers, we speculate that epigenetic transgenerational programming is a leading cause for the increased metabolic disease risk susceptibility in these children. The project involves work with established primary cell lines from umbilical cord (Mesenchymal Stem Cells, MSCs and Human Umbilical Vein Endothelial Cells, HUVECs). The cells were taken from growth restricted neonates, as well as those with maternal GDM background and matched with controls. The student will apply state of the art laboratory technologies for assessing the transcriptome, as well as the epigenetic state of MSCs by analyzing genome wide DNA-methylation and histone modifications. MSC lines will be differentiated into various cell types such as adipocytes, myocytes and beta-like cells. The goal of the project will be to functionally characterize candidate genes which may play an important role in the development of metabolic diseases. These efforts may lead to novel biomarker hypotheses for patient stratification, as well as potentially supporting the identification and validation of new therapeutic targets.

Department of Physiology Principal Investigator

Project Title with a brief description

Prof Carlos Ibanez [email protected]

Growth Factor Receptor Signaling and Biology in Nervous System Development and Metabolic Regulation With a focus on growth factor receptor signalling, the aim of this research is to discover novel, general biological principles and mechanisms important for nervous system development and metabolic regulation. We plan to concentrate the main thrust of our future work on two programmes that complement and expand current areas of expertise. The first one focuses on the mechanisms of action and biological activities of the death receptor p75NTR (p75 neurotrophin

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receptor), and reflects our continued interest in neuronal growth factors, their signalling mechanisms and their potential for the development of novel therapies for neurodegenerative diseases and neurotrauma. This is a program that involves signal transduction studies, mouse genetics approaches, and a drug discovery effort to identify novel p75NTR inhibitors. The second program focuses on the role of a novel regulatory network, formed by members of the transforming growth factor-beta (TGF-) s upe rfa m ily a nd the ir re ce ptors , in the control of glucos e homeostasis, fat accumulation and energy balance. It is based on our original discovery of the TGF- s upe rfa m ily re ce ptor ALK7 a nd our recent studies on the functions of activin signalling through ALK7 and ALK4 in pancreatic islets, adipose tissue, and hypothalamus. This program offers a new inroad to the study of metabolic regulation and involves complementary studies in cell culture as well as conditional and knock-in mouse models. Dr Karthik babu Mallilankaraman Genome-wide RNAi screening to discover novel modulators of endothelial cell death and their molecular mechanism of action. [email protected] Reperfusion of the ischemic tissue is a major trigger and contributor of cell death in a variety of cardiovascular, metabolic and neurological diseases. Cells and tissues exposed to hypoxia during ischemia may sustain damage and cell death upon reoxygenation by restitution of blood flow. This Hypoxia/reoxygeneation (H/R)-related injury has long been associated with the enhanced production of reactive oxygen intermediates generated during reoxygenation. Additionally, H/R may cause injury through the recruitment of proinflammatory leukocytes. Endothelial cells represent the first cell type injured by reactive oxygen species generated during H/R as these cells form the first line of defense in the vasculature. In vitro, the mechanisms of endothelial cell death caused by H/R remain poorly understood and may involve both necrotic and apoptotic forms of cell death. Recent advances in the genomics have rendered researchers to develop various tools to dissect molecular pathways in complex diseases. Specifically, the discovery of RNA interference (RNAi) and the rapid development of technologies exploiting its functional applications have enabled us to examine the consequences of lossof-function of a particular gene product in mammalian cells. Completion of human genome project further advanced the field towards developing genome-wide RNAi libraries targeting the entire human genome. Genome-wide RNAi libraries have made it possible to carry out large scale, phenotype-based screens, which have yielded potent information on diverse cellular processes ranging from oncogenic to infectious diseases. Nevertheless, such screening methodologies have never been employed in vascular diseases. We have developed an unbiased genome-wide screening approach to identify novel modulators of cell death. My lab will use Updated in Jan 2018

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this approach to identify previously unknown molecular pathways that exist in endothelial cells during Ischemia/Reperfusion injury. Category: Infectious Disease Department of Biochemistry Principal Investigator

Project Title with a brief description

A/Prof Gan Yunn Hwen [email protected]

The role of glutathione in modulating host immune response as well as bacterial virulence. The project will examine how glutathione, an essential molecule that regulates host redox state affect the transcription of host immune response genes and how it modulates bacterial behaviour inside host. There will be an emphasis on enteric bacteria and the possible role this have in affecting our gut microbiota. Previous publications from my lab that are directly relevant to this project: Tan et al, 2012, J Clin Invest 122:2289. Wong et al 2015 Cell Host &Microbe 18:38.

A/Prof Tan May Chin Theresa [email protected]

Role of microRNAs in Hepatitis B virus replication and reactivation MicroRNAs (miRNAs) are small RNA molecules of ~21-25 nucleotides which function as important regulators of gene expression. Studies have shown that miRNAs can influence diverse pathways and affect many physiological processes including metabolism, development, differentiation and apoptosis. MiRNAs have also been shown to be involved in many diseases including cancer. Of relevance to this study is the observation that miRNAs can impact viral infection and replication. In addition, miR-122 is the first miRNA being used as a therapeutic target in humans. MiR-122 is critical for the replication of hepatitis C virus (HCV) and it binds to 2 sites on the 5’ non-coding region of the HCV RNA. Phase 2a trials of miravirsen, a locked nucleic acid modified antisense oligonucleotide complementary to part of miR-122, have resulted in significant reduction in viral RNA with no emergence of resistant viruses. MiRNAs have been shown to affect hepatitis B virus (HBV) gene expression and inhibit HBV replication. MiRNA-199a-3p. miRNA-210 and miR-122 were able to suppress hepatitis B virus replication by directly binding to the target regions of HBsAg transcripts, or interfere with the transcription factors that regulate HBV replication. Members of the oncogenic miR-17-92 cluster have also been shown to affect HBV replication. miR-20a and miR-92a-1 can directly bind to HBV genes and inhibition of these miRNAs enhances HBV replication. However the comprehensive studies of miRNAs which are critical for HBV replication and reactivation are still lacking.

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Given the above observations, the specific aims for this study are: To identify miRNAs which are critical for HBV replication. To identify miRNAs which are crucial for hepatitis B reactivation in patients. To test the possible use of miRNAs as therapeutic targets for inhibition of HBV replication using the humanized mouse model. A/Prof Tang Bor Luen [email protected]

Exploring the role of Rab35 in bacterial infection and bacteriacontaining vacuoles (BCVs) Intracellular bacterial pathogens drive the formation of host membrane-derived pseudo-organelles that facilitate their replication, survival, or dormancy. The formation and maintenance of these BCVs are dependent on the bacteria's ability to usurp the host's intracellular membrane system, in particular dynamic compartments involved in exo-/endocytic membrane traffic and autophagy. Bacteria are typically internalized by phagocytosis, and the compartment matures through endosomal fusion. The bacteria-containing phagosome/endosome often becomes the base for BCV formation. Diverse strategies used by different bacterial pathogens prevent the BCV from being destroyed via the endolysosomal pathway. Furthermore, bacterial survival or proliferation in BCVs could be augmented by host membrane transport processes subverted by secreted bacterial factors, which facilitate the acquisition of membrane sources and nutrients. Rab35, a small GTPase of the Rab family, are known to modulate endosomal transport processes in a multitude of physiological and pathological conditions, including cell division and cancer. We have recently shown that Rab35 facilitates uropathogenic E. coli (UPEC) survival in UPEC-containing vacuoles (UCV) within infected bladder cells in vitro and in vivo. The bacteria enhance the expression of both Rab35 and TfR and recruit these proteins to the UCV, thereby supplying UPEC with the essential nutrient iron. Much remains to be explored with regards to the general cellular mechanisms of Rab35 function with regards to BCV formation and maintenance, and intracellular survival of bacterial invaders. We shall investigate cellular mechanisms and pathways modulated by Rab35 that are physiological, as well as bacteria-influenced, with a range of biochemical, cell biological and imaging techniques. The proposed research project shall be done in collaboration with Dr Bindu Sukumaran of Duke-NUS. References and readings: 1. Tang BL (2015) Bacteria containing vacuoles – subversion of cellular membrane traffic and autophagy. Crit. Rev. Eukaryot. Gene Expr. 25: 163-174. 2. Dikshit N, Bist P, Fenlon SN, Pulloor NK, Chua CE, Scidmore, Carlyon JA, Tang BL, Chen SL and Sukumaran B. (2015) Intracellular uropathogenic E. coli exploits host Rab35 for iron acquisition and survival within urinary bladder cells. PLoS Pathog. 11(8): e1005083.

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A/Prof Yew Wen Shan [email protected]

Current Synthetic Enzymology research interest focuses on protein engineering and biocatalysis, with emphasis on using structural and mechanistic enzymological knowledge to develop therapeutics. We are involved in the following research areas: 1. Defining Synthetic Enzymology as an enabling platform for the

2.

3. 4. 5. 6.

Construction of Next-Generation Synthetic Biology solutions for Pharma and Consumer Businesses. Using Synthetic Enzymology to discover and develop novel therapeutic biomolecules, in the areas of infectious diseases, metabolic disorders, cancer and aging. Developing anti-virulence Quorum-Based Technology for use in the biomedical industry. Using Synthetic Lixiviant Enzymology for Biomining of Electronic Wastes for Environmental Sustainability. Development of Lead Compounds Targeting Enzymes involved in Metabolic Cancer. Using Synthetic Enzymology for the Construction of Therapeutic (Probiotic) Cells for the Treatment of Metabolic Diseases and Infectious Diseases.

Department of Medicine Principal Investigator

Project Title with a brief description

Prof Lim Seng Gee [email protected]

TCR Flagship Programme - THE ERADICATION OF CHRONIC HEPATITIS B

Dr Lee Guan Huei [email protected]

The Singapore Hepatitis B Consortium (SHBC) is a national translational group of 30 researchers and scientists who have been working collaboratively and now plan to focus on the eradication of Chronic Hepatitis B (CHB), through the strategies of examining host and viral interactions, target discovery and validation, and experimental therapeutics, representing three integrated themes. Theme 1: Death Or Survival In Hbv Virus-Host Interactions 1.1 Identify Human-host response to hepatitis B viral infection: 1.1.1 Identify Human Factors that lead to viral eradication 1.2 Identify Viral factors that influence disease outcome 1.2.1 Viral factors that influence disease outcome 1.3 Identifying underlying immune mechanisms regulating disease outcome 1.3.1 Examine Intrahepatic Immune Responses to HBV 1.3.2 Examine the Emerging role of innate immunity to HBV 1.3.3 Evaluate The role of T cell regulation in CHB 1.3.4 Identifying and characterising the “lost” HBV specific T cells Theme 2: Target Discovery and Validation 2.1 To identify new key host nuclear factors interacting with HBV

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covalently closed circular DNA (cccDNA) 2.2 To identify miRNAs critical for HBV replication and reactivation 2.3 To Identify HBV genotype specific changes in gene expression in the HBV-infected cells and HBV molecular regulation by endogenous host proteins 2.4 To develop Clustered Regulatory Interspaced Short Palindromic Repeats (CRISPR) associated (Cas)9-based RNA-guided nucleases (CRISPR/Cas) targeting HBV-specific sequence as a novel strategy to disrupt cccDNA in hepatoma cell line and animal models leading to viral eradication 2.5 To develop a novel high-throughput/ high content therapeutic screening platform for anti-HBV drug development using a variety of focused small molecule compound libraries T12.3: Theme 3: Experimental Therapeutics 3.1 HBV Experimental Therapeutics in a humanised mouse model 3.1.1 Optimisation of Humanised Mouse Model for HBV infection 3.1.2 Creation of phenotype specific CHB as models of personalised medicine 3.1.3 Therapeutic manipulation of CHB humanised model to optimise HBV eradication 3.2 Clinical Experimental Therapeutics 3.2.1 The impact of viral antigen load reduction on human immune cells by plasmapheresis in high viral load patients with chronic hepatitis B 3.2.2 Examine HBsAg Seroclearance in a Randomised Control Trial of peg-interferon of Chronic Hepatitis B patients with low viral load versus no treatment (INACTIVE study) 3.2.3 Examine HBsAg Seroclearance in a Randomised Control Study of Nucleos(t)ide Treated Patients Randomised to Tenofovir, or Tenofovir + Telbivudine (DUAL Study) For more details, please see https://drive.google.com/file/d/0BzM8YhxbhxfMjhBd05nQlN3WEk/view?usp=sharing Prof Nicholas Paton [email protected]

Development of New Treatments for Tuberculosis Tuberculosis (TB) is one of the world's most threatening infectious diseases with an estimated 8.6 million new cases and 1.3 million deaths per year. It is caused by Mycobacterium tuberculosis, which is becoming increasingly resistant to current drugs, thereby posing a major public health problem in Asia and worldwide. Our group’s research is focused on creating improved, more efficient treatments for TB. We are starting several projects aimed at investigating new promising TB drug candidates that need to undergo thorough preclinical evaluation (including animal efficacy studies) before proceeding to clinical development. The project activities will range from biochemical and whole cell assays, mechanism of action

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studies and mouse model testing. The latter will include novel stateof-the-art imaging approaches such as PET/CT, which will enable to monitor TB relapses longitudinally. Given strong supporting results in the pre-clinical studies, the candidate compounds will proceed to clinical studies for safety evaluation and whole blood bactericidal activity assays (WBA). WBA is an ex vivo model for assessing mycobacterial sterilization by drugs administered to human subjects. Due to the project specifics and compulsory biosafety requirements, we seek PhD candidates who: - have at least 1 year of documented BSL-2 lab experience (compulsory), - possess basic knowledge of molecular biology, - show strong interest in infectious diseases and microbiology, - are willing to work in a BSL-3/ABSL-3 environment, - prior experience with mycobacteria, work with laboratory animals, and/or BSL-3 is highly desirable. The PhD candidate in this project will have a unique opportunity to gain in-depth expertise in translational bench-to-bedside drug development, will acquire specialized in-demand work skills in a BSL3 laboratory environment, learn the principles of clinical studies, be part of a multidisciplinary project team and transform data into knowledge for improving human lives. Department of Microbiology and Immunology Principal Investigator

Project Title with a brief description

A/Prof Sylvie ALONSO [email protected]

Project 1 - Testing the in vitro and in vivo efficacy of dengue therapeutic candidates In this project, the protective efficacy of selected therapeutic candidates will be tested in vitro and in vivo using various mouse models of dengue. The work involves virology basic techniques, cell biology and extensive mouse handling. Project 2 - Characterization of the antibacterial and antiinflammatory properties of Q203, a promising clinical candidate for the treatment of multi-drug resistant tuberculosis. A previous study reported that Q203, a clinical drug candidate for the treatment of Tuberculosis, completely inhibits the development of granulomatous inflammation in the lung of infected animals. This is a very interesting observation that seems to be specific to Q203 since other TB drugs such as isoniazid or bedaquiline do not inhibit granuloma formation. It is however unknown whether the inhibition of granuloma formation is a pre-requisite for the mycobactericidal activity of Q203 in vivo. In this project, we thus propose to address this hypothesis by characterizing the anti-inflammatory properties of Q203 under various experimental conditions. In vivo (mouse) and in vitro (using mammalian cell lines) experiments

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will be performed. Project 3 - Survival strategies of pathogenic mycobacteria within its host macrophage In this project, the aim is to study the role of selected gene candidates in the survival of pathogenic mycobacteria (BCG and M. tuberculosis) within macrophages. The techniques include molecular biology, microbiology, cell biology and possibly mouse handling. Dr CHEN John Yu-Shen [email protected]

Phage interference of staphylococcal pathogenicity islands counteracts molecular parasitism The Staphylococcus aureus pathogenicity islands (SaPIs) are prototypical members of a family of highly mobile phage-inducible mobile genetic elements that encode virulence factors and are parasitic on certain temperate (helper) phages. Normally integrated at specific chromosomal sites, the SaPIs remain quiescent until helper phage-encoded anti-repressor proteins lift their repression. Once the SaPI life cycle is set in motion, SaPI-coded mechanisms that interfere with the life cycle of the phage maximize SaPI DNA encapsidation into infectious phage-like particles. This project is based on the concept of a “molecular arms race” between the SaPIs and staphylococcal phages, and the aim will be to identify and characterize phage-coded mechanisms that interfere with the SaPI life cycle. Mapping phage-mediated horizontal gene transfer Staphylococcus aureus is naturally susceptible to nearly every antibiotic that has ever been developed; yet, not surprisingly, resistant S. aureus strains have been reported within 1-2 years of the introduction of nearly every antibiotic that has ever been developed. Just as these resistance mechanisms have been evolving, so have the virulence genes been evolving in parallel, and analyses of the growing number of sequenced S. aureus strains have revealed dramatic differences in the carriage of accessory genes, particularly of those involved in pathogenesis. While rapid adaptation is most often the result of horizontal gene transfer (HGT), S. aureus lacks DNA transformation and conjugative plasmids are exceptionally rare, leaving only bacteriophage transduction as the only viable mode of HGT. Phage-mediated horizontal gene transfer is one of the primary driving forces of bacterial evolution. The goal in this project is to begin to dissect the S. aureus metagenome transferred and exchanged by phages (phageome), one phage at a time, to determine if genes important for pathogenicity are preferentially exchanged by transduction.

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A/Prof Vincent Chow [email protected]

Activation and formation of neutrophil extracellular traps in severe influenza and secondary pneumococcal pneumonia Patients infected with highly pathogenic influenza viruses (e.g. H5N1) develop signs of acute lung injury (ALI) with pulmonary infiltration and edema, which further progress into acute respiratory distress syndrome (ARDS) and respiratory failure. Mice lacking IL17RA (necessary for neutrophil recruitment) exhibit decreased morbidity and mortality compared to wild-type mice following influenza virus challenge. Neutrophils play key roles in acute lung injury by producing reactive oxygen intermediates, inflammatory cytokines and enzymes such as elastase, or via formation of neutrophil extracellular traps (NETs), which can cause alveolar destruction and pulmonary edema. Although highly pathogenic viral and bacterial infections cause extensive accumulation of neutrophils in the lungs, how neutrophils mediate the progression of clinical manifestations leading to acute lung injury is not well understood. This project aims to explore the molecular mechanisms by which neutrophils contribute to ALI and ARDS in mouse models infected with influenza A H1N1 and H3N2 viruses, as well as in secondary pneumococcal pneumonia. The objectives include: (1) To evaluate the progressive changes in the lungs and contribution of neutrophils and their mediators in pulmonary complications of ALI/ARDS during primary and secondary pneumonia. (2) To explore opportunities for intervention by treatment with inhibitors against neutrophil recruitment (e.g. P38 MAPK inhibitor), neutrophil elastase and NADPH oxidase which will be tested in combination with antimicrobial agents – this will evaluate whether such interventions alleviate complications of pulmonary edema and alveolar injury. (3) To compare and characterize NETs formation in response to infection with different strains of Streptococcus pneumoniae of varying virulence, and whether antimicrobial combination therapy can ameliorate their deleterious effects. (4) To determine the contributions of pneumococcal virulence factors to NETs formation.

Dr Justin Jang Hann CHU [email protected]

Drug discovery and target validation for medically important mosquito-borne viruses. The re-emerging infections caused by mosquito-borne viruses such as dengue and chikungunya are on the raise at an explosively rate in many parts of the world. In the view of the absence of an effective vaccine and antiviral drug against these viral pathogens, the developments of effective antiviral strategies against these viruses are in high demand. The intent of this study is to perform an established high content cell-based screening platform that could identify compounds that prevent virus replication by (i) inducing antiviral gene expression pathways in host cells, (ii) interfering with viral proteins and their functions (iii) interfering essential host-viral protein interactions. Although the assay can identify compounds that target distinct steps of the virus replication cycle, we are interested in those that stimulate cellular responses necessary to confer initial resistance to virus infection. Bioactive compound libraries will be

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screened on the high content cell-based assay in triplicate. All of the compounds that meet the designated criteria of a “hit” in the primary high content screening platform will be further confirmed with secondary assays (infectious plaque assays, protein over-expression and RNA interference studies). The efficacy of the selected compounds will be further validated in murine models. In additional, the antiviral targets or mechanisms will be dissected by a combination of reverse genetics, proteomics and bioimaging approaches. The research data generated from this study are essential for the rapid identification of potential prophylactic or therapeutic leads that will be helpful in better preventive management of individuals from suffering viral infections. In this proposal, re-emerging flavivirus dengue and chikungunya viruses will be used as the model for the “proof of concept” development. Clinical biomarkers from whole salivary profiling of Hand, Foot and Mouth Disease (HFMD) patients. Hand, foot and mouth disease (HFMD) is a highly contagious illness seen most commonly in young children and immune compromised adults. In the last 15 years, the Asia-Pacific region has seen an increase in both frequency and severity of HFMD outbreaks, several of which reached epidemic levels. Despite the general mildness of the disease, HFMD has risen in status to become one of the top concerns with regards of public health due to the highly contagious nature of its etiological agents, which has placed a strain on healthcare facilities and social-economical stress of the society in the region during major outbreaks. The disease can be caused by multiple serotypes of enteroviruses, most commonly members of the Enterovirus A genus, Enterovirus 71 (EV71) and Coxsackievirus A16 (CA16). Currently, there is no specific therapy or vaccine available for HFMD and diagnosis of HFMD is conducted principally through clinical examination. Atypical presentations of HFMD have been associated with several serotypes, including EV71 and such cases may escape clinical diagnosis. The problem is compounded by the higher risk of EV71-infected patients to develop acute neurological conditions rapidly, thereby compromising the prognosis of the affected individuals. Hence, the primary objective of this study is to identify specific biomarkers for HFMD-related enteroviral infections, using high resolution mass spectrometry-based quantitative proteomics technology and miRNA profiling with saliva samples from HFMD patients. Most importantly, the novel clinical and scientific knowledge gained from this study will definitely accelerate the path towards discovery of functional biomarker(s) for rapid diagnosis and detecting the viral disease progression hence allows for better clinical management of patients and the formulation of intervention plans for this emerging infectious disease.

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A/Prof Tan Yee-Joo [email protected]

Project title 1: Studying viral-host interactions in hepatitis viral infection Description: Approximately 2 billion people have been infected by two viruses, hepatitis B virus (HBV) and hepatitis C virus (HCV), which primarily attack the liver. While some of the infected people clear the virus, a high percentage of them becomes chronically infected. Subsequently, the majority of them develop severe liver disease which includes fibrosis, cirrhosis and eventually hepatocellular carcinoma (HCC). While it is known that infection by HCV/HBV contributes directly and indirectly to the development of HCC in patients, the exact manner by which each virus participates in this complex process is not completely understood. Although HCV and HBV are completely different viruses, both of them preferentially infect hepatocytes and are able to subvert the innate and adaptive immunity, leading to chronic infection. During this process, the complex interplay between viral proteins and host cell machineries contributes to viral replication and/or pathogenesis. Identification of such crucial viral-host interaction therefore offers opportunities for designing new treatments for hepatitis virus-related liver diseases. In this project, we will use a multidisciplinary approach to identify host factors that are regulated by hepatitis viral infection and/or interact with the HCV/HBV viral proteins. The functional significance of novel viral-host interactions identified will be analyzed by using cell culture as well as mouse models of infection. References: 1. ≠Keng, C.T., ≠Sze, C.W., Zheng, D., Zheng, Z., Yong, K.S.M., Tan, S.Q., Ong, J.J.Y., Tan, S.Y., Loh, E., Upadya, M.H., Kuick, C.H., Hotta, H., Lim, S.G., Tan, T.C., Chang, K.T.E., Hong, W., Chen, J., Tan, Y.J.*, Chen, Q* (2015). Characterization of Liver Pathogenesis, Human Immune Responses and Drug Testing in a Humanized Mouse Model of Hepatitis C Virus Infection. Gut, 2015 Jul 6. pii: gutjnl-2014-307856. doi: 10.1136/gutjnl-2014-307856. [Epub ahead of print] (≠equal contributing authors; *co-corresponding authors). 2. ≠Aweya, J.J,, ≠Sze, C.W., Bayega, A., Mohd-Ismail, N.K., Deng, L., Hotta, H., Tan, Y.-J. (2015) NS5B induces up-regulation of the BH3only protein, BIK, essential for the hepatitis C virus RNA replication and viral release. Virology. 474:41-51. (≠equal contributing authors) Project title 2: Generating antiviral neutralizing antibodies and characterizing their mechanism of inhibition Description: Seasonal influenza A virus causes significant morbidity and mortality yearly while newly emerged strains continue to pose pandemic threats. Current strategies against influenza include vaccination and antiviral drug treatment. However, predicting the major strain that may cause the next pandemic is the main obstacle in current vaccine development. Moreover, some viruses have acquired resistance to approved antiviral drugs. Passive immunotherapy is now increasingly being used to treat human infectious diseases and there

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is a demand for the development of neutralizing mAbs for passive immunotherapy in the event of a highly pathogenic flu pandemic as this could be particularly useful for protecting certain groups of people, such as immuno-compromised patients or the elderly, who may not respond well to vaccines. As the hemagglutinin (HA) protein mediates viral entry, it has been the main target for the preclinical studies on antibody-based immunotherapy and these studies suggest that it may be a viable option to administer neutralizing HA mAbs as a form of passive immunotherapy for influenza A infection. However, viral escape mutants were observed when anti-HA mAbs were used individually. Combination therapy, where multiple steps in the virus life cycle are inhibited simultaneously, is highly recommended to minimize the development of escape viruses. Hence, this study uses a multi-disciplinary approach to determine if other viral proteins of influenza A virus can stimulate neutralizing antibodies that prevent viral infection and/or replication. If we can develop combination therapy by using a mixture of mAbs that bind to different components of the virus, this will minimize the chance of drug resistant virus developing and this will help in the establishment of a novel class of antiviral therapeutic drugs. References: 1. Barnwal, B., Mok, C.K., Wu, J., Diwakar, M.K., Gupta, G., Zeng, Q., Chow, V.T., Song, J., Yuan, Y.A., Tan, Y.-J. (2015) A monoclonal antibody binds to threonine 49 in the non-structural 1 protein of influenza A virus and interferes with its ability to modulate viral replication. Antiviral Res. 116:55-61. 2. Mak, T.M., Hanson, B.J., Tan, Y.J. (2014) Chimerization and characterization of a monoclonal antibody with potent neutralizing activity across multiple influenza A H5N1 clades. Antiviral Res. 107:76-83. Dr (Chris) Sham Lok To [email protected]

Elucidating capsular polysaccharide biogenesis in Streptococcus pneumoniae Streptococcus pneumoniae is an important respiratory pathogen that causes more than two million deaths worldwide annually. Similar to other pathogenic bacteria, S. pneumoniae encases its cell envelope with capsular polysaccharide (CPS) which protects itself from opsonophagocytosis and mucus clearance. Because of its importance in pathogenesis, all clinically relevant vaccines against S. pneumoniae target the CPS. There are over 98 serotypes identified in S. pneumoniae. The structural variance in CPS produced is attributed to the extreme diversity of glycosyltransferases (GTs), flippases, and polymerases located in the cps locus. Although these enzymes were identified decades ago, little is known about the molecular mechanisms governing their specificity, regulation, and function. Therefore, the long-term goal of this project is to address this knowledge gap by investigating how specificity is determined by the CPS enzymes. This proposal is based on the wealth of structural information available for pneumococcal CPS, genetic approaches enabled by the conditional essentiality of the CPS pathway, and strong unpublished data on related pathways in E. coli. We will elucidate how

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specificity is determined by the CPS flippases and GTs as well as define the essential residues required for their function. In addition, small molecules that inhibit different steps of the CPS pathway will be identified. Understanding how CPSs are synthesized in S. pneumoniae is critical to improve our current antimicrobial and vaccine strategies. Results from this study will also have a broad impact on our understanding of related polysaccharide pathways such as O-antigen biosynthesis, O-linked and N-linked glycosylation, and exopolysaccharides synthesis. Finally, enzyme variants isolated from this work with relaxed or altered substrate specificity will provide valuable tools for synthetic biology and glycoengineering. Dr Benoit MALLERET [email protected]

Characterization of parasite-proteins involved in Plasmodium ovale and P. malariae invasion Malaria is a mosquito-borne infectious disease affecting 216 million of people per year. Plasmodium falciparum and P. vivax are the main species but due to mass-drug treatment efficiency and the Malaria eradication program coordinated by World Health Organization, P. ovale and P. malariae are more and more prevalent and the resurgence of this neglected species is a recent public health concern in tropical areas. Invasion of red blood cells by the malaria merozoite is an essential step in the life cycle of this obligate intracellular pathogen. A protein family expressed at the apical pole of the P. vivax merozoite, named reticulocyte binding proteins (PvRBP) has been identified and was proposed to have a role in reticulocyte recognition/selection. These gene family is present also in the genome of P. ovale and P. malariae. One of the major hurdle to vaccine development and the discovery of new receptors has been the lack of a reliable assay for assessing the inhibition of invasion induced by antibodies produced in response to various vaccine formulations. We have developed ex vivo methodologies for studying cell biology of P. vivax and P. cynomolgi. These different methodologies are applicable to the other Plasmodium spp. This proposal aims to identify the molecular entities required for P. ovale and P. malariae invasion.

Department of Physiology Principal Investigator

Project Title with a brief description

A/Prof Lina Lim [email protected]

Role of Annexin-A1 in influenza virus infection Annexin-A1: a host factor modulating influenza virus replication The influenza virus infects millions of people each year and can result in severe or even fatal complications. Understanding host responses to influenza infection will enable the development of more effective antiviral therapies. The host immune system recognizes viral RNA via specific receptors and intracellular sensors that directly activate antiviral immune responses. Previous research has revealed diverse yet

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important roles for Annexin family proteins in modulating the course of influenza infection. However, the role of Annexin-A1 (ANXA1) in influenza virus infection has not been addressed. ANXA1 is increased in nasal swab samples obtained from influenza infected patients. In addition, presence of ANXA1 increases virus replication and results in more weight loss after virus infection. We therefore hypothesize that ANXA1 may play a critical role in host anti-viral responses. To test this hypothesis, we propose to assess how ANXA1 expression and activation in different cell types can influence the immune response to influenza in vivo and to determine the molecular role played by ANXA1 in the innate immune response to influenza. Category: Neuroscience Department of Anatomy Principal Investigator

Project Title with a brief description

A/Prof Ong Wei Yi [email protected]

Role of Sphingomyelinases in Neurological Disorders Sphingomyelinases, which catalyze the hydrolysis of sphingomyelin to ceramide and phosphorylcholine, are abundant in the brain. These enzymes are a major, rapid source of ceramide production not only during physiological responses to receptor stimulation, but also in neurological disorders. The project will start by introducing students to sphingomyelinases and its enzymatic product ceramide, in membrane domains or lipid rafts and the nucleus. This is followed by practical work on determination of crosstalk between sphingomyelinase and cytosolic phospholipase A2 (cPLA2) catalysed products including arachidonic acid, functions of acid sphingomyelinase (aSMase) and neutral sphingomyelinase (NSMase) in neurons, neuronal progenitor cells, glial cells, and brain endothelial cells; alterations in acid and N-SMases in Niemann Pick Disease Type A, major depression, Alzheimer's disease, cerebral ischemia, and pain If time permits, novel strategies sphingomyelinases will be explored.

A/Prof Samuel Tay [email protected]

to

explore

inhibitors

to

The role of microRNAs in the pathogenesis of Parkinson’s disease in vivo. MicroRNAs are non-coding RNAs which have been implicated in several neurodegenerative diseases. Our group have shown that mi124 is directly involved in the calpain/CDK pathway, which

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ultimately lead to the death of dopaminergic neurons in the substantia nigra in the mid brain of MPTP-treated mouse model of Parkinson’s disease. We hope to extend our research work to cell culture system using cell-lines to understand the functions of these miRNAs in the generation of molecular events that leads to dopaminergic neural cell death and the pathogenesis of Parkinson’s disease. Department of Biochemistry Principal Investigator

Project Title with a brief description

A/Prof Tang Bor Luen [email protected]

Rab31’s roles in regulating astroglia differentiation and function A key family of molecules regulating the flow of membrane cargoes in eukaryotic cells is the Rab family of small GTPases. The largest subfamily of the Ras superfamily of small GTPases, there are more than 60 members of the Rab family genes now known, with their products localized to different sub-cellular compartments for the regulation of the myriad intracellular membrane trafficking steps, functioning as molecular switches in a myriad of processes. We are interested in the role of a brain-enriched Rab, Rab31 (also known as Rab22B) in astroglia differentiation and function. First shown to regulate membrane traffic between the trans-Golgi network and the endosomes, Rab31 has since been implicated in pathological processes like cancer. We have earlier found that Rab31 is highly expressed in the astroglia lineage throughout development. The role of Rabs in astroglia physiology and pathology is unknown. One hint we had from our earlier analysis is that Rab31 modulates the trafficking of EGF receptor (EGFR) in cancer cells like A431 and HeLa. In this project, we wish to investigate how exactly does Rab31 influence cell fate of neural progenitor cells (NPCs), and secondly determine the roles of Rab31 in astrocyte physiology. We shall check how manipulations of Rab31 levels affect fate of NPCs at the subventricular zone and the subgranular zone of the dentate gyrus. We had also hypothesized that Rab31 could enhance the channelling of internalized EGFR towards the late endosome/lysosome as oppose to a cell surface recycling pathway, which his would alter its signalling profile, and we shall test this hypothesis. We shall also investigate whether Rab31 has a role in modulating the trafficking of other astrocytic surface receptors. EGF has been shown to transactivate nerve growth factor receptor tyrosine kinase (Trk) A and B. We shall therefore ask if Rab31 plays a role in modulating trafficking of TrkA/B and the low affinity neurotrophin receptor p75NTR in astrocytes. We shall also examine whether Rab31 plays a role in the process of astrogliosis during CNS injury. References and readings:

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3. Chua CE and Tang BL (2014) Engagement of the Small GTPase Rab31 and its effector, Early Endosome Antigen 1, is Important for trafficking of ligand-bound Epidermal Growth Factor Receptor from early to late endosome. J. Biol. Chem. 289: 12375-12389. 4. Chua CE and Tang BL (2014) Rab31 is expressed in Neural Progenitor Cells and plays a role in their differentiation. FEBS Lett. 588: 3186-3194. Chua CE and Tang BL (2015) The role of the small GTPase Rab31 in cancer. J. Cell Mol. Med. 19: 1-10. Department of Obstetrics & Gynaecology Principal Investigator

Project Title with a brief description

A/Prof Chong Yap Seng [email protected]

For PhD project Maternal diet during pregnancy and infant neurodevelopment

Co- supervisor: Dr Anne Rifkin [email protected] Dr Mary Chong [email protected]

Omega 3 fatty acids such as docosahexaenoic acid (DHA) and arachidonic acid (AA) are important to foetal and infant growth and development. Deficiencies and imbalances of long chain polyunsaturated fatty acids (LCPUFAS including omega 3 and omega 6 fatty acids) are associated with impairments in cognitive and behavioural performance. While some RCTs during pregnancy suggest that prenatal DHA status might have subtle positive effects on neurodevelopmental and behavior outcomes (Jensen CL, 2005; Cheatham CL, 2011, Drover JR, 2011), these beneficial effects have not been consistently replicated in other good quality studies (Dunstan JA, 2008; Judge MP, 2007; Makrides M, 2010). This is said to be in part due to the methodological limitations of the study and also the genetic heterogeneity in fatty acid metabolism. In addition, compared to their Western counterparts, little is also known about these associations in Asian populations. Research on the associations between maternal dietary patterns during pregnancy and the effects on infant neurodevelopment is also sparse. This research will use data from the GUSTO (Growing Up in Singapore Towards healthy Outcomes) mother-offspring cohort study in Singapore and the largest in Asia. It is a longitudinal study designed to understand the developmental origins of health and disease from an Asian perspective. A sub-cohort of 500 pregnant women were recruited during the first trimester of pregnancy and followed up regularly throughout pregnancy and the nutrition and neurodevelopment of the child tracked from birth to 8 years of age. The existence of the three major ethnic groups (Chinese, Malay and Indians) in the local Singapore population provides an ideal backdrop

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for a study to investigate the associations of maternal omega 3 fatty acids on infant neurodevelopment and how this may be modified by differential polymorphisms across ethnic groups. Research aims: With this, research projects would be undertaken to examine the following hypotheses: 1) What are the associations between maternal plasma LCPUFAs and infant neurodevelopmental outcomes in early life (0 -2 yrs)? 2) Do FADS gene variants among individuals and possibly across ethnic groups modify the effects of maternal plasma LCPUFAs and infant neurodevelopment? 3) What are the associations between maternal dietary patterns and other nutrient intakes on infant neurodevelopmental outcomes in early life (0 -2 yrs)? Department of Orthopaedic Surgery Principal Investigator

Project Title with a brief description

Dr Angelo All [email protected]

Project 1- Induced Human Oligodendrocyte Precursor Cell Transplants for Improved Functionality after Spinal Cord Injury We propose to directly convert adult human fibroblasts into "induced human oligodendrocyte precursor” cells and to study the remyelination capability of these cells both in vitro and in vivo. We will generate patient-specific cells in a shorter time frame than iPS cells, which requires for reprogramming, and also these newly generated cells would have significantly reduced risk of immune rejection compared to even properly differentiated OPs from currently available human iPS cell lines. The goal of this project is to study whether these cells remyelinate demyelinated axons in the hostile micro-environment after SCI in rats. We will also test the myelin production of the cells in a micro-chamber in vitro. Project 2- Hypothermia after Spinal Cord Injury: Early Markers of Recovery Spinal cord injury (SCI) is a devastating condition that can lead to paralysis of the limbs below the injury level. The secondary phase of injury progression comprises inflammation, demyelination and apoptosis, the major pathological factors that have motivated the search for an early neuroprotective therapy. In animal models, primary focus has been on the effects of post-SCI hypothermia on motor behavioral and histological outcomes with only a few studies of the electrophysiological function of descending spinal cord pathways during cooling. Thus there is a critical need for establishing the full benefits of hypothermic neuroprotection after SCI. Unlike other studies, our unique focus is on the hypothermia-induced enhancements of the afferent sensory conduction, a vital function of

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the sensory-motor system, as assessed by multi-limb somatosensory evoked potentials. The cooling will be followed by a single recording session to acquire local field potentials (LFPs) and multi-unit activity (MUA) from the dorsal pathways in the vicinity of the injury, and later the somatosensory evoked potentials (SSEPs), the cortical response waveform upon peripheral stimulation. The acute microelectrode recordings will be used to develop an early statistical marker to identify animals with maximal long-term recovery of SSEPs. The standard motor behavioral scoring (BBB) and histopathology will be used to provide complementary measures of recovery. The goal of this project is to use the contusive rat model of SCI, followed by an early, local cooling of the injured region. Project 3- Transplantation of Conjugated Nanoparticles to Limit Spinal Cord Injury We propose to synthesize and administer conjugated nanoparticles and study their capability of limiting spinal cord injury (SCI) in a rat model. This treatment approach will enable us to spare more healthy axons and neuropathways within the spinal cord parenchyma and slow down progress of injury after trauma. This is important as SCI is a time-sensitive pathology with a very short window of treatment. The goal of this proposal is to generate a new generation of nanoparticles, which will be safe and effective in preventing progress of trauma in our in vivo model of spinal cord injury. Project 4- Crosstalk among Neural Pathways after Incomplete Spinal Cord Injury Spinal Cord Injury (SCI) in the pediatric population is fairly rare but bears huge socioeconomic consequences. Patients with SCI before their adolescence have different mechanisms of injury and have a better neurological recovery potential for incomplete and mild to severe injuries when compared to adults. Although the SCI is usually diagnosed with the use of MRI, often pediatric SCI are without radiological anomaly. There is also no evidence for the use of neuroprotective approaches for the treatment of SCI in children. There is the need for a better determination of the mechanism of SCI at the epicenter of injury. An understanding of the mechanisms that help promote the improved neurological recovery observed in pediatric patients would also enhance future therapies for all SCI patients. This project deals with incomplete SCI and the study of post-contusion electrophysiological changes in the vicinity of the injury. The goal of this proposal is to characterize functional reorganization of neuropathways on the neuronal reconnection formation at the epicenter of contusion induced SCI. Project 5- Behavioral Study after Spinal Cord Injury Spinal Cord Injury (SCI) is a serious condition that severely impairs mobility and quality of life with no treatment options. After injury, some axons are anatomically continued but not functional. These neurons are electrically excitable cells and they are the focus of our Updated in Jan 2018

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treatment in the acute phase. One approach to treat SCI is through limiting and preventing the destruction of healthy neurons in the spinal cord parenchyma, thus aiding functional recovery. A set of motor behavioral assessments, such as BBB open field locomotion test, thermal sensation, rotorod, tactile allodynia and balance beam will be proposed to thoroughly investigate the onset of injury as well as long-term progress of injury. We will study phenotypic outcomes and the possible limitations during the progress of injury. These assessments will be statistically analyzed and compared with the histological examination as well as our previous electrophysiological assessments. Department of Pharmacology Principal Investigator

Project Title with a brief description

A/Prof Bian Jinsong [email protected]

Project 1: Activation of Na+/K+ ATPase (NKA) is a new strategy to treat stroke Stroke is a major cause of disability and death worldwide. Na+/K+ ATPase (NKA) activity is decreased in the brain of stroke patients. We developed an antibody against the DR region of NKA (DR-Ab), which not only stimulated NKA activity but also produced neuroprotective effects against neuronal cell death in ischemia. In the transient middle cerebral artery occlusion (tMCAO) mice, both preand post-stroke treatment with DR-Ab significantly decreased the infarction volume. We therefore hypothesize that DR-region of NKA is a new target to treat ischemic brain diseases and activation of NKA with DR-Ab may be useful to treat stroke. This proposal is therefore designed to study the underlying mechanisms for the therapeutic effect of DR-Ab. Specifically, we will: 1. Confirm the therapeutic effect of DR-Ab on stroke. Time- and dosedependent responses of DR-Ab (intravenous injection) will be studied. 2. Investigate the underlying signaling and molecular mechanisms of the neuroprotective effects of DR-Ab. The effect of DR-Ab on cell apoptosis, membrane NKA loss and excitotoxicity will be studied. Project 2: Hydrogen sulfide is a novel approach to treat neurodegenerative diseases Neurodegenerative disorders represent a major cause of disability and death, with an unmet need for therapies that alter disease progression. Hydrogen sulfide (H2S) has recently been reported to be an important neuromodulator in the brain. Abnormal generation and metabolism of H2S may be actively involved in the pathogenesis of central nervous system (CNS) diseases. We recently reported that both endogenous and exogenous application of H2S produces therapeutic effects on Parkinson’s disease and Alzheimer’s disease. However, the molecular mechanisms are still unclear. In this project, we will examine the effects of H2S on phosphorylation and aggregation of important proteins (e.g. Tau, amyloid β aggregation, STAT3, etc). H2S- induced S-sulfhydration of these proteins will be

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studied. The involved cysteine residues in these proteins will be identified. A/Prof Christopher Chen [email protected]

1) Dementia Neuroimaging We propose to use multi-modal magnetic resonance imaging (MRI) and retinal imaging techniques to systematically study brain functional and anatomical abnormalities and rate of annual changes as well as vascular deficits associated with prodromal dementia (Vascular Cognitive Impairment No Dementia, VCIND), mild VaD, AD. We hypothesise that multi-modality MRI methods and novel retinal imaging will provide novel insights in evaluating brain atrophy and lesions, cerebral artery stenosis, as well as functional deficits associated with different types of dementia and thus identifying dementia-subtype specific imaging markers. Recent publications SJ van Veluw, S Hilal, HJ Kuiff, MK Ikram, X Xin, BY Tan, N Venketasubramanian, GJ Biessels, C Chen. Cortical Microinfarcts on 3T MRI : Clinical Correlates in Memory Clinic Patients. Alzheimer’s & Dementia (accepted) X Xu, S Hilal, SL Collinson, EJY Chong, MK Ikram, N Venketasubramanian, CLH Chen The Association of Magnetic Resonance Imaging Markers of Cerebrovascular Disease Burden and Cognition Stroke (accepted) S. Hilal, M.A. Shaik, N. Venketasubramanian, W.J. Niessen, H. Vrooman, T.Y. Wong, C. Chen, M.K. Ikram Subcortical atrophy in cognitive impairment and dementia J Alzheimer’s Disease (accepted) CY Cheung,,YT Ong, S Hilal, MK Ikram, S Low, YL Ong, N Venketasubramanian, P Yap,D Seow, CLH Chen, TY Wong. Retinal ganglion cell analysis using high-definition optical coherence tomography changes in patients with mild cognitive impairment and Alzheimer’s Disease J Alzheimer’s Disease 45 (2015) 45-56 S Hilal, M Saini , CS Tan, JA Catindig, ,YH Dong , RL Hollandez, WJ Niessen, HA Vrooman, E Ting, TY Wong, C Chen, N , Venketasubramanian MK Ikram. Intracranial stenosis, cerebrovascular diseases and cognitive impairment in Chinese Alzheimer Disease & Associated Disorders 29 (2015) CY Cheung,,YT Ong, MK Ikram,, SY Ong, X Li, S Hilal, JA Catindig, N Venketasubramanian, P Yap,D Seow, CP Chen, TY Wong Microvascular network alterations in the retina of patients with Alzheimer’s Disease Alzheimer's and Dementia 10 (2014) 135-42 2) Clinical Trials in Dementia and Stroke Our current focus is on developing a regional network for innovative trials into conditions prevalent and pertinent to Asia. We also plan to enhance Singapore's capability for conducting increasingly sophisticated early phase studies utilizing biomarkers such as neuroimaging.

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Recent publications include N Venketasubramanian, SHY Young, SS Tay, T Umapathi, AY Lao, HH Gan, AC Baroque II, , JC Navarro, HM Chang, JM Advincula, S Muengtaweepongsa, BPL Chan, CL Chua, N Wijekoon, HA de Silva, JHB Hiyadan, NC Suwanwela, KSL Wong, N Poungvarin, GB Eow, CF Lee, CLH Chen. CHInese Medicine NeuroAiD Efficacy on Stroke recovery – Extension Study (CHIMES-E : A multicenter study of longterm efficacy Cerebrovascular Diseases 39 (2015) 309-18 J Navarro, H Gan, A Lao, A Baroque, JH Hiyadan, C Chua, MC San Jose, J Advincula, CF Lee, MG Bousser, C Chen. Baseline Characteristics and Treatment Response of Patients from the Philippines in the CHIMES study. International Journal of Stroke SA100 (2014) 102-5 CLH Chen, N Venketasubramanian, CF Lee, KSL Wong, MG Bousser for the CHIMES Investigators. Effects of MLC601 on Early Vascular Events in Post-stroke Patients – The CHIMES Study Stroke 44 (2013) 3580-3583 CLH Chen, SHY Young, HH Gan, R Singh, AY Lao, AC Baroque II, HM Chang, JHB Hiyadan, CL Chua, JM Advincula, S Muengtaweepongsa, BPL Chan, HA de Silva, S Towanabut, NC Suwanwela, N Poungvarin, S Chankrachang, KSL Wong, GB Eow, JC Navarro, N Venketasubramanian, CF Lee, M-G Bousser, for the CHIMES Study Investigators Chinese Medicine Neuroaid Efficacy on Stroke Recovery A Double-Blind, Placebo-Controlled, Randomized Study Stroke 44 (2013) 2093-2100 3) Biomarker Discovery in Stroke and Dementia Investigating the pathological basis and biomarkers for stroke and vascular cognitive impairment utilising tissue samples from well characterized clinical cohorts on platforms including proteomics, lipodomics and a candidate biomarker approach Recent publications K Narasimhalu, J Lee, YL Leong, L Ma, DA De Silva, MC Wong, HM Chang, C Chen Inflammatory markers and their association with post stroke cognitive decline International Journal of Stroke 10 (2015) 513-8 S Hilal, YL Chai, MK Ikram, S Elangovan, BY Tan, X Xin, JY Chong, N Venketasubramanian, AM Richards, JPC Chong, MKP Lai, C Chen. Markers of Cardiac Dysfunction in Cognitive Impairment and Dementia. Medicine 94 (2015) e297 Adav S, Qian J, Ang Y L, Kalaria R, Lai M, Chen C, Sze SK. iTRAQ Quantitative Clinical Proteomics Revealed Role of NaK-ATPase and its Correlation with Deamidation in Vascular Dementia Journal of Proteome Research 13 (2014) 4635-46 Datta A, Chen CP, Sze SK Discovery of Prognostic Biomarker Candidates of Lacunar Infarction by Quantitative Proteomics of Exosome Enriched Plasma PLOS-1 9 (2014) e94663 Lam SM, Wang Y, Duan X, Wenk MR, Kalaria RN, Chen CP, Lai MK, Shui GH. The brain lipidomes of subcortical ischemic vascular dementia and mixed dementia Neurobiology of Aging 35 (2014) 2369-2381 Datta A, Qian J, Chong R, Kalaria RN, Francis PT, Lai MK, Chen Updated in Jan 2018

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CP, Sze SK. Novel Pathophysiological Markers are Revealed by iTRAQ-based Quantitative Clinical Proteomics Approach in Vascular Dementia. Journal of Proteomics 99C (2014) 54-67 4) Animal Models and Mechanisms of Dementia Investigating novel shared mechanisms between Alzheimer’s Dementia and Diabetes by use of cell cultures and animal models of hyperglycemia and AD. Investigating novel treatments for dementia. Recent publications YA Lim, LA Murray, MKP Lai, C Chen NeuroAiD™ (MLC601) and Amyloid Precursor Protein Processing Cerebrovascular Diseases 35 (2013) 30-37 Götz J, Lim YA, Eckert A. Lessons from two prevalent amyloidoseswhat amylin and Aβ have in common. Front Aging Neurosci. 2013 Lim YA, Rhein V, Baysang G, Meier F, Poljak A, Raftery MJ, Guilhaus M, Ittner LM, Eckert A, Götz J. Abeta and human amylin share a common toxicity pathway via mitochondrial dysfunction. Proteomics. 2010 10(8):1621-33 5) Neurochemistry and Molecular Pathology of Dementia Investigating the neurochemistry molecular pathology and their clinical correlates in dementia using tissue samples from well characterized clinical cohorts. Recent publications Howlett DR, Whitfield D, Johnson M, Attems J, O’Brien J, Aarsland D, Lai MK, Lee JH, Chen CP, Ballard C, Hortobágyi T, Francis PT. Regional Multiple Pathology Scores are Associated with Cognitive Decline in Lewy Body Dementias. Brain Pathology 25 (2015) 401-8 Mohamed NE, Lee JH, Francis PT, Esiri MM, Chen CP, Lai MK. Differential alterations of neocortical GluNreceptor subunits in patients with mixed subcortical ischemic vascular dementia and Alzheimer’s disease. J Alzheimer’s Disease 44 (2015) 431-37 X Gallart-Palau, A Serra, J Qian, CP.Chen, RN Kalaria, SK Sze Temporal lobe proteins implicated in synaptic failure exhibit differential expression and deamidation in vascular dementia. Neurochemistry International 80 (2015) 87-98 NE Mohamed, JH Lee, PT Francis, D Aarsland, CG Ballard, CP Chen, MKP Lai. Loss of NMDA NR1 subunit immunoreactivity correlates with dementia severity in Lewy Body dementias J Neural Transm 121 (2014) 71-8 MG Tan, CL Lee, JH Lee, PT Francis , R Williams, C Chen, PTH Wong, MJ Ramírez, MKP Lai. Decreased rabphilin 3A immunoreactivity in Alzheimer’ Disease is associated with Aβ burden Neurochemistry International 64 (2014) 29-36 6) The Neuropsychology Impairment

of

Dementia

and

Cognitive

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longitudinal clinical studies, the multi-ethnic Epidemiology of Dementia in Singapore study and studies in primary health care. Recent publications QL Chan, X Xu, MA Shaik, SST Chong, RJY Hui, CLH Chen YH Dong Clinical utility of the informant AD8 as a dementia case finding instrument in primary healthcare J Alzheimer’s Disease (accepted) X Xu, SL Ang, S Hilal, QL Chan, TY Wong, N Venketasubramanian, MK Ikram, CLH Chen Association of neuropsychiatric symptoms and sub-syndromes with cognitive impairment in community-dwelling Asian elderly. Int Psychogeriatrics (accepted) JHQ Shen, Q Shen, H Yu ,JS Lai, JL Beaumont, Z Zhang , H Wang , S Kim, C Chen, T Kwok, SJ Wang, D Lee, J Harrison, J Cummings. Validation of an Alzheimer’s Disease Assessment Battery in Asian Participants With Mild to Moderate Alzheimer’s Disease Am J Neurodegener Dis 3 (2014) 158-69 Dong Y, Slavin MJ, Chan BP, Venketasubramanian N, Sharma VK, Collinson SL, Sachdev P, Chen CL. Improving screening for Vascular Cognitive Impairment at 3-6 months after Mild Ischemic Stroke and Transient Ischemic Attack. International Psychogeriatrics 26 (2014) 787-93 A/Prof Gautam Sethi [email protected]

NF-κB and STAT3 are ubiquitously expressed transcription factors involved in cell survival, adhesion, inflammation, and differentiation. NF-κB and STAT3 are constitutively active in most cancers and responsible for the transcription of various genes involved in initiation, promotion, and perpetuation of cancer. Further, these proteins can be activated by most cancer treatment modalities and mediates treatment resistance. Hence, the drugs that can suppress NF-κB/STAT3 activation have a great potential as effective therapeutics for treatment of various solid cancers and hematological malignancies. Our group is therefore interested in investigating the following major aims: 1. Molecular mechanism(s) of activation of transcription factors NFκB/STAT3 by carcinogens, tumor promoters, infectious and inflammatory agents. 2. How tumor cells express constitutive active NF-κB/STAT3, evade apoptosis and develop resistance to chemotherapeutic agents and radiation? 3. Identification of novel pharmacological compounds that can suppress dysregulated NF-κB/STAT3 activation in tumor cells/animal models and patient samples. 4. Potential use of these novel compounds to overcome chemoresistance and radioresistance in xenograft and transgenic cancer models.

A/Prof Gavin Dawe [email protected]

Electroceuticals for stress and metabolic disorders Electroceuticals, the therapeutic application of electronic devices interfacing with nerves/organs, is a new frontier between medicine and engineering. Electroceuticals enable many advantages over

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pharmaceuticals including: (1) targeting of specific nerves/organs, reducing adverse effects compared to systemic drugs acting throughout the body; (2) continuous treatment adjustable in response to patient needs or by closed-loop feedback of physiological parameters, improving therapeutic compliance and clinical outcome over the fixed intermittent dosing typical of pharmaceuticals; and (3) highly reversible treatment allowing immediate cessation by switching off the device. Working together with the Singapore Institute of Neurotechnology (SINAPSE), who are world leaders in developing a portfolio of microelectronic technologies enabling peripheral nerve stimulation and recordings experiments that were never before possible, this project will explore novel strategies for modulation of autonomic nervous system function to regulate stress and metabolic disorders. Novel signaling functions of amyloid precursor protein Amyloid precursor protein (APP) is best known for its sequential cleavage by secretase enzymes to release the amyloid beta peptide that aggregates as amyloid plaques in Alzheimer’s disease. However, APP is a complex transmembrane protein with many different functional domains. We have characterized a number of novel functions of APP. This project will discover new signaling functions of APP and explore their potential implications in health and disease. Example publications (*corresponding author): Liu C, Tan FC, Xiao ZC, Dawe GS*. Amyloid precursor protein enhances Nav1.6 sodium channel cell surface expression. J Biol Chem. 2015 May 8;290(19):12048-57. Shu R, Wong W, Ma QH, Yang ZZ, Zhu H, Liu FJ, Wang P, Ma J, Yan S, Polo JM, Bernard CC, Stanton LW, Dawe GS*, Xiao ZC*. APP intracellular domain acts as a transcriptional regulator of miR-663 suppressing neuronal differentiation. Cell Death Dis. 2015 Feb 19;6:e1651. Xu DE, Zhang WM, Yang ZZ, Zhu HM, Yan K, Li S, Bagnard D, Dawe GS*, Ma QH*, Xiao ZC*. Amyloid precursor protein at node of Ranvier modulates nodal formation. Cell Adh Migr. 2014;8(4):396-403. Ma QH, Bagnard D, Xiao ZC*, Dawe GS*. A TAG on to the neurogenic functions of APP. Cell Adh Migr. 2008 JanMar;2(1):2-8. Ma QH, T Futagawa T, Yang WL, Jiang XD, Zeng L, Takeda Y, Xu RX, Bagnard D, Schachner M, Furley AJ, Karagogeos D, Watanabe K, Dawe GS*, Xiao ZC*. A TAG1-APP signalling pathway through Fe65 negatively modulates neurogenesis. Nature Cell Biology 2008; 10(3):283-94. The role of the nucleus incertus and relaxin-3/RXFPs in the neurobiology of appetite Relaxin-3, the most recently discovered member of the insulin/insulin-like growth factor/relaxin peptide family, is expressed Updated in Jan 2018

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as a neuropeptide in the brain where its cognate receptor is relaxin/insulin-like family peptide receptor 3 (RXFP3). The principal source of relaxin-3 is the brainstem nucleus incertus. The nucleus incertus projects widely to midbrain and forebrain structures, including hypothalamus, amygdala, medial septum, hippocampus and prefrontal cortex. The nucleus incertus highly expresses corticotropin releasing hormone receptor type 1 and these ascending projections are activated in stress responses. This has led to the proposition that the nucleus incertus/relaxin-3 system, like the raphe serotoninergic and locus coeruleus noradrenergic ascending projection systems, is a novel ascending projection system implicated in modulation of emotional and cognitive function by arousal and stress responses. Emerging roles for nucleus incertus relaxin-3/RXFP3 networks in feeding, arousal, stress and memory suggest novel targets for the treatment of depression and other neuropsychiatric diseases. This project will explore the neuronal circuitry and functions of the nucleus incertus and relaxin-3/RXFP3 in the neurobiology of appetite. Dr Mitchell Lai [email protected]

Bioactive compounds for anti-neuroinflammation therapy Unregulated neuroinflammation is pathogenic for certain diseases (e.g., bacterial encephalitis) and exacerbates other diseases (e.g., stroke and Alzheimer’s disease). In this project, you will work with a number of naturally derived compounds which show potential as antineuroinflammatory therapeutics. You will use cellular and animal neuroinflammation models and a combination of pharmacological, molecular biology and cell biology techniques to study their mechanism of action, cellular targets and potential clinical efficacy. Neurochemistry of post-stroke dementia Dementia occurs in a proportion of patients who suffered initially mild strokes and is a cause of significant morbidity, but the neurochemical changes which underlie the loss of cognitive abilities after stroke have not been elucidated. In this project, you will work with animal models as well as postmortem brain tissues to uncover changes in various neurotransmitter systems after a stroke / hypoxic event. It is anticipated that this knowledge will eventually lead to improved management and outcomes for post-stroke dementia.

A/Prof Low Chian Ming [email protected]

Project 1: Characterization and pharmacological intervention in chemoagent-induced peripheral neuropathic pain. (Co-supervisors: A/P S. Khanna, Dept of Physiology; Prof Tat-Leang Lee, Dept of Anaesthesia) Chemotherapy drugs such as platinum analogues (oxaliplatin, cisplatin), vinca alkaloids (vincristine, vinblastine), taxanes (paclitaxel, docetaxel) and thalidomide are often used in the treatment of cancer. However, the neurotoxicity of these drugs result in patients developing side effects such as peripheral neuropathy with pain severity ranging from mild and dysesthetic to excruciating. This

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painful side effect could be chronic thus affecting patients’ quality of life. This project aims 1) to characterize the pathway of paclitaxel-induced peripheral neuropathic pain rat model developed in our laboratories and 2) to investigate pharmacological tools to alleviate the neuropathic pain. With the rising cancer rates in Singapore, better understanding of the pathogenic mechanism can bring about better treatment options alleviating chemotherapy-induced peripheral neuropathy. Project 2: Neuroinflammation in neurodegenerative diseases (Co-supervisors: A/P Yulin Lam, Dept of Chemistry; Prof. Chandra Verma, Bioinformatics Institute, A*STAR) Our collaborator and my laboratory have shown neuroinflammation precedes classical Alzheimer’s disease (AD) pathologies. We had identified the mediator triggering neuroinflammation. We further demonstrated the beneficial effects of preventing neuroinflammation led to delay onset of AD in cdk5/p25 transgenic mouse model. In collaboration with medicinal chemists from Department of Chemistry, NUS, and molecular modellers from BioInformatic Institute, A*STAR we embarked on drug design and discovery on known scaffold that has shown to inhibit the target responsible for the release of pro-neuroinflammatory mediator. We have successfully generated more potent analogues. This project aims to test our new analogues on cell-based assay and neurodegenerative animal model. Dr Michelle GK Tan [email protected] [email protected]

Isoform of Fyn tyrosine kinase as a novel potential therapeutic target for Alzheimer’s disease Fyn tyrosine kinase has been suggested to have a pathogenic role in Alzheimer's disease (AD). Targeting Fyn as a therapeutic approach has been proposed, but may be limited by adverse effects given Fyn’s regulatory role in neuronal functions. We observed that alternatively spliced FynT isoform is associated with two neuropathological features of AD: neurofibrillary tangles and reactive astrogliosis, suggesting that isoform-specific up-regulation of FynT may be involved in neuronal degeneration and astrocyte-mediated neuroinflammation in AD. In this project, we propose a multimodal study using cell-based assays, animal model and postmortem brain tissue to test our hypothesis. In addition to providing further insight into the pathogenic role of Fyn in AD and other neurodegenerative dementias, successful identification of FynT as a novel target will drive therapeutic approaches which specifically target FynT to potentially improve drug efficacy and reduce off-target effects. To study the role of microRNA dysregulation in modulating abnormal increase of inositol triphosphate 3-kinase B (ITPKB) expression in Alzheimer's disease

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We demonstrated that inositol triphosphate 3-kinase B (ITPKB) expression was significantly increased in Alzheimer's disease (AD) brain, consistent with previous reports. One of the known functions of ITPKB is to convert inositol 1,4,5-triphosphate (IP3) into inositol 1,3,4,5-tetrakisphosphate (IP4), regulating calcium signaling downstream of phospholipase C. Recent study showed that overexpressing ITPKB transgene in AD mouse model exacerbates disease pathology through activation of ERK1 signaling pathway. However, the mechanism of abnormally increased ITPKB expression in AD brain is still unknown. Using integrated microRNA and gene expression profiling data, we have identified ITPKB being a potential gene target of six AD down-regulated microRNAs. In this project, we aim to study the role of microRNA dysregulation in modulating ITPKB expression in AD. Department of Physiology Principal Investigator

Project Title with a brief description

Dr Adam Claridge-Chang [email protected]

The role of serotonin in defense behavior in vinegar fly Humans experience emotions as subjective feelings. The emotional states underlying such feelings have a powerful influence on behavior and perception. Human emotional behaviors have closely similar expressions in animals and emotion-related modulatory brain molecules influence animal and human behavior alike. Such evolutionarily conserved similarities reveal a relationship between human and animal brain function that permits the use of animals as research models of human emotional states. An important emotionrelated modulator is 5-hydroxytryptamine (5-HT), also known as serotonin. Serotonin has been implicated in a range of emotional states, however, there remains conflicting and largely incomplete information about its role. Some data connect it to negative emotions including anxiety promotion, while other results connect it to reward, positive emotions and anxiety reduction. We have developed behavioral assays using the genetically tractable black-bellied vinegar fly (Drosophila melanogaster). One assay examines the propensity of flies to follow walls in a chamber; it shows striking genetic similarities to anxiety-related defense behavior assays in rodents. A cluster of serotonin neurons strongly decreases defensive wall following through specific serotonin receptors and propose experiments to further elucidate the neuronal and molecular mechanisms of this effect. We aim to characterize the receptorbearing cells that mediate the wall following changes. A mechanistic understanding of the 5-HT cells that regulate defense behavior in the fly will guide experiments in the costlier, slower vertebrate systems. Findings from the proposed work would similarly guide the work of scientists studying mood and emotional disorders in humans. Positive emotional states in fly and serotonin signalling Humans experience emotions as subjective feelings. The emotional

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states underlying such feelings have a powerful influence on behavior and perception. Human emotional behaviors have closely similar expressions in animals and emotion-related modulatory brain molecules influence animal and human behavior alike. Such evolutionarily conserved similarities reveal a relationship between human and animal brain function that permits the use of animals as research models of human emotional states. An important emotionrelated modulator is 5-hydroxytryptamine (5-HT), also known as serotonin. Some results connect serotonin to reward, positive emotions and anxiety reduction. We have developed an assay in Drosophila melanogaster that measures the extent to which a fly is attracted to the synthetic activation of cells; we find that serotonin neuron activation is strongly attractive. We propose experiments to elucidate the cells involved in the attraction and their target cells. A mechanistic understanding of the 5-HT cells that regulate positive valence in the fly will guide experiments in the costlier, slower vertebrate systems. Findings from the proposed work would similarly guide the work of scientists studying mood and emotional disorders in humans.

A/Prof Christopher Beng-Ti ANG Sialylation and Tumor Cell Invasion in Glioblastoma [email protected] Brain tumors are among the most devastating of cancers, with a mean survival period of just 15 months upon. Currently, clinicians rely on mainly histology to guide treatment decisions. In line with international efforts, we recently showed that resistance to current chemotherapeutic drugs and tumor cell invasiveness can be attributed to a self-renewing, cellular fraction within the tumor mass. This tumor heterogeneity is the reason that brain tumors are notoriously difficult to treat. We further showed that patient genetic profiles can be mapped to such gene activation profiles enriched for stem-like properties. Our proposal here will focus on one such important gene activation program, regulated by a sialylation enzyme, ST3Gal1. We recently demonstrated that ST3Gal1 expression marks a persistent cellular fraction, and is highly enriched in mesenchymal molecular features, typical of aggressive and recurrent tumors. Our orthotopic animal model established from patient cells further demonstrated that ST3Gal1 knockdown abrogated tumor-initiating and -sustaining capability. Moving forward, we will focus on signaling mechanisms that mediate ST3Gal1 activity. Our lab has expertise to interrogate patient clinical databases, and we identified several methylation hallmarks that potentially regulate ST3Gal1. This is a significant List of research projects available for prospective graduate students Updated in Aug 2015 Page 49 endeavor as DNA methylation accounts for up to 70% of tumor profiles. We will test such methylation hallmarks in vitro and in animal models. Our lab deals with several techniques such as primary cell culture, biochemical and molecular biology approaches and flow cytometry. Histology forms a significant thrust of our analyses, along with mouse models that recapitulate the patient Updated in Jan 2018

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heterogeneous spectrum. Our study is unique as we rely on patient genomic content to guide our bench experiments. The successful candidate will be exposed to cross-disciplinary approaches to address clinically important questions in glioma biology. Prof Carlos Ibanez [email protected]

Growth Factor Receptor Signaling and Biology in Nervous System Development and Metabolic Regulation With a focus on growth factor receptor signalling, the aim of this research is to discover novel, general biological principles and mechanisms important for nervous system development and metabolic regulation. We plan to concentrate the main thrust of our future work on two programmes that complement and expand current areas of expertise. The first one focuses on the mechanisms of action and biological activities of the death receptor p75NTR (p75 neurotrophin receptor), and reflects our continued interest in neuronal growth factors, their signalling mechanisms and their potential for the development of novel therapies for neurodegenerative diseases and neurotrauma. This is a program that involves signal transduction studies, mouse genetics approaches, and a drug discovery effort to identify novel p75NTR inhibitors. The second program focuses on the role of a novel regulatory network, formed by members of the transforming growth factor-beta (TGF-) superfamily and their receptors, in the control of glucose homeostasis, fat accumulation and energy balance. It is based on our original discovery of the TGF- superfamily receptor ALK7 and our recent studies on the functions of activin signalling through ALK7 and ALK4 in pancreatic islets, adipose tissue, and hypothalamus. This program offers a new inroad to the study of metabolic regulation and involves complementary studies in cell culture as well as conditional and knock-in mouse models.

Dr John Chua Jia En [email protected]

Mechanisms of protein trafficking to the presynapse An individual neuronal synapse comprises of hundreds of proteins. The formation of synaptic machineries required for synapse establishment and function critically depends on the delivery of these proteins by the intracellular trafficking machinery. Defective trafficking contributes to the development and progression of neurodegenerative disorders such as Alzheimer’s and Huntington’s diseases. We are interested in understanding specific mechanisms and regulatory pathways governing this process. In this project, we aim to learn more about how different groups of presynaptic proteins are directed to synapses and how this process contributes to synaptic development and function. In doing so, we aim to advance our understanding towards how trafficking defects contribute to the etiology of neurodegenerative disorders. Molecular techniques, primary cell culture and live imaging of neurons are some of the techniques to be applied.

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Dr Karthik babu Mallilankaraman Genome-wide RNAi screening to discover novel modulators of endothelial cell death and their molecular mechanism of action. [email protected] Reperfusion of the ischemic tissue is a major trigger and contributor of cell death in a variety of cardiovascular, metabolic and neurological diseases. Cells and tissues exposed to hypoxia during ischemia may sustain damage and cell death upon reoxygenation by restitution of blood flow. This Hypoxia/reoxygeneation (H/R)-related injury has long been associated with the enhanced production of reactive oxygen intermediates generated during reoxygenation. Additionally, H/R may cause injury through the recruitment of proinflammatory leukocytes. Endothelial cells represent the first cell type injured by reactive oxygen species generated during H/R as these cells form the first line of defense in the vasculature. In vitro, the mechanisms of endothelial cell death caused by H/R remain poorly understood and may involve both necrotic and apoptotic forms of cell death. Recent advances in the genomics have rendered researchers to develop various tools to dissect molecular pathways in complex diseases. Specifically, the discovery of RNA interference (RNAi) and the rapid development of technologies exploiting its functional applications have enabled us to examine the consequences of lossof-function of a particular gene product in mammalian cells. Completion of human genome project further advanced the field towards developing genome-wide RNAi libraries targeting the entire human genome. Genome-wide RNAi libraries have made it possible to carry out large scale, phenotype-based screens, which have yielded potent information on diverse cellular processes ranging from oncogenic to infectious diseases. Nevertheless, such screening methodologies have never been employed in vascular diseases. We have developed an unbiased genome-wide screening approach to identify novel modulators of cell death. My lab will use this approach to identify previously unknown molecular pathways that exist in endothelial cells during Ischemia/Reperfusion injury. Dr LING Shuo-Chien [email protected]

Mechanisms of Neuronal and Synaptic Dysfunctions in Ageing and Neurodegeneration Aging poses both a fascinatingly biological question and a growing medical problem as aging is the leading risk factor for ageassociated diseases, many of which are late adult-onset neurodegenerative diseases, including Alzheimer’s disease (AD), Parkinson’s disease (PD), Frontotemporal dementia (FTD), and Amyotrophic Lateral Sclerosis (ALS, also known as motor neuron disease). The current goal of my lab is to define and understand the processes underlying normal and pathological brain aging by elucidating the underlying molecular and cellular mechanisms and how perturbation of the at-risk systems cause premature failing that leads to neurodegeneration. The overall strategy is to use disease-related genes as

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molecular handles to probe the gene-phenotype relationship systematically in mice that are genetic mimics of human diseases during aging and neurodegeneration. Specifically, we focus on two RNA-binding proteins, TDP-43 (TAR DNA-binding protein 43 KDa) and FUS/TLS (fused in sarcoma/translocated in liposarcoma), whose mutations are causal for ALS and FTD and inclusions of both proteins are the defining pathological hallmarks in the majority of ALS and FTD patients. We will interrogate molecular and synaptic changes in selective brain regions and in neuronal populations as well as the contribution of non-neuronal neighboring cells by combining mouse genetics with genome-wide quantitative methodologies. In addition, we also investigate how dysfunction in autophagy (i.e., self-eating) contributes to neurodegeneration. We also collaborate with Bioengineering groups for high-throughput nanotechnology and microfluidic system to advance the technology for biomarker development and detection. By integrating neuroscience with leading edge technology, we expect that we will identify quantitative changes in gene expression that are critical for initiating and/or accelerating age-dependent neurodegeneration. The long-term goal of our laboratory is to revert the molecular and cellular alternations to rescue neuronal damages and ultimately to slow down aging and subsequent neurodegeneration. Prof Edward H. KOO [email protected]

Molecular mechanisms of neuronal injury or neurodegeneration The Laboratory for Molecular Neurodegeneration at YLL School of Medicine was established by Dr. Ling Shuo-Chien and Prof. Edward Koo in 2013. Our research focus is on the mechanisms of neurodegenerative diseases, a group of brain disorders that is often age-associated and includes Alzheimer and Parkinson diseases, amyotrophic lateral sclerosis, frontotemporal dementia, among others. We take a cell and molecular biology approach to our studies with an emphasis on animal models and translational research. My laboratory’s philosophy is to study both the “normal and abnormal biology” of genes and proteins that are implicated in disease pathogenesis. In many instances, mutations are found in genes with unknown function and which require thorough investigations of the basic functions of these genes and proteins before we can appreciate how their dysfunction contributes Alzheimer disease where we have concentrated on both basic and translational studies as well as experimental therapeutics. Ongoing and future projects include: • • • • • •

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Mechanisms of synaptic injury in Alzheimer disease Role of caspase cleavage of the amyloid precursor protein (APP) in neurodegeneration Mechanisms of tau mediated synaptic dysfunction Investigations into mechanisms of neuronal vulnerability in Alzheimer disease Contribution of vascular dysfunction to neurodegeneration Neurobiological basis of cognitive loss due to chemotherapy Page 66

treatment in cancer patients (“chemobrain”) A/Prof Sanjay Khanna [email protected]

Memory networks and chronic pain: A comparison of the neural, cellular and molecular basis of fear and pain-induced aversive memories Recently, we made a salient observation that a common pool of neurons in the forebrain medial septum mediate both foot shockinduced fear and pain-induced aversion. The effect on aversion was related to the regulation of cellular mechanisms in the anterior cingulate cortex that confer an aversive value to pain. However, at present it not clear whether fear and pain represent distinct or continuum of encoding of unpleasant experiences by a common memory network. Using behavioural, electrophysiological, optogenetic and cellular/molecular techniques, the present proposal will test the hypothesis that overlapping mechanisms in medial septum and selected forebrain regions modulate fear and paininduced aversion. Here it is notable that pain, especially chronic pain, is debilitating and alters the life style of patients and is an area of enormous unmet clinical need because rational treatment options are limited. The successful outcome of the present study will advance the knowledge about the physiological and neurochemical basis of pain and suggest novel neural and neurochemical targets for the control of the aversive state of the chronic pain.

Department of Psychological Medicine A/Prof Rathi Mahendran [email protected]

A Randomised Control Trial of Art Therapy and Music Reminiscence in Community-Living Elderly with Mild Cognitive Impairment Tentative start date; March to May 2016. Patient recruitment Jan 2016. Patients with cognitive impairment commonly experience neuropsychiatric symptoms that diminish their quality of life. Earlier naturalistic studies by the Department of Psychological Medicine team established that psycho-social interventions (Mindfulness Practice, Music Reminiscence, Art Therapy) can bring about significant changes at the functional and molecular levels in the elderly. A Randomized Control Trial on the efficacy of Mindfulness Practice versus Health Education Programs was completed in July 2015. This study scheduled for early 2016, will now compare the efficacy of Art Therapy and Music Reminiscence Activity with a Control Group. Elderly persons with MCI confirmed by Neuropsychological Tests will be randomized into 3 arms (N=105; 35 in each arm). The two active arms will receive Art Therapy (including monthly Gallery Visits) and Music Reminiscence Activity respectively, weekly for 12 weeks then fortnightly for 6 months. The Control arm will not be involved in any activities but will be provided self-help reading materials. Assessments will include Neuroimaging (fMRI) and EEG,

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Neuropsychological tests and blood investigations for chromosome and telomere studies, inflammatory and oxidative stress markers. Secondary measures will include Life Satisfaction, Quality of Life, assessments of mood using GDS and GAI. Assessments will be done at Baseline, 3 months and 9 months.

Category: Functional Ageing Department of Biochemistry Principal Investigator

Project Title with a brief description

A/Prof Yew Wen Shan [email protected]

Current Synthetic Enzymology research interest focuses on protein engineering and biocatalysis, with emphasis on using structural and mechanistic enzymological knowledge to develop therapeutics. We are involved in the following research areas: 1. Defining Synthetic Enzymology as an enabling platform for the

2.

3. 4. 5. 6.

Construction of Next-Generation Synthetic Biology solutions for Pharma and Consumer Businesses. Using Synthetic Enzymology to discover and develop novel therapeutic biomolecules, in the areas of infectious diseases, metabolic disorders, cancer and aging. Developing anti-virulence Quorum-Based Technology for use in the biomedical industry. Using Synthetic Lixiviant Enzymology for Biomining of Electronic Wastes for Environmental Sustainability. Development of Lead Compounds Targeting Enzymes involved in Metabolic Cancer. Using Synthetic Enzymology for the Construction of Therapeutic (Probiotic) Cells for the Treatment of Metabolic Diseases and Infectious Diseases.

Department of Psychological Medicine Principal Investigator

Project Title with a brief description

Dr Feng Lei [email protected]

The association between diet and health status in Asian elderly: a cross-sectional study The Diet and Healthy Ageing (DaHA) study aims to examine the associations between diet and physical, affective and cognitive functioning in Asian elderly. The dietary factors to be examined include: (1) dietary patterns; (2) macro and micronutrients intake; (3) selected food and beverage items; (4) nutritional biomarkers One thousand two hundred elderly persons aged 60 and above will be recruited from a geographically defined area in Jurong, Singapore. Information on diet, self-rated health, cognitive function, depressive symptoms, anxiety symptoms, sleep quality and physical

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performance will be collected. Fecal samples will be collected for the analysis of gut microbiota. Venous blood samples will be collected for diagnostic laboratory tests and the measurement of telomere length and nutritional biomarkers. Multiple linear regressions and multiple logistic regressions will be performed to examine the associations between various dietary variables and health variables. Choral singing for the prevention of dementia: a randomized controlled trial Aims: To evaluate the efficacy and mechanisms of choral singing in the prevention of dementia and examine the underlying neural mechanisms of the expected efficacy using Magnetic Resonance Imaging (MRI) technique. Hypotheses: Choral singing could prevent cognitive decline among community-dwelling elderly who are at high risk of dementia. The underlying neural mechanisms involve changes in brain structure and function that can be quantified using MRI technique. Methods: Study design: randomized controlled trial. Inclusion criteria: (1) Community-living elderly aged 60 years and above; (2) Subjective cognitive complaints based on self-report; (3) Objective cognitive impairment based on neuropsychological test scores (Z score -1.5); (4) Multiple risk factors of dementia such as family history, depression, etc.; (5) Not demented: Clinical Dementia Rating (CDR) global score=0. Treatment Arms: (1) Intervention arm: weekly choral singing for 2 years; (2) Control arm: weekly general health education and group activities for 2 years. Intervention period and follow-ups: 24 months (a ten year follow-up will be planned in the future). Primary outcome measures: (1) Cognitive decline measured by the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment (MoCA); (2) Test scores from a standard neuropsychological test battery: Rey Auditory Verbal Learning Test (RAVLT), Digit Span, Color Trails Test, Block Design, Verbal Fluency, Symbol Digit Modality Test (SDMT). Secondary outcome measures: (1) Brain magnetic resonance imaging (MRI); (2) Depressive symptoms; (3) Anxiety symptoms; (4) Neuropsychiatric symptoms; (5)Perceived stress. Fasting venous blood samples will be collected, processed and stored for future analysis. Significance: The study will provide important data on the efficacy and underlying mechanisms of choral singing in the prevention of dementia. Evidence-based choral singing intervention for dementia prevention will have enormous potential impact for healthcare savings and societal welfare. Aging in a Community Environment Study (ACES) In the Aging in a Community Environment Study (ACES), we will recruit a cohort of community dwelling elderly to study aging and health longitudinally. The aims of the study are: 1. To study the markers and predictors of healthy and functional aging in a community environment. 2. To describe the trajectories of cognitive and physical function decline in normal and abnormal aging. Updated in Jan 2018

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3. To identify modifiable risk factors for the prevention of dementia, frailty and disability. The ACES will recruit subjects from the Diet and Healthy Aging (DaHA) study and expand study population through door-to-door recruitment in Jurong, Singapore. The DaHA study has recruited over 900 subjects from a geographically defined area that surrounds the Jurong Point shopping mall. The ACES will expand existing subject pool so the cohort will have 50% Chinese, 25% Malay, 25% Indian, with a total of 1200 community-living seniors in the natural observational cohort for regular follow-up assessments and further studies. Trained research staffs will conduct face-to-face interview and assessment to collect a wide range of data related to health and functioning in aging. Blood, urine and stool will be collected for the analysis of biological markers. Imaging, diagnostic assessment and further biosampling will be conducted on selected subgroup of subjects from the cohort. Dr Alfredo Franco-Obregón [email protected]

Recapitulating the metabolic and physical adaptations of exercise with brief pulsed electromagnetic field therapy Exercise benefits our overall health and well-being. Skeletal muscle, as our largest tissue mass, has evolved to play a major homeostatic role in whole body maintenance. Physical activity originates from, as well as impinges on, skeletal muscles, causing them to release a myriad of autocrine, paracrine and endocrine trophic and regenerative agents (myokines) with broad systemic benefits. Consequently, there has been much recent discussion in the medical field concerning the potential for “myokines” in improve metabolic stability and enhance longevity. We have found that specific pulsed magnetic fields stimulate many of the same cellular pathways as exercise and accordingly activate the production of many accepted myokines in vitro. The potential hence exists for PEMF-based therapeutic therapies to condition the body with healthful myokines, recapitulating the healthful benefits of exercise. Indeed, our initial animal studies have shown this possibility to be realized. What remains to be shown is just how closely the parallels exist. This project entails the further optimization of these fields to stimulate myokine expression for ultimate translation into clinical studies at the NUS/NUH.

Department of Obstetrics & Gynaecology Prof Yong Eu Leong [email protected] Dr Susan LOGAN [email protected]

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Integrated Women’s Health Program (IWHP) Singapore is globally one of the most rapidly aging societies. With improvements in health care, life expectancy in females has increased from 68 years to 85 years over the past 45 years. In 2013, women aged >45 years comprised 40% of Singapore’s female Page 70

population &, as such, would expect to spend half of their lives in the menopausal transition & menopause. Many apparently well midlife Singaporean women attend gynecology for routine health screens and care. Currently this care is specialty centric but with a recognition that this age group face multiple health issues including problems relating to menopausal symptoms, bone health and osteoporosis, sexual health, cancer, cardiovascular disease, emotional health, mobility, balance & muscle strength. Many are unrecognized & unmet and all directly influence each other. Improving health care delivery and outcomes depends on systematically establishing the prevalence of health conditions and ultimately health-care needs. In addition, previous research has primarily focused on men. Gynaecologists are ideally placed to bring the focus to women & introduce individualized and innovative healthcare solutions for the prevention and timely treatment of chronic conditions such as hip fractures, cardiovascular disease/stroke, dementia, cancer, and emotional and healthy relationships. The IWHP consists of five elements: 1. A cross sectional study to assess the prevalence & correlates of health conditions and risk factors across multiple systems (cardiovascular, mental, musculo-skeletal, sexual and gynaecological) among a clinical population of women ages 45-69 years in Singapore. The assessment tool includes a questionnaire (electronic in 9/2015), biophysical measurements, blood sampling, short physical performance battery & whole body (bone, muscle, fat) DXA. a. To use this cross sectional information to compare the prevalence of health conditions in the IWHP to that of previously published findings from samples of women living in the United States & globally b. To use the fasted blood samples to explore novel biomarkers in relation to disease prediction 2. To integrate research findings within a treatment setting. Participants will be provided results from their research assessments and will be given referrals to internal resources. This model of care will be termed the Integrated Women’s Health Program (IWHP). 3. Using the IWHP as scaffolding for recruitment of eligible patients, it is proposed to initiate interventional clinical trials after identifying specific needs 4. To undertake a longitudinal assessment of this mid-life female cohort Category: Synthetic Biology Department of Biochemistry Principal Investigator

Project Title with a brief description

A/Prof Matthew Chang [email protected]

Engineering microbes for therapeutic applications Synthetic biology aims to engineer genetically modified biological systems that perform novel functions that do not exist in nature, with

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reusable, standard interchangeable biological parts. The use of these standard biological parts enables the exploitation of common engineering principles such as standardization, decoupling, and abstraction for synthetic biology. With this engineering framework in place, synthetic biology has the potential to make the construction of novel biological systems a predictable, reliable, and systematic process. While the development of most biological systems remains largely ad hoc, recent efforts to implement an engineering framework in synthetic biology have provided long-awaited evidences that engineering principles can facilitate the construction of novel biological systems. In our previous study, we engineered probiotic microbes to effectively sense the presence of a human pathogen and kill the pathogen by releasing antimicrobial peptide, as a proof-ofconcept of a novel microbe-based antimicrobial strategy. Our recent efforts are focused on developing therapeutic microbes with programmable and clinically relevant functionalities to fight against human pathogen. To this end, we have implemented functionalities to include the ability of probiotic microbes to swim toward a human pathogen upon sensing and disrupt biofilm, and exert strong antimicrobial activity against the pathogen. This development suggests the possibility that probiotics could potentially be engineered for prevention and treatment of infectious diseases, which may provide an antimicrobial strategy that is complementary to current antibiotic therapies. With this foundational therapeutic cell engineering in place, our aim is focused on engineering microbes to be further endowed with therapeutic functions toward other gutrelated pathogens and conditions and other human disease such as cancer and metabolic disorders. Engineering microbes to produce valuable biochemicals Metabolic engineering of microbes has been applied to synthesize therapeutic chemicals, such as analgesics, antibiotics, anti-malarials, anti-arrhythmics, parasympatholytics, antioxidants and anti-cancer agents. Notably, synthetic biology strategies have shown great potential in biosynthesis of value-added nutraceutical chemicals at low cost. In our previous study, we have developed synthetic microorganisms that produce valuable biochemicals, and show superior tolerance against toxic chemicals (products and byproducts). Additionally, we have also developed AND-gate dynamic controllers of gene transcription and synthetic promoters for metabolite sensing and regulation, i.e. promoters that combine inducible functionality with regulation of protein expression levels upon detection of key intermediates towards the engineering and optimization of bio-catalytic yeast cells. Our recent project with international collaboration on the synthetic yeast genome project (Synthetic Yeast 2.0, http://syntheticyeast.org) would provide extensive insights to explore the synthetic yeast for efficient production of valuable nutraceuticals. By reconstructing various metabolic pathways, we aim to engineer microorganisms (e.g. bacteria, yeast) to synthesize value-added nutraceuticals. Multiple approaches, such as synthetic biology and omics (functional genomics, transcriptomics, proteomics and metabolomics), will be Updated in Jan 2018

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integrated for pathway reconstruction and flux-balance, analyses of targets (genes, transcripts, and metabolites), and tolerance improvement. Successful delivery of this project could extensively expand our knowledge on value-added chemical biosynthesis pathways in microorganism, and potentially lead to industrialization and commercialization. Developing synthetic genome and chromosomes in microbes The International Synthetic Yeast Genome Project (Sc2.0) has been recently formed to complete the design, synthesis and assembly of the genome of Saccharomyces cerevisiae, which will accomplish the construction of the first ersatz eukaryotic genome. The organism was chosen because its genome is relatively compact and well understood. One underpinning hypothesis of this project is that the “synthetic yeast” approach offers a customised platform that enables us to understand fundamental questions pertinent to characteristics of genomes such as genome organisation, gene structure, gene regulations, functions of RNA splicing and evolution. Yeast chromosome will be re-designed and constructed to fully assemble synthetic chromosome. We envision that using synthetic chromosomes and genome, we will be able to answer profound questions about the properties of yeast metabolism and cellular machinery. A/Prof Yew Wen Shan [email protected]

Current Synthetic Enzymology research interest focuses on protein engineering and biocatalysis, with emphasis on using structural and mechanistic enzymological knowledge to develop therapeutics. We are involved in the following research areas: 1. Defining Synthetic Enzymology as an enabling platform for the

2.

3. 4. 5. 6.

Dr Chester Drum [email protected]

Construction of Next-Generation Synthetic Biology solutions for Pharma and Consumer Businesses. Using Synthetic Enzymology to discover and develop novel therapeutic biomolecules, in the areas of infectious diseases, metabolic disorders, cancer and aging. Developing anti-virulence Quorum-Based Technology for use in the biomedical industry. Using Synthetic Lixiviant Enzymology for Biomining of Electronic Wastes for Environmental Sustainability. Development of Lead Compounds Targeting Enzymes involved in Metabolic Cancer. Using Synthetic Enzymology for the Construction of Therapeutic (Probiotic) Cells for the Treatment of Metabolic Diseases and Infectious Diseases.

A Protein Dynamic Approach to Engineered Translational Therapeutics “The protein folding problem” is one of the most interesting in biology --how does a linear peptide with more conformations than there are atoms in universe, know to fold into essentially a single functional

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conformation? Can biochemical engineering fundamentally effect this process in a way to make it amenable to oncological or cardiovascular treatments? In this project you will use a novel form of protein expression invented by our lab to study protein folding dynamics and novel cellular uptake mechanisms for drug delivery. The project is a continuation of research begun at Massachusetts Institute of Technology and will create nanometer scaled uptake vesicles for the delivery of active protein substrates. Our lab has a full time synthetic chemist, biochemist and materials engineer in addition to full molecular biology and protein biosynthesis support which is an excellent setting for a graduate student who wants an efficient approach to a high impact problem, not bounded by a single technique. Although Prof. Drum was clinically trained as a cardiologist (BWH, Harvard Med School) he is also an award winning structural biologist who is interested in developing talented students into future scientific leaders. You are invited to enquire if interested. Photon mediated therapeutic modulators Electromagnetic radiation is an appealing choice for modulation of biological processes within a living organism. From the historical treatment of cancers through X-ray therapy to the recent advent of optogenetics, the ability of light to permeate living tissues without creating off target effects is well proven. Converting incident radiation to physical work or chemical activity, however, presents a major continuing challenge. Tissue and, in particular blood, have wide absorption ranges and permit relatively narrow bandwidths of electromagnetic radiation to penetrate beneath the skin surface. This project will use a chemical biology approach to implement novel, highly photoisomerizable small molecule sensitizers to convert relevant electromagnetic radiation into cell and protein-specific biological actions. Our major work in this area is currently being prepared for publication and will have many follow-on studies of equally high impact. Disease areas amenable to this approach include cardiovascular, cancer and ophthalmic applications. Our lab has a full time Ph.D. – level chemist, biochemist and spectroscopist to aid a motivated graduate student in project design and completion. If interested, please contact the PI for discussion as the ultimate experimental design will tightly incorporate the intellectual goals of the student in addition to leveraging the current results within the lab. Category: Other Diseases Department of Anatomy Principal Investigator

Project Title with a brief description

Dr Polly Leilei CHEN [email protected] [email protected]

Understanding transcriptome landscape of the human developing liver and the role of transcriptome changes during liver differentiation and development During the natural growth of the human fetal liver, the transcriptome

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actively changes following different developmental stages. We hypothesize that by investigating the patterns of change that occur in the human fetal liver transcriptome, regulatory networks required for development and differentiation of the liver can be elucidated. Using fetal liver specimens of 14, 18 and 22 weeks of gestation, we are interested in studying two major processes that contribute to transcriptome diversity, namely RNA editing and alternative splicing as well as identifying anti-sense transcriptome during liver development. In this project, RNA-sequencing was performed on said specimens, and we intend to screen for candidate genes/events based on 1) magnitude of change between various gestational groups and 2) biological significance. We will then investigate their functional roles using in culture-based cell models, then proceed to small animal models to study in vivo effects. We have successfully established cell models simulating the process of differentiation from human embryonic stem cells (hESCs) to mature hepatocytes. Knowing the regulatory mechanisms of liver differentiation and proliferation may aid in progress of chronic liver treatment, where native liver cells of patients may be coaxed into proliferation under the influence of appropriate molecular signals. New insights gained on how hepatic progenitors may be instructed to proliferate or differentiate and can be applied to develop in vitro hepatocyte differentiation and maturation protocols, ultimately contributing to populate bio-artificial liver assisted devices for liver dialysis, or facilitate toxicology studies in the pharmaceutical industry.

Department of Biochemistry Principal Investigator

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Project Title with a brief description

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Dr Long Yun Chau [email protected]

Nutrient Signaling and Metabolism The role of energy sensing network in exercise-induced skeletal muscle adaptation The beneficial effects of exercise on health are widely recognized. Exercise could improve and prevent chronic metabolic disorders such as diabetes and obesity. However, the cellular mechanism behind these effects is largely unknown. Optimization of exercise regimen or pharmacological recapitulation of exercise effects requires close examination of intramuscular events that triggers the exercise adaptations. Our previous and current work has aimed to define the mechanism that drives exercise-induced metabolic adaptation in skeletal muscle. Exercise induces a multitude of cellular disturbances in muscle including decreased intramuscular energy levels. Exercise-induced energy deficit is considered an important stimulus for the beneficial metabolic outcomes. AMP-activated protein kinase (AMPK) is an important signal transducer which is activated in response to energy stress during exercise. Our previous work in mouse models provided evidence that activation of this protein kinase is sufficient to increase skeletal muscle glycogen store, fatty acid utilization and muscle endurance – benefits that are derived from exercise training. Conversely, disruption of muscle AMPK signaling accelerated the progression of muscle fatigue and impaired exercise-mediated metabolic gene expression. These results provided important evidence that AMPK signaling mediates some of the exercisemediated metabolic responses in skeletal muscle. In the current study, we hypothesize that AMPK plays a role in autophagy and amino acids metabolism in skeletal muscle, and this effect is critical for metabolic benefits of exercise and endurance. Role of insulin-induced glycolysis in the regulation of PGC1αmediated myotube lipid metabolism. Our body is subjected to irregular nutrient supply, including the transition between fasting and feeding. Therefore, the ability of our body to selectively store and utilize different energy substrates is critical for energy balance. Given the substantial mass and energy consumption, skeletal muscle plays a critical role in the regulation of energy balance, and impaired skeletal muscle metabolism is closely associated with metabolic diseases such as type 2 diabetes and obesity. Thus, investigation of skeletal muscle energy substrate metabolism is critical for the understanding of pathophysiology of metabolic diseases. Under postprandial conditions, insulin stimulates skeletal muscle glucose uptake and utilization via Akt, a protein kinase that activates downstream effectors of insulin. Conversely, skeletal muscle increases lipid oxidation under fasting conditions, which coincident with the induction of PGC1α - a transcription coactivator that has been implicated in the induction of lipid metabolic gene program. Nonetheless, the interaction between the disparate insulin and PGC1α pathways which are activated under contrasting nutritional states remains largely unknown. This research

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project aims to establish the role of nutrient (glucose) and hormone (insulin) in the regulation of PGC1α cultured myotubes, and the impact of this regulation on lipid metabolic gene program in myotubes. Role of insulin-like signaling in the regulation of SIRT1 deacetylase The loss of skeletal muscle mass and metabolic function is a common feature of muscle pathologies associated with inactivity (atrophy), aging (sarcopenia) and diseases (cachexia). The loss of muscle function is mainly attributed to elevated protein degradation and resistance to anabolic effects of growth hormones such as insulin and insulin-like growth factor 1 (IGF1). These anabolic hormones are critical signals that stimulate skeletal muscle glycogen and protein synthesis (anabolic metabolism), predominantly via the Akt kinase pathway. Conversely, nutrient deprivation activates Silent Information Regulator T1 (SIRT1) deacetylase and forkhead box protein O1 (FOXO1) proteins that induce catabolic metabolism in skeletal muscle. The increase of catabolic over anabolic pathway may be a molecular basis for the loss of muscle mass and function. Nonetheless, the mechanism by which such delicate balance is regulated remains largely unknown. This proposal aims to evaluate the novel role of insulin-like signals in the regulation of SIRT1, and the unknown function of SIRT1 in the regulation of glucose and amino acid metabolism in cultured myotubes. The specific aims are (1) to determine if insulin-like signals regulate SIRT1 via nutrient metabolism, (2) to establish the role Akt in the regulation of SIRT1induced FOXO1 pathways, and (3) to examine the impact of AKTSIRT1 pathways on energy substrate metabolism. Cultured myotubes will be treated with insulin or IGF1 in the absence of presence of metabolic inhibitors to determine the function of these hormones on SIRT1 function. The role of Akt in the regulation of SIRT1-dependent FOXO1 activity will be evaluated via pharmacological and genetic alteration of the kinase and deacetylase. The metabolic effects of Akt-SIRT1 pathway will be determined by direct measurements of glucose and amino acids metabolism in myotubes to provide direct readout for changes in metabolic fluxes. The research will provide important insight into the pathophysiology of skeletal muscle metabolism, via the identification of insulin-like hormones as regulators of SIRT1. It will also provide critical evidence for SIRT1 and IGF1 as potential drug targets for muscle wasting diseases. Metabolic and Oncogenic Signaling The role of amino acids in growth factor signaling FGF19 is a hormone-like FGF that is released by the small intestine in response to a rise in bile acid levels during the post-prandial state. FGF19 plays a critical role in cancer as well as metabolic signaling. Upon its release, FGF19 will travel to the liver via the hepatic portal vein. It is known that FGF19 activates hepatic anabolic metabolism Updated in Jan 2018

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(biosynthesis) which is required for cell growth, but the precise mechanism of this regulation remains poorly understood. Although previous studies have shown that FGF19 activates components of the protein translational machinery through the mitogen-activated protein kinase (MAPK) signaling pathway, the role of mechanistic target of rapamycin complex 1 (mTORC1) in FGF19 action remains unclear. This study will characterize whether FGF19 also mediates its effect through mTORC1 via activation of its downstream target P70S6K. This project will evaluate the contribution of canonical ERK/P90RSK or the PI3K/AKT pathways in the regulation of FGF19 action. Although growth factors are known to activate cell growth, this project will characterize the impact of amino acids on FGF19 action. We will address the fundamental question in biology – what kind of signaling will a growth factor induce in the absence or presence of nutrient availability? How does nutrient such as amino acid or glucose modulate the action of growth factor?

Department of Medicine Principal Investigator

Project Title with a brief description

Dr Chester Drum [email protected]

Mass spectroscopic biomarker discovery for individualized prediction of pharmacokinetics and disease. The way in which different ethnicities and individuals metabolize drugs can be represented as an intricate web of biochemical changes to prescribed medicines. When this variance in pharmacokinetic metabolism is cast against an electronic medical record database that documents both adverse drug reactions and beneficial effects, the total data set can be treated as a discovery mechanism for unexpected benefits and problems relevant to commonly prescribed medicines. In this project you will interface with a dedicated triple quadrapole tandem mass spectrometer to develop novel detection methods for interesting drug metabolites and use this information to dissect exposure dependent effects of the measured drugs and their ability to effect markers of disease. Using established, large market medicines in new ways and understanding hidden risks and benefits to the Asian market will be critical to healthcare in the upcoming decades. This project seeks to position the graduate candidate to take advantage of this rapidly expanding scientific and regulatory field and develop skills relevant to the implementation of large- market pharmaceutical safety and effectiveness strategies. Because this project can be relevant to both cardiovascular and cancer therapies, the ultimate experimental design will tightly incorporate the intellectual goals of the student in addition to leveraging opportunities already established within the

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lab. Bioinformatics of genetically guided precision medicine Natural variation in the way patients respond to pharmaceuticals is large and even more pronounced amongst Asian ethnicities. Physicians often agonize over the appropriateness of a specific dose of a drug for a given patient. Yet, for that same dose, individual patients may experience up to 20 fold differences in their exposure to the drug due to biological variance in pharmacokinetic biology. The unpredictability of this variance is a major impediment to the future of precision medicine. In this project, you will combine simple pharmacokinetic modeling with an electronic medical record and sequence information from both specific genetic variants and next generation sequencing approaches. Patient-specific pharmacokinetic phenotypes will be provided by the quantification of parent drug and its complete set of metabolites in each patient using a dedicated mass spectroscopy unit within the lab. The ultimate goal of the project is to invent new bioinformatic algorithms for the genetic prediction of drug response and metabolism. The ability to interpret large datasets of patient information and drug metabolism data is a wide open opportunity and has major implications for the understanding of fundamental problems in both pharmaceutical and basic science. Department of Orthopaedic Surgery Principal Investigator

Project Title with a brief description

Prof James Hui [email protected]

Effect of electromagnetic cues on stem cell behavior in conductive constructs Stem cell fate is influenced by its microenvironment. Manipulation of extracellular biophysical and/or biochemical cues can improve the efficiency of tissue engineering approaches using stem cells. Physical cues at the cell-matrix interface, including the matrix mechanical, geometrical and topographical properties can trigger profound cellular response. In addition, dynamic stimulation, in the form of mechanical and electric/magnetic stimulation, is also critical factors in stem cell proliferation and differentiation. In Stem Cell Biology and Cartilage Research Group (NUS Tissue Engineering Program), our focus is on the improvement of the efficacy of stem cell therapy in cartilage regeneration. We have identified specific biochemical, topographical cues and mechanical stimulation regiment for the directed differentiation of stem cells to specific cartilage phenotype. In this project, the effect of electromagnetic field on human mesenchymal stem cell (MSC) differentiation to cartilage cells will be investigated in a conductive constructs. Conductive polymers with specific electrical properties on material surfaces, have been used for nerve, cardiac and bone tissue engineering. The project will explore MSC response to electromagnetic field in different 2D and 3D conductive constructs with variable parameters by

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manipulating the physical cues of the cell-matrix interface. The study will include elucidation of the mechanotransduction mechanisms that affect specific functional outcomes. Student will leverage skills in scaffold fabrication (including macroporous and nano-fibrous) and surface modification, scaffold characterisation; cell culture, histology, optical microscopy and, molecular biology. References: • Raghothaman D, Leong MF, Lim TC, Toh JKC, Wan ACA, Yang Z, Lee EH. Engineering cell matrix interactions in assembled polyelectrolyte fiber hydrogels for mesenchymal stem cell chondrogenesis. Biomaterials 2014;35(9):2607-16. • Wu Y, Law JBK, He AY, Low HY, Hui JHP, Lim CT, Yang Z, Lee EH. Substrate topography determines the fate of chondrogenesis from human mesenchymal stem cells resulting in specific cartilage phenotype formation. Nanomedicine. 2014 10(7):150716. • Zhang T, Wen F, Wu Y, Goh GSH; Ge Z, Tan LP, Hui JH, Yang Z. Cross-talk between TGF-beta/SMAD and integrin signaling pathways in regulating hypertrophy of mesenchymal stem cell chondrogenesis under deferral dynamic compression. Biomaterials 2015;38:72-85. Prof Lee Eng Hin [email protected]

Application of biophysically sorted chondrocytes in articular cartilage regeneration In Stem Cell Biology and Cartilage Research Group (NUS Tissue Engineering Program), our focus is on the improvement of the efficacy of cell therapy in cartilage regeneration. Articular cartilage is a weight-loading tissue in which the biomechanical properties of the tissue are dependent on the stratified structural organisation of the tissue. Different depth-related zones of cartilage vary greatly in terms of cellular density, morphology and metabolic activity, as well as the composition and structural arrangement of the extracellular matrix components. These zonal biochemical differences in turn lead to significant variations in strains and stresses experienced by the cells in different zones during joint loading. The approach to engineer a fully functional tissue based on zonal chondrocytes for treatment of cartilage defeat to date presents many difficulties. Apart from the limited amount of cells that can be harvested from each zone, the lack of reliable and easy-to-handle zonal cell sorting protocols severely hinders the practicality. In this project, in collaboration with Singapore-MIT Alliance for Research and Technology (SMART) Centre, we will apply inertial microfluidics to separate different zonal chondrocytes from cartilage tissue, as well as from stem cell-derived tissue, based on the size-variation of zonal chondrocytes. The project involves the optimisation of the separation protocols, characterisation of the separated chondrocyte subpopulations, and validation of the efficacy of the zonal chondrocytes to restore hierarchically-organized cartilage in an animal cartilage defect model. Student will leverage skills in microfluidics, mechanics, cell culture, histology, molecular biology, optical microscopy and in vivo

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experiments. •

•

•

•

A/Prof Wilson Wang [email protected]

Raghothaman D, Leong MF, Lim TC, Toh JKC, Wan ACA, Yang Z, Lee EH. Engineering cell matrix interactions in assembled polyelectrolyte fiber hydrogels for mesenchymal stem cell chondrogenesis. Biomaterials 2014;35(9):2607-16. Wu Y, Law JBK, He AY, Low HY, Hui JHP, Lim CT, Yang Z, Lee EH. Substrate topography determines the fate of chondrogenesis from human mesenchymal stem cells resulting in specific cartilage phenotype formation. Nanomedicine. 2014 10(7):150716. Poon Z, Lee WC, Guan G, Nyan LM, Lim CT, Han J, Van Vliet KJ. Bone marrow regeneration promoted by biophysically sorted osteoprogenitors from mesenchymal stromal cells. Stem Cells Transl Med. 2015 Jan;4(1):56-65. Lee WC, Shi H, Poon Z, Nyan LM, Kaushik T, Shivashankar GV, Chan JK, Lim CT, Han J, Van Vliet KJ. Multivariate biophysical markers predictive of mesenchymal stromal cell multipotency. Proc Natl Acad Sci U S A. 2014 Oct 21;111(42):E4409-18.

Evaluation of Magnetic Resonance Image (MRI)-Based Novel 3Dimensional (3-D) Printed Scaffold Meniscus to Replicate and Replace Native Meniscus in an Ovine Model: In Vivo Studies of Morphology, Function, Viability and Response The knee menisci are fibro-cartilaginous crescent-shaped wedges that are essential to knee joints in load bearing, shock absorption and joint stability. Their morphology and internal structure are crucial to their normal functioning. Meniscal injuries are common in both younger individuals (sports injuries) as well as older patients (degenerative tears), and increase the risk of post-traumatic knee osteoarthritis. Current treatment modalities (meniscectomy, repair) are often suboptimal due to poor healing potential and disruption of normal structure, while allograft transplantation is limited by mismatch and availability issues. This clinical need has prompted research in new scaffold technologies for meniscal replacement, but current techniques do not recreate the internal microstructure crucial to meniscal function. Our group has developed a novel patented 3-D printing process using drop-on-demand micro-dispensing technology that can create multimaterial meniscal scaffolds with morphology and microstructural patterns similar to native menisci. We believe this specific advantage in being able to replicate internal microstructural orientations is a breakthrough in bioscaffold technology that will enhance the chances of long-term bio-implant success. Our 3-D printed cell-laden polymer/collagen meniscal scaffolds have been characterized in vitro for material and biomechanical properties and biological responses, with promising results. The proposed present investigation aims to develop this technology further, using an in vivo ovine model; we aim to validate the customized 3-D replication of meniscal morphology by MRI-based scaffold printing in

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a large animal model. The meniscal scaffolds (without and with autogenous cell seeding) will be surgically implanted in the sheep knees, and assays performed for knee function and complications, biocompatibility and inflammatory response (with sham surgery and meniscectomy controls). After euthanasia and knee harvesting at time intervals, further evaluations will be made for biomechanical function and histological, cellular and molecular changes. This project aims to support the downstream development of customized meniscal implants in clinical applications. Prof Wong Hee Kit [email protected]

Bone regeneration using NELL-1, a non-BMP2 osteoinductive growth factor, delivered by novel carriers in a lumbar spinal fusion model Bone regeneration in spine is a common technique used to treat trauma, low back pain and deformities. With the aging population, the number of spinal fusion surgeries performed annually is increasing significantly, which adds to the health care burden globally. The use of autologous bone graft, gold standard for spinal fusion, was limited by associated complications such as donor site pain and unsatisfactory fusion rate, with reported failure of fusion of up to 45% Osteobiologics were introduced as substitutes for autologous bone graft for bone regeneration. Bone Morphogenetic Protein-2 (BMP-2) is one of most potent bone growth factor approved by FDA for human spinal fusion procedure. However, safety concerns from complications associated with BMP-2 usage were raised recently. There is still an unmet need to develop a safe and efficient bone regeneration device. Due to the adverse effects reported recently that are associated with the BMP-2 usage in clinical situation, there is a need to substitute BMP-2 with new and more specific bone regeneration growth factors such as NELL-1. NELL-1, one of the new osteobiologics under investigation, is known to be more specific in bone regeneration than BMP-29. It has been shown that it could induce bone formation in vivo models from rodents to large animals without significant complications. We hypothesize that localization and controlled release of a nonBMP-2 based growth factor, NELL-1 in one or both of our two growth factor carriers, PEC and BNPP, will improve the efficiency and efficacy of bone regeneration in spinal fusion. This effect will be evaluated in small and large animal lumbar interbody fusion models. If successful, this study will introduce a new generation of non-BMP based bone graft substitute consisting of NELL-1 matched with an optimal carrier that will significantly improve the clinical outcome of spinal fusion procedure.

Department of Otolaryngology Principal Investigator

Project Title with a brief description

A/Prof Susan Rickard Liow [email protected]

Cognitive-linguistic Screening for Early Stage Dementia (Mild Neurocognitive Disorders)

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There are many different types of dementia and reliable differential diagnosis in the early stages remains a challenge even with advances in neuroscience. This is because the behavioral pencil and paper tasks available for screening people with suspected onset of cognitive-linguistic deficits are not sufficiently sensitive, and most focus on memory problems. One variant of Frontal-Temporal Dementia (FTD) is characterized by language processing problems (spelling, writing, naming) in relatively young adults (