Offered by The University of Melbourne Department of Paediatrics and The Murdoch Childrens Research Institute
To find out more about our course and our projects please come to our information night on:
Thursday 16 September 2010
4.00pm – 6.00pm
Ella Latham Theatre, Royal Children’s Hospital
Please note that the project list will be updated regularly with new projects and opportunities.
- Understanding age-related differences in the structure of coagulation proteins and their interaction with anticoagulants.
- The role of Epigenetics in Paediatric Leukaemia development and outcome
- Using Next Generation Sequencing to Discover Novel Genes that cause Mitochondrial Disease.
- Understanding basal ganglia development in preterm infants using magnetic resonance imaging
- Understanding neurodevelopmental outcomes of late preterm neonates using magnetic resonance imaging
- Molecular epidemiology of Streptococcus pneumoniae
- Understanding motor development in the very preterm child using diffusion magnetic resonance imaging
- The effects of prematurity on the visual system of children born preterm
- Understanding brain development of extremely preterm adolescents using diffusion magnetic resonance imaging
- 1. Epigenetic variation in newborn twins: effect of maternal diet and environment and underlying genetic make up
- Investigating the utility of epigenetic analysis of placenta-derived DNA for predicting adverse pregnancy outcomes.
- Neuropsychological functioning in children with type 1 diabetes
- Neuropathogenic mechanisms of mitochondrial dysfunction
- Cardiac Molecular signaling mechanisms during the progression of heart failure
- Cardiac molecular signaling mechanisms in survival adaptation to hypoxia and post-operative stress recovery
- Determining cord blood stem cells with cardiac fate for the repair of congenital myocardial dysfunction
- DNA methylation of mobile genetic elements in germ cells
- Small RNAs and the final stage of sperm development
- The Effect of Oral Immunotherapy on Immunological Markers of Peanut Allergy
- Immunological effects following prenatal probiotic supplementation for the prevention of eczema
- Immunological effects following administration of prebiotic supplemented hydrolysed formula for the prevention of eczema
- Why does this mouse have funny bones?
- Investigating the effects of increased IL-11 expression in the stomach
- Tumour suppressor genes in gastric cancer
- Identification of Rotavirus vaccine strains in children with gastroenteritis.
- Understanding the role of infectious agents in children with early onset Crohn’s disease
- Childhood Arthritis – the role of epigenetics in determining risk of disease
- The role of epigenetics in the regulation of body weight regulation and metabolism in childhood obesity.
- Analysis of mouse models of human birth defects
- Large-scale screen of genes controlling skeletal development
- Analysis of mouse models of human birth defects
- Large-scale screen of genes controlling skeletal development
- Lung function genes and airway remodelling in asthma.
- Epithelial remodelling and repair in airway disease.
- The molecular signalling pathways that cause osteoarthritis
- The contribution of protein misfolding (“unfolded protein response”) to inherited cartilage and bone disease
- Identifying TRPV4 interacting partners critical in human disease phenotypes
- TRPV4 and skeletal development
- Collagen VI and WARP interactions during neurological development
- Stomach inflammation and epigenetic silencing of the TFF2 tumour suppressor gene
- Epigenetic factors in Triplet-Repeat Associated Diseases
- Modifying and Preventing Heart Disease in Mice with Mitochondrial Cardiomyopathy
| Dr Vera Ignjatovic Haematology Research Critical Care and Neurosciences T 99366520 E verai@unimelb.edu.au |
Professor Paul Monagle Haematology Research Critical Care and Neurosciences T 93455161 E paul.monagle@rch.org.au |
Haemostatic system of children evolves with age, with marked physiological differences in the concentration of most haemostatic proteins, a concept known as Developmental Haemostasis. We have previously observed that these quantitative differences in haemostatic proteins do not explain all of the age-related differences in the effect of anticoagulants, suggesting the role for qualitative differences. Studies performed in our laboratory confirmed that fibrinogen isolated from neonatal and child plasma is qualitatively different to that isolated from adult plasma. In addition, we have demonstrated clinically significant age-related differences in the anticoagulation effect of the anticoagulant Heparin, hypothesized to be due to the age-specific differences in binding of this drug to coagulation as well as other plasma proteins. Using state of the art proteomic methodology, this proposal will build on our current and previous work and contribute significantly towards developing treatment strategies for children based on sound experimental evidence. This proposal will be the first attempt to: 1: Determine the extent of age-related differences in structure and binding kinetics of key coagulation proteins in humans (Fibrinogen, Antithrombin, and Thrombin); 2: Investigate the age-related differences in the interaction of haemostatic proteins with clinically relevant anticoagulants.
2. The role of Epigenetics in Paediatric Leukaemia development and outcome
| Dr Nicholas Wong Developmental Epigenetics Early Development and Disease T 8341 6205 E nick.wong@mcri.edu.au |
Dr Jeffrey Craig Developmental Epigenetics Early Development and Disease T 8341 6346 E jeff.craig@mcri.edu.au |
| Dr Richard Saffery T 8341 6341 E |
Leukaemia is the most common form of cancer in children, accounting for over 30% of newly diagnosed cases. Most cases involved specific genomic rearrangements (translocations). However, these are neither sufficient nor absolutely necessary for disease development. Despite the fact that ~80% of cases are successfully treated by chemotherapy, the underlying causes of childhood leukaemia remain unclear and cannot be explained by genetic or environmental factors alone. Epigenetics is an emerging field examining the modulation of gene expression in the absence of underlying genetic change. We believe disruption of epigenetic profile could play a major role in the aetiology of paediatric leukaemia in conjunction chromosome translocations. This project will catalogue epigenetic changes at gene promoters from archived matched leukaemia and remission bone marrow samples in an attempt to identify changes in associated with development or outcome of specific leukaemia subtypes.
3. Using Next Generation Sequencing to Discover Novel Genes that cause Mitochondrial Disease.
| A/Professor David Thorburn Mitochondrial Research Laboratory and Community Genetics T 83416235 E david.thorburn@mcri.edu.au |
Dr Alison Compton Mitochondrial Research Laboratory and Community Genetics T 6287 E alison.compton@mcri.edu.au |
Mitochondria are the powerhouses of the cell, generating cellular energy through the oxidative phosphorylation (OXPHOS) system. Pathogenic mutations in genes required for correct assembly of the OXPHOS protein complexes result in a variety of neurodegenerative disorders collectively known as mitochondrial diseases. Nearly 100 (nuclear and mitochondrial) genes are known causes of mitochondrial disease, however ~50% of patients still do not have a molecular diagnosis with many more novel disease genes awaiting discovery. Recently, we used a pooled high-throughput sequencing strategy to screen 103 genes encoding complex I subunits and proteins putatively involved in complex I biogenesis in a cohort of 103 complex I deficient patients. Our patient cohort included 43 patients with known mutations, allowing us to demonstrate a sensitivity of 83% for detecting nuclear DNA mutations. We identified a new genetic diagnosis in 22% of the remaining 60 patients, including mutations in known subunit and complex I assembly factor genes. Most excitingly we also identified mutations in two novel complex I disease genes, confirming their pathogenicity by performing lentiviral correction studies in patient fibroblasts. This project successfully acted as a pilot ‘proof of principle’ study for our current larger project in which we have used hybrid selection enrichment for 1400 mitochondrial genes combined with massively parallel targeted sequencing in a heterogeneous group of 50 patients with biochemically defined OXPHOS deficiency. This project will follow up on several of the many sequence variants identified to discover novel OXPHOS disease genes and determine their normal function and disease pathogenesis using a combination of cell biology, molecular biology and biochemical approaches.
4. Understanding basal ganglia development in preterm infants using magnetic resonance imaging
| Dr Deanne Thompson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E deanne.thompson@mcri.edu.au |
Dr Peter Anderson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E peter.anderson@mcri.edu.au |
| Dr Rod Hunt T 93456265 E |
A/Professor Gary Egan T 83441938 E |
Up to 50% of preterm infants born <30 weeks gestational age or <1250g birth weight display cognitive and motor deficits. Considering the involvement of the basal ganglia in learning and motor control, this structure is of particular interest in preterm infants. Structural magnetic resonance imaging enables examination of the basal ganglia in-vivo. The aims of this project will be to compare 3D volumes of the various basal nuceli of preterm infants with full-term controls at term equivalent age, and to determine the perinatal causes and functional consequences for alterations to the basal ganglia in this vulnerable population.
| Dr Deanne Thompson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E deanne.thompson@mcri.edu.au |
Ms Jeanie Cheong Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 83453771 E jeanie.cheong@thewomens.org.au |
| Dr Peter Anderson T 93454830 E |
Neurodevelopment of late preterm infants born between 32 to 36 weeks' gestational age has not been extensively studied, even though there is evidence to suggest that such infants have neurobehavioural and educational difficulties in comparison to their term-born peers. The aim of this study (LaPrem) of late preterm infants is to compare quantitative MRI characteristics of late preterm and healthy term-born infants at term equivalent age. Volumetric analyses of brain tissues such as gray and white matter will be performed, as well as segmentation of the frontal, temporal, parietal and occipital lobes, cerebellum and hippocampus. Diffusion MRI will be used to obtain measures of microstructural integrity, and the connectivity of various white matter bundles will be estimated using advanced MRI processing techniques such as tractography. Comprehensive neurobehavioural assessments will also be performed on these infants in order to assess the consequences of late preterm birth.
6. Molecular epidemiology of Streptococcus pneumoniae
| Dr Eileen Dunne International Child Health Infection, Immunity and Environment T 6531 E eileen.dunne@mcri.edu.au |
Dr Catherine Satzke International Child Health Infection, Immunity and Environment T 83416438 E catherine.satzke@mcri.edu.au |
| Professor Kim Mulholland T 93454977 E |
Pneumonia is the leading cause of death of children under the age five. The majority of cases are due to infection by Streptococcus pneumoniae (the pneumococcus), which is commonly carried in the nasopharynx of children. The seven-valent pneumococcal conjugate vaccine (PCV7, Prevnar) protects against seven common invasive serotypes, but its use has been associated with an increase in invasive disease caused by serotypes not included in the vaccine. S. pneumoniae is a very diverse species, with over 90 capsular serotypes. New strains can arise through recombination or mutation, and multidrug resistant clones have become increasingly common worldwide. Factors that contribute to clonal conservation or expansion include selective forces such as the immune response and antibiotic pressure, or the acquisition of genes that enhance bacterial transmission and colonisation. Our laboratory is interested in characterising the molecular epidemiology of S. pneumoniae carriage in populations with high disease burden. Multilocus sequence typing (MLST) provides information on the genetic relatedness between strains by comparing the sequences of seven housekeeping genes. This project will utilise a new mass spectrometry based platform (Sequenom) to perform MLST on S. pneumoniae isolated from children in developing countries. Additional experimental techniques will include antibiotic susceptibility testing and screening for virulence genes to further characterize these strains.
| Dr Deanne Thompson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E deanne.thompson@mcri.edu.au |
Ms Jeanie Cheong Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 83453771 E jeanie.cheong@thewomens.org.au |
| Dr Peter Anderson T 93454830 E |
Up to 50% of preterm infants born <30 weeks gestational age or <1250g birth weight display either severe motor deficits such as cerebral palsy, or milder forms of motor impairment known as developmental coordination disorder (DCD). Tractography is a diffusion MRI processing technique that can provide a virtual 3D representation of the white matter fibre tracts, such as the sensorimotor pathways. The aim of this study is to use tractography to assess the connectivity and microstructural integrity of the corticospinal tracts in 7 year old children born preterm from the Victorian Infant Brain Studies (VIBeS) cohort, compared with healthy term-born children. Perinatal causes and functional consequences for alterations to the corticospinal tracts will also be assessed.
8. The effects of prematurity on the visual system of children born preterm
| Dr Deanne Thompson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E deanne.thompson@mcri.edu.au |
Dr Rod Hunt Neonatal Research Critical Care and Neurosciences T 93456265 E rod.hunt@rch.org.au |
| Dr Peter Anderson T 93454830 E |
One of the many potential difficulties very preterm infants born <30 weeks gestational age or <1250g birth weight face is disruption to visual development. The integrity of the visual pathways is able to be assessed using diffusion MRI. Specifically, tractography is a technique that can provide a 3D virtual representation of the visual tracts, and diffusion measures are able to estimate the axonal integrity and degree of myelination within the white matter tract. The aim of this study is to use diffusion MRI to assess the connectivity and integrity of the visual pathways of 7 year old children born preterm from the Victorian Infant Brain Studies (VIBeS) cohort, compared with healthy term-born children. Perinatal causes and functional consequences for alterations to the visual tracts will also be assessed.
| Dr Deanne Thompson Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 93454830 E deanne.thompson@mcri.edu.au |
Professor Lex Doyle Victorian Infant Brain Studies (VIBeS) Critical Care and Neurosciences T 83453716 E lwd@unimelb.edu.au |
| Ms Jeanie Cheong T 83453771 E |
Dr Peter Anderson T 93454830 E |
The Victorian Infant Collaborative Study (VICS) has been following the neurodevelopment of extremely preterm infants born in 1991-1992. This adolescent cohort is currently undergoing MRI scans. Opportunity exists for a student to investigate various neural networks that are commonly affected by prematurity, using diffusion-weighted neuroimaging techniques such as 3-D white matter tractography. Quantitative diffusion MRI techniques enable insight into the connectivity of white matter fibres, and microstructural integrity of axons and myelin.
| Dr Jeff Craig Developmental Epigenetics Early Development and Disease T 83416346 E jeff.craig@mcri.edu.au |
Dr Richard Saffery Developmental Epigenetics Early Development and Disease T 83416341 E richard.saffery@mcri.edu.au |
Many chronic diseases have an environmental component, and can be influenced even by the environment experienced in the womb. We are searching for the factors that, during pregnancy, influence the epigenetic profile of the fetus and with this, the health of the individual at birth and in later life. Epigenetic factors modify gene structure and function without changing DNA sequence; they are reversible and can be influenced by the environment. We have established a cohort of mothers and their newborn twins and have collected maternal blood and various tissues (including cords, cord blood and placenta) from newborns. Maternal nutrition and other environmental factors will be measured in mothers and are being used in state-of-the-art methods to analyse epigenetic markers in the twins. We are also studying ‘identical’ (monozygotic) twins to assess epigenetic variation at birth independent of genetic variation and to pinpoint the genes associated with low birth weight, which is associated with predisposition to complex disease. Our study also includes fraternal (dizygotic) twins, which can help shed light on the role that dietary/lifestyle and genetic factors have on epigenetic variation of newborns. This project will involve analysis of DNA methylation and/or histone modification and will involve genome-wide microarray analysis. The student will be working in a supportive environment of ten researchers skilled in all the required techniques. Current PhD students have travelled overseas to present their work and published in high impact journals.
| Dr Richard Saffery Developmental Epigenetics Early Development and Disease T 83416341 E richard.saffery@mcri.edu.au |
Dr Jeff Craig Developmental Epigenetics Early Development and Disease T 83416346 E jeff.craig@mcri.edu.au |
| A/Professor Jane Halliday T 83416260 E |
Preeclampsia affects 7-10% of all pregnancies, accounts for 15% of preterm births, and contributes 25% of intrauterine growth restriction. It is directly responsible for 50,000 maternal, and 900,000 perinatal deaths worldwide each year. Inadequate first trimester development of the placenta is now recognised as a primary cause of pregnancy complications such as pre eclampsia (PE) and intra uterine growth restriction (IUGR) with emerging data implicating faulty establishment of epigenetic profile (particularly DNA methylation) in this process. Epigenetics refers to factors associated with genes that switch them 'on' or 'off'. In this study, we aim to define the DNA methylation changes associated with PE in prospectively collected first trimester chorionic villous sampling (CVS) tissue and at birth and to compare this to matched disease-free tissue. This will be done using genome-wide methylation analysis with Illumina 27K BeadChip arrays. Validation of candidate gene methylation differences will be performed by Mass Spectroscopy-based EpiTYPING. Techniques utilised will include DNA isolation from CVS/placental tissue, bisulfite conversion of DNA, PCR amplification, gel electrophoresis, and EpiTyPER analysis. Gene expression levels will also be measured using quantitative RT-PCR. The project will also involve extensive bioinformatic analysis.
12. Neuropsychological functioning in children with type 1 diabetes
| A/Professor Elisabeth Northam Diabetes Early Development and Disease T 83416523 E lis.northam@rch.org.au |
Ms Caroline Nadebaum Child Neuropsychology Critical Care and Neurosciences T 83415634 E caroline.nadebaum@mcri.edu.au |
Type I diabetes mellitus is one of the most common chronic diseases of childhood and adolescence. Affected children and adolescents confront a serious disease burden that includes a daily requirement for exogenous insulin administration, the need to monitor metabolic control and the need to pay constant attention to dietary intake. Complications include blindness, renal failure, premature heart disease, neuropathy and peripheral circulatory failure. Type 1 diabetes may also have a significant impact on the developing central nervous system, as cerebral glucose and insulin levels are frequently abnormal even when diabetes is well controlled. This project forms part of a larger longitudinal study examining neuropsychological outcomes in children with type 1 diabetes. The specific project, suitable for an honours student involves follow-up cognitive assessments of healthy control children.
13. Neuropathogenic mechanisms of mitochondrial dysfunction
| Dr Ann Frazier Mitochondrial Research Laboratory and Community Genetics T 83416287 E ann.frazier@mcri.edu.au |
A/Professor David Thorburn Mitochondrial Research Laboratory and Community Genetics T 83416235 E david.thorburn@mcri.edu.au |
Mitochondrial dysfunction causes a range of early-onset symptoms and contributes to neurodegenerative neurological conditions such as Parkinson Disease. The mechanisms by which mitochondrial disease results in neuronal damage are unknown, therefore the study of these at a cellular level may lead to improved treatment and greater understanding of the role of nuclear- and mitochondrial-DNA mutations in both rare and common conditions. This project will focus on the most common mitochondrial energy production disorder, complex I deficiency, by establishing and characterising cell culture models of complex I deficiency: 1) Neural cell cultures established from two mouse models of complex I deficiency resulting from mutations in two different nuclear encoded complex I subunits. 2) Fibroblast cell lines from patients with nuclear or mtDNA mutations affecting complex I activity, as well as mouse embryonic fibroblasts from our mouse models. We will use a number of techniques in microscopy, biochemistry and molecular biology to study the effects of these mutations, assessing parameters such as the mitochondrial membrane potential, reactive oxygen species, ATP production, apoptosis and cellular calcium dynamics. Such studies could highlight potential therapeutic approaches and allow us to monitor the effects of therapy.
14. Cardiac Molecular signaling mechanisms during the progression of heart failure
| Dr Salvatore Pepe Heart Research Critical Care and Neurosciences T 93454114 E salvatore.pepe@mcri.edu.au |
Dr Freya Sheeran Heart Research Critical Care and Neurosciences T 93454762 E freya.sheeran@mcri.edu.au |
| A/Professor Joe Smolich T 93454571 E |
Congenital and acquired myocardial disorders, despite diverse etiology, commonly involve a reduced capacity to manage oxygen and nitrogen free radical metabolism. Chronic augmented oxidative stress, particularly in fetal and neonatal development, not only leads to post-translational structural modification of proteins, but also impacts gene transcription, ultimately with consequences to structural, metabolic and functional remodeling during adaptive and maladaptive heart failure. These pathological changes remain to be well defined in the developing heart at molecular and cellular level in order for potential therapeutic targets to be identified. Students ideally should have a background in at least one of the following: biochemistry, immunology or pharmacology. Studies will explore novel molecular signaling pathways (intracellular/mitochondrial/nuclear) using unique in vitro and ex vivo models and a range of immunohistochemical, biochemical, molecular, genetic and cell biology methods.
| Dr Salvatore Pepe Heart Research Critical Care and Neurosciences T 93454114 E salvatore.pepe@mcri.edu.au |
Dr Michael Cheung Heart Research Critical Care and Neurosciences T 93455714 E michael.cheung@rch.org.au |
| Professor Igor Konstantinov T 93455200 E |
Dr Freya Sheeran T 93454762 E |
Many congenital heart disorders, involve chronic hypoxic conditions due to one or more cardiovascular structural defects which compromise normal cardiopulmonary blood flow and thus blood reoxygenation. During heart surgery cardioplegic heart arrest and cardiopulmonary bypass impose additional inflammatory and ischemia-reperfusion stress. Ischemic preconditioning (IPC) activates a powerful innate protection via brief intermittent periods of coronary artery ischemia-reperfusion prior to a sustained period of ischemia, thus reducing post-ischemic injury. In animal and human models of ischemia-reperfusion injury, a simple stimulus known as remote ischemic preconditioning (RIPC) has been shown to reduce post-ischemic tissue damage and inflammation. RIPC can be invoked by causing IPC at a site remote from the heart, ie using a pressure cuff to intermittently occlude and reperfuse blood vessels in limbs. In numerous cell types mitochondria have been recognised to be central to IPC where multiple signalling pathways appear to converge to regulate metabolic function. However the molecular and cellular mechanisms that underly the cardioprotection induced by RIPC remain to be defined. Thus, our current goals are to define these intracellular/ mitochondrial/ nuclear signaling pathways using animal models and cell-based studies. New understanding of RIPC will identify specific targets to harness more potent cardioprotective effects in our clinical setting. Students should have a background in one or more of the following: biochemistry, immunology, genetics, physiology, pharmacology.
| Dr Salvatore Pepe Heart Research Critical Care and Neurosciences T 93454114 E salvatore.pepe@mcri.edu.au |
Dr Ngaire Elwood Cord Blood Bank Early Development and Disease T 93456398 E ngaire.elwood@mcri.edu.au |
| Dr Christian Brizard T 93455200 E |
As the heart has recently been found to be capable of self renewal, the current work is aimed at acquiring a basic understanding of the growth and differentiation of cardiac progenitor cells. Cord blood, obtained from the placenta and umbilical cord, contains cells called unrestricted somatic stem cells (USSC) that are capable of forming many different tissues including the heart. Related genetic transcriptional signaling, immune and endocrine regulatory factors that are involved in determining what drives cardiac cell fate will be studied to develop models of myocardial cell recruitment for potential treatment of congenital heart disorders which currently only have palliative surgical treatment options. Beyond cord cell line manipulation and characterization, our goals are projected to facilitate the iterative development of experimental surgical models, working with surgeons and clinicians. Students ideally should have a background in one or more areas such as immunology, genetics, biochemistry or pharmacology, and will predominantly utilise genetic, molecular and cellular techniques.
17. DNA methylation of mobile genetic elements in germ cells
| Dr Jeff Mann Stem Cell Epigenetics Laboratory and Community Genetics T 99366516 E jeff.mann@mcri.edu.au |
Dr Deidre Mattiske Stem Cell Epigenetics Laboratory and Community Genetics T 99366517 E deidre.mattiske@mcri.edu.au |
BACKGROUND: Mobile genetic elements—also called ‘dispersed repetitive transposable elements’ (DRTEs)—make up to one half of the total mammalian genome. Active DRTEs can insert into genes causing mutation and disease. When DRTE insertions occur in the genome of germ cells, the mutations are inherited in the offspring, and can be passed to subsequent generations. All cells use DNA methylation to shut down the promoter activity of DRTEs. However, primordial germ cells (PGCs), which are germ cells at a very early stage, are stripped of DNA methylation as part of a genome-wide reprogramming phenomenon. This leaves them vulnerable to the mutagenic activity of DRTEs. It has been described that one particularly important DRTE, the ‘endogenous retrovirus type II’ (ERVII), is unusual in that it is not stripped of DNA methylation in PGCs. However, these studies have been of the ERVII population as whole, which is comprised mostly of incomplete and inactive ERVIIs. Therefore, in PGCs, the DNA methylation status of complete and potentially active ERVIIs remains unclear. PROJECT AIM: To determine the DNA methylation status in PGCs of a number of selected single complete and potentially active ERVIIs. METHODS TO BE USED: Dissection of mouse embryos and fetuses, flow cytometry to purify germ cells, and DNA methylation analysis of purified germ cells using PCR-based techniques.
18. Small RNAs and the final stage of sperm development
| Dr Jeff Mann Stem Cell Epigenetics Laboratory and Community Genetics T 99366516 E jeff.mann@mcri.edu.au |
Dr Deidre Mattiske Stem Cell Epigenetics Laboratory and Community Genetics T 99366517 E deidre.mattiske@mcri.edu.au |
BACKGROUND: Small non-coding RNAs such as micro RNAs (miRNAs) and small interfering RNAs (siRNAs) produced by the enzyme DICER have a pervasive role in biological processes. We have previously shown that these small RNAs are important for the final stages of oocyte development: DICER-deficient, and hence mi/siRNA-deficient, oocytes fail at ovulation due to a breakdown in chromosome organization. In male germ cells, removal of DICER at an early stage leads to an early failure of spermatogenesis. However, the role of DICER during spermiogenesis is unknown. Spermiogenesis is the final or haploid phase of spermatogenesis, referring to the development of newly formed round haploid spermatids into mature sperm. Investigation of the potential role of DICER in spermiogenesis is of interest as mi/siRNAs could be 1) required for the completion of spermiogenesis, or 2) important for early development, as it has been reported that mi/siRNAs transmit important epigenetic information into the fertilized egg. PROJECT AIM: To examine the role of DICER in mouse spermiogenesis using genetically modified mice that have already been developed in our laboratory. METHODS TO BE USED: Dissection of mouse embryos and fetuses, flow cytometry to purify spermatogenic cells, histology, quantitative real-time PCR.
19. The Effect of Oral Immunotherapy on Immunological Markers of Peanut Allergy
| A/Professor Mimi Tang Allergy and Immune Disorders Infection, Immunity and Environment T 93455911 E mimi.tang@rch.org.au |
Dr Paul Licciardi Allergy and Immune Disorders Infection, Immunity and Environment T 93455554 E paul.licciardi@mcri.edu.au |
Food allergy affects 6-8% of Australian children and 1.5% of children have peanut allergy. Unlike most food allergies that resolve by late childhood, peanut allergy usually persists. Peanut allergy is the most common cause of anaphylaxis in children and the most common cause of death from food anaphylaxis. Currently, there is no cure for food allergy. Management relies on strict avoidance of the food; however, 50% of children have accidental exposure to peanut within 1 year, and 42% of reactions are life-threatening. Anxiety surrounding peanut allergy is extreme due to the rare but devastating possibility of death in a healthy child. A treatment that can induce tolerance would provide a cure. Oral immunotherapy (OIT) is an exciting new approach to induce tolerance in subjects with food allergy. Recent studies suggest that the addition of an adjuvant such as probiotics to OIT can increase the ability for OIT to induce tolerance. In particular, LGG has been shown to have immunomodulatory effects that can promote the development of oral tolerance when administered orally together with antigen. We are conducting a randomised controlled trial (RCT) designed to test the hypothesis that combined administration of a probiotic LGG and peanut oral immunotherapy (P-POIT) is more effective than placebo for the induction of tolerance to peanut in children with peanut allergy. We also hypothesise that P-POIT will 1) improve quality of life and 2) induce immunomodulation with reduced peanut SPT, reduced peanut specific IgE, and increased peanut specific IgG and IgA. This project aims to examine the effects of probiotic peanut OIT on peanut specific IgE, IgG and IgA, and on peanut SPT. The project will involve ELISA techniques.
20. Immunological effects following prenatal probiotic supplementation for the prevention of eczema
| A/Professor Mimi Tang Allergy and Immune Disorders Infection, Immunity and Environment T 93455911 E mimi.tang@rch.org.au |
Dr Paul Licciardi Allergy and Immune Disorders Infection, Immunity and Environment T 93455554 E paul.licciardi@mcri.edu.au |
Allergic diseases are the commonest chronic illnesses affecting Australian children with over a third suffering from one or more of these disorders. For example, the prevalence of atopic dermatitis (AD, eczema) and asthma among Australian schoolchildren are 20% and 25% respectively, and these are increasing each year. Currently there is no cure for the allergic conditions and management can only control symptoms. Prevention strategies provide a logical approach to reducing the burden of disease in our community. Probiotics offer a promising approach to prevention of allergic disease. Evidence to date suggests that treatment with probiotics can modulate infant microbiota and immune function by resetting the allergic TH2-dominant phenotype towards the healthy TH1 state. A randomised controlled trial has been undertaken (CI: A/Prof Mimi Tang) to evaluate the efficacy of prenatal Lactobacillus GG (probiotic) supplementation to mothers during the last four weeks of pregnancy for the prevention of eczema. This project aims to examine the effect of probiotic supplementation on TH1/TH2 cytokine profiles as well as on the regulatory T cell compartment. Techniques used in this project will include multiplex assay and ELISA.
| A/Professor Mimi Tang Allergy and Immune Disorders Infection, Immunity and Environment T 93455911 E mimi.tang@rch.org.au |
Dr Paul Licciardi Allergy and Immune Disorders Infection, Immunity and Environment T 93455554 E paul.licciardi@mcri.edu.au |
Allergic diseases are the commonest chronic illnesses affecting Australian children with over a third suffering from one or more of these disorders. For example, the prevalence of atopic dermatitis (AD, eczema) and asthma among Australian schoolchildren are 20% and 25% respectively, and these are increasing each year. Currently there is no cure for the allergic conditions and management can only control symptoms. Prevention strategies provide a logical approach to reducing the burden of disease in our community. Prebiotics offer a promising approach to prevention of allergic disease. Evidence to date suggests that treatment with prebiotics can modulate infant microbiota and immune function by resetting the allergic TH2-dominant phenotype towards the healthy TH1 state. A randomised controlled trial has been undertaken to evaluate the efficacy of prebiotic supplemented hydrolysed formula administered to infants for the prevention of eczema. This project aims to examine the effect of prebiotic supplementation on TH1/TH2 cytokine profiles as well as on the regulatory T cell compartment. Techniques used in this project will include multiplex assay and ELISA.
22. Why does this mouse have funny bones?
| Dr Stephanie Gauci Arthritis and Rheumatology Musculoskeletal Disorders T 8341 6431 E steph.gauci@mcri.edu.au |
A/Professor Amanda Fosang Arthritis and Rheumatology Musculoskeletal Disorders T 8341 6466 E amanda.fosang@mcri.edu.au |
The growth of long bones in the skeleton occurs upon a cartilage template in a region called the “growth plate”. Cellular organisation and differentiation within the growth plate is highly structured with quiescent (dormant) cells at the top, proliferative cells in the middle and enlarged, terminally differentiated cells at the bottom. Cell death at the bottom of the growth plate occurs when the cartilage starts to mineralise and is a normal feature of endochondral ossification. In humans, the growth plate disappears at skeletal maturity (about 20 years) and is replaced by bone. This is called growth plate closure. In contrast, the growth plates of rodents persist throughout their lifetime. We have generated a mutant mouse whose cartilage collagen resists enzymatic degradation (Bailey). Bailey mice have elongated growth plates at birth, which persist until 5 weeks of age. However, by 6 weeks, Bailey growth plates are rapidly remodeled and become partially closed. This project will investigate the mechanism(s) of growth plate closure in Bailey using histological and ex vivo culture techniques.
23. Investigating the effects of increased IL-11 expression in the stomach
| Dr Louise Judd Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 99366501 E lmj@unimelb.edu.au |
Dr Trevelyan Menheniott Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 99366502 E treve.menheniott@mcri.edu.au |
| Professor Andrew Giraud T 83416446 E |
Recently our lab has discovered that the cytokine IL-11 has a significant adverse effect on the stomach. In particular we have found that administration of IL-11 to mice for a week results in a condition known as chronic atrophic gastritis. Chronic atrophic gastritis in humans occurs in susceptible individuals following chronic infection with Helicobacter pylori and is a precursor condition to gastric cancer. Helicobacter pylori infection is typically acquired in childhood and it is cumulative exposure that is most pathogenic. To fully understand the effect of IL-11 on the gastric mucosa we have now engineered transgenic mice that over-express IL-11 specifically in the stomach. This project will involve the analysis of these transgenic mice by; handling and dissection of the transgenic mice, analysis of the pathology in the stomach and the rest of the gastrointestinal tract using routine histology and immunohistochemistry and examination of the impact of the IL-11 transgene expression on mRNA and protein expression by Real time PCR and western blotting analysis.
24. Tumour suppressor genes in gastric cancer
| Dr Louise Judd Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 99366501 E lmj@unimelb.edu.au |
Dr Trevelyan Menheniott Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 99366502 E treve.menheniott@mcri.edu.au |
| Professor Andrew Giraud T 83416446 E |
Trefoil 1 (TFF1) is a tumour suppressor gene that has been shown to have decreased expression in human gastric and breast cancer through gene methylation. In our lab we have a mouse model of gastric tumours the gp130757FF model that has an engineered mutation in the cytokine rector gp130 which results in the 100% of mice carrying the mutation developing tumours by 8 weeks of age. While these gastric tumours mimic many of the characteristics of human disease they never become metastatic a hallmark of human gastric tumours. The gp130757FF mouse has low to moderate expression of TFF1 in the stomach. Recently we have obtained mice with a knockout mutation in TFF1, these mice also spontaneously develop gastric tumours but not until 1 year of age. In this project we will generate mice that have both the gp130757FF mutation and theTFF1 to determine how the absence of the tumour suppressor gene TFF1 impacts on gp130757FF gastric tumours. Analysis of these knockout mice will be by; handling and dissection of the mice, analysis of the pathology in the stomach and the rest of the gastrointestinal tract using routine histology and immunohistochemistry and examination of the impact of the engineered mutations on mRNA and protein expression by Real time PCR and western blotting analysis.
25. Identification of Rotavirus vaccine strains in children with gastroenteritis.
| Dr Carl Kirkwood Enteric Viruses Infection, Immunity and Environment T 8341 6439 E carl.kirkwood@mcri.edu.au |
Dr Jim Buttery Enteric Viruses Infection, Immunity and Environment T 9345 4772 E jim.buttery@mcri.edu.au |
Rotavirus is the major cause of acute gastroenteritis in children in developed and developing countries, causing over 500,000 deaths annually. In Australia over 10,000 children are admitted to hospital each year. To reduce the disease burden two rotavirus vaccines (Rotarix and RotaTeq) have been developed and licensed. These vaccines were introduced into the routine immunisation schedule in Australia for all newborn infants on 1st July 2007. Rotarix and RotaTeq strains are shed in infants administered vaccine (50% and 10% after first dose), and a small percentage of these infants develop diarrhoea. The vaccine strains may also enter the community and circulate with wildtype strains causing disease. This research project is designed to characterise rotavirus vaccine strains in vaccinated infants and identify vaccine strains circulating in the community. The project will involve a range of molecular biology techniques such as RT-PCR, Northern hyridisation and sequence analysis, aimed to determine whether the isolates characterised are vaccine-like or wildtype.
26. Understanding the role of infectious agents in children with early onset Crohn’s disease
| Dr Carl Kirkwood Enteric Viruses Infection, Immunity and Environment T 8341 6439 E carl.kirkwood@mcri.edu.au |
Dr Josef Wagner Enteric Viruses Infection, Immunity and Environment T 8341 6450 E josef.wagner@mcri.edu.au |
Crohn’s disease is a major cause of morbidity throughout the world. It is an incurable condition associated with chronic inflammation of the gastrointestinal tract of genetically susceptible individuals. Crohn’s disease usually begins in early adulthood and is treated with potent immunosuppressive medications, but often requires surgery. We have conducted preliminary studies designed to identify an infectious agent in gut biopsy tissue obtained from children with suspected Crohn’s disease. Using microarray and subtractive hybridisation techniques we have identified a infectious agents (bacteria & viruses) that we propose could initiate the gut damage and ongoing immune activation. This project will continue to utilise molecular techniques to genetically characterise the agents in children with Crohn’s disease and develop specific PCR detection assays to determine the prevalence in clinical specimens collected from children at disease onset.
27. Childhood Arthritis – the role of epigenetics in determining risk of disease
| Dr Justine Ellis Environmental and Genetic Epidemiology Research Infection, Immunity and Environment T 83416311 E justine.ellis@mcri.edu.au |
Dr Jeff Craig Developmental Epigenetics Early Development and Disease T 83416346 E jeff.craig@mcri.edu.au |
The factors that lead to development of arthritis in childhood (juvenile idiopathic arthritis - JIA), are currently largely unknown. However, this disease can have devastating effects, not only due to the pain and disability that is experienced, but also by affecting normal growth and development. It is believed that JIA is caused by a combination of genetic and environmental factors, and these are the focus of research within our CLARITY JIA Biobank. A potential risk factor yet to be considered in relation to JIA is that of epigenetic variation. In this project we will consider the question of whether differences in methylation occur across the genome when DNA from JIA patients and healthy control children are compared. We shall use data obtained from genome-wide methylation arrays to look for potential points of difference, then follow up this hypothesis-generating data by conducting in depth analyses if specific genes that appear differentially methylated. The project will potentially involve exposure to patient recruitment, biospecimen processing, cell isolation and sorting, DNA extraction, PCR, methylation analysis using the Sequenom MassARRAY system, bioinformatic and statistical approaches to data analysis.
| Dr Matthew Sabin Hormone Research Early Development and Disease T 93456986 E matthew.sabin@mcri.edu.au |
Dr Richard Saffery Developmental Epigenetics Early Development and Disease T 83416341 E richard.saffery@mcri.edu.au |
Childhood obesity is associated with the development of metabolic complications, such as Type 2 diabetes. While environmental factors and specific genetic traits are known to influence weight gain and metabolic health, it is still unclear how environment and genetics come together to influence risk of disease. A likely candidate mediating these combined effects is epigenetics, involving specific modifications to genes within the body that regulate their expression independently of primary DNA sequence. The country’s largest specialist paediatric weight management service is run from the Royal Children’s Hospital in Melbourne, and children and adolescents attending this service are now routinely entered into COBRA - the Childhood Overweight BioRepository of Australia. As part of this project, comprehensive environmental and clinical data are collected, along with peripheral blood for metabolite and molecular analyses. The current research project will specifically explore the epigenetic regulation of genes postulated to control metabolism and body weight homeostasis. It is anticipated that this will provide valuable insight into the unique ways in which diet programs weight regulation over time, independent of short term caloric intake.
29. Analysis of mouse models of human birth defects
| Dr Peter Farlie Craniofacial Research Musculoskeletal Disorders T 83416409 E peter.farlie@mcri.edu.au |
Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
Birth defects involving the face affect approximately 1% of all babies but the genes involved in most of these conditions are unknown. ENU mutagenesis is a forward genetics approach that allows phenotypes of interest to be identified without any prior knowledge of specific genes involved in the development of the organ system of interest. Once useful phenotypes are established, rapid gene mapping approaches have been developed to identify the mutated gene. We have a number of mouse strains harbouring birth defects involving both the face and limbs that appear similar to human syndromes. Identification of the mutated genes in these strains will help us understand how the normal developmental program is altered in human birth defects and will facilitate identification of disease genes in humans. This project will provide an opportunity for students to learn developmental biology approaches to understanding human birth defects and gain skills in a wide range of molecular biology techniques. Upon completion of this project, students will be in a strong position to initiate their own research into the genetic and developmental basis of human birth defects.
30. Large-scale screen of genes controlling skeletal development
| Dr Peter Farlie Craniofacial Research Musculoskeletal Disorders T 83416409 E peter.farlie@mcri.edu.au |
Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
Congenital defects of the skeleton are common and have a major impact on health and well being of affected children. Microarray RNA expression analysis of skeletal development is a powerful genome scale screening technology that is beginning to reveal essential pathways in skeletogenesis. However, a major limitation in this process is the functional analysis of identified candidate genes. To address this limitation in the analysis of our microarray data, we have developed a high-throughput screen to analyse the function of candidate genes in early skeletal development using avian retroviral delivery of expression and knockdown constructs. This screen will allow the student to rapidly analyse gene function in a whole animal model and will facilitate large-scale functional analysis of the genes controlling skeletal development and causing human skeletal defects and disease. Students will use cutting edge approaches to dissecting the genetic networks controlling complex developmental events during formation and growth of the craniofacial and limb skeletons. Experience gained in this project will allow students to initiate investigations into the mechanisms controlling development of any organ system.
31. Analysis of mouse models of human birth defects
| Dr Peter Farlie Craniofacial Research Musculoskeletal Disorders T 83416409 E peter.farlie@mcri.edu.au |
Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
Birth defects involving the face affect approximately 1% of all babies but the genes involved in most of these conditions are unknown. ENU mutagenesis is a forward genetics approach that allows phenotypes of interest to be identified without any prior knowledge of specific genes involved in the development of the organ system of interest. Once useful phenotypes are established, rapid gene mapping approaches have been developed to identify the mutated gene. We have a number of mouse strains harbouring birth defects involving both the face and limbs that appear similar to human syndromes. Identification of the mutated genes in these strains will help us understand how the normal developmental program is altered in human birth defects and will facilitate identification of disease genes in humans. This project will provide an opportunity for students to learn developmental biology approaches to understanding human birth defects and gain skills in a wide range of molecular biology techniques. Upon completion of this project, students will be in a strong position to initiate their own research into the genetic and developmental basis of human birth defects.
32. Large-scale screen of genes controlling skeletal development
| Dr Peter Farlie Craniofacial Research Musculoskeletal Disorders T 83416409 E peter.farlie@mcri.edu.au |
Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
Congenital defects of the skeleton are common and have a major impact on health and well being of affected children. Microarray RNA expression analysis of skeletal development is a powerful genome scale screening technology that is beginning to reveal essential pathways in skeletogenesis. However, a major limitation in this process is the functional analysis of identified candidate genes. To address this limitation in the analysis of our microarray data, we have developed a high-throughput screen to analyse the function of candidate genes in early skeletal development using avian retroviral delivery of expression and knockdown constructs. This screen will allow the student to rapidly analyse gene function in a whole animal model and will facilitate large-scale functional analysis of the genes controlling skeletal development and causing human skeletal defects and disease. Students will use cutting edge approaches to dissecting the genetic networks controlling complex developmental events during formation and growth of the craniofacial and limb skeletons. Experience gained in this project will allow students to initiate investigations into the mechanisms controlling development of any organ system.
33. Lung function genes and airway remodelling in asthma.
| Dr Simon Royce Allergy and Immune Disorders Infection, Immunity and Environment T 93456882 E simon.royce@mcri.edu.au |
A/Professor Mimi Tang Allergy and Immune Disorders Infection, Immunity and Environment T 93455911 E mimi.tang@rch.org.au |
Asthma is the commonest chronic disease of Australian children and although generally well managed, asthma still accounts for 1 in 250 deaths. A subset of asthma sufferers is unresponsive to current therapies and develops structural changes in the airways that worsen lung function. We are interested in developing new therapies for asthma that target airway remodelling. Two endogenous peptides (relaxin and tff2) have been extensively studied in mouse knockout and allergic airways disease models. We now wish to investigate their role in human asthma using archival tissue samples, and lung cell lines, including primary cells from patients with asthma. Antibody based and molecular techniques will be used in the investigation and results will be correlated with clinicopathological data already collected as part of a large asthma study.
34. Epithelial remodelling and repair in airway disease.
| Dr Simon Royce Allergy and Immune Disorders Infection, Immunity and Environment T 93456882 E simon.royce@mcri.edu.au |
A/Professor Mimi Tang Allergy and Immune Disorders Infection, Immunity and Environment T 93455911 E mimi.tang@rch.org.au |
Airway remodelling refers to the structural changes that occur in the airways of asthmatics. During the pathogenesis of this disease airway remodelling results in thickening of the airway wall, and the consequent reduction in luminal diameter results in worsened lung function and susceptibility to death. We will be testing the hypothesis that there is an inherent defect in the epithelium in asthma and investigating how this defect in epithelial repair drives other remodelling changes. Knockout and epithelial damage mouse models of asthma, asthma cell and tissue samples will be probed for remodelling parameters using techniques including morphometry, immunohistochemistry, ELISA, molecular biology and cell culture. This investigation has direct links with other studies underway in the laboratory to develop new therapies for airway remodelling in asthma.
35. The molecular signalling pathways that cause osteoarthritis
| Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 8341 6422 E john.bateman@mcri.edu.au |
Dr Richard Wilson Skeletal Biology and Disease Musculoskeletal Disorders T 8341 6418 E richardwilson.m@mcri.edu.au |
Degeneration of articular cartilage is the central pathological feature of osteoarthritis (OA) and it is this progressive erosion of cartilage that leads to joint failure and necessitates joint replacement surgery. We have a major research program determining the molecular events in the initiation and progression of cartilage breakdown. Using microarray analysis to look at gene expression changes and proteomic approaches we have determined new osteoarthritis candidate genes. Several research projects are available in this program exploring the detailed biology of these genes and the signalling pathways that result in the onset and progression of OA. These studies will involve the study of the biological function of these genes in cultured cartilage and bone cells, and how both over-expression and expression knockdown by RNAi affects cellular signalling and cell phenotype in vitro. This will compared with the changes found in osteoarthritic tissues. The projects will use of a wide range of molecular biology, biochemical, cell biology and proteomic techniques.
| Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
Dr Richard Wilson Skeletal Biology and Disease Musculoskeletal Disorders T 93456601 E richardwilson.m@mcri.edu.au |
| Mr Trevor Cameron T 93456602 E |
Inherited musculoskeletal disorders are a significant disease burden and although many mutations have been defined, our knowledge on the molecular mechanisms that cause them, and ultimately how these mechanisms could be manipulated, is only just beginning to be explored. Many of the gene mutations result in the production of mutant protein that is compromised in its ability to form the correct 3-D folded functional structures. Recent research has shown that these unfolded proteins can cause cellular stress and activate intracellular signalling pathways that have profound effects on cell gene expression that may contribute to cellular pathology (see Bateman et al.,Nature Reviews Genetics 2009 Mar;10(3):173-83). The proposed studies will explore the molecular signalling pathways using in vitro and transgenic mouse models and a range of immunohistochemical, biochemical, molecular methods and proteomic analysis (2D-electrophoresis and mass spectrometry) skills. In addition, our studies will explore the use of new therapeutic agents to overcome protein misfolding and cell stress, as a proof-of-principle that some of these diseases can be effectively treated.
37. Identifying TRPV4 interacting partners critical in human disease phenotypes
| Dr Shireen Lamande Muscular Dystrophy Musculoskeletal Disorders T 83416465 E shireen.lamande@mcri.edu.au |
Dr Laura Zamurs Muscular Dystrophy Musculoskeletal Disorders T 83416418 E laura.zamurs@mcri.edu.au |
Mutations in the calcium channel TRPV4 cause defective skeletal development and peripheral nerve defects. Consistent with the disease phenotypes, TRPV4 is expressed in cartilage and bone cells, but it is also found in many other tissues including nerves and kidney and we don’t yet understand why mutations predominantly affect either the skeleton or nerves. We hypothesise that the neuropathy and skeletal dysplasia mutations affect different signalling pathways and different protein-protein interactions but very little is known about these pathways and so TRPV4 interacting proteins will be identified in a yeast two-hybrid screen. The project will also aim to characterise the interactions with wild type and mutant TRPV4 proteins, and will examine expression of interacting proteins in affected tissues and cells.
38. TRPV4 and skeletal development
| Dr Shireen Lamande Muscular Dystrophy Musculoskeletal Disorders T 83416465 E shireen.lamande@mcri.edu.au |
Professor John Bateman Skeletal Biology and Disease Musculoskeletal Disorders T 83416422 E john.bateman@mcri.edu.au |
| Ms Lynn Rowley T 93456601 E |
Mutations in the calcium channel TRPV4 cause defective skeletal development but very little is known about the temporal and spatial expression of TRPV4 during cartilage and bone development. This project will characterise TRPV4 mRNA and protein expression during skeletal development in the mouse using in situ hybridization and immunohistochemistry. We know that the Trpv4 knockout mouse has some abnormal skeletal features but to date only the knee joints have been studied and so another aspect of this project will involve detailed analyses of the skeleton in this mouse during development using histology and a range of imaging techniques.
39. Collagen VI and WARP interactions during neurological development
| Dr Shireen Lamande Muscular Dystrophy Musculoskeletal Disorders T 83416465 E shireen.lamande@mcri.edu.au |
Ms Lynn Rowley Skeletal Biology and Disease Musculoskeletal Disorders T 93456601 E lynn.rowley@mcri.edu.au |
| Professor John Bateman T 83416422 E |
WARP is a recently identified extracellular matrix molecule with expression restricted to permanent cartilages and a distinct set of basement membranes in peripheral nerves, muscle, and the central nervous system vasculature. WARP knockout mice are healthy, viable, and fertile with no overt abnormalities; however, they have a significantly delayed response to acute painful stimulus and impaired fine motor coordination, suggesting compromised peripheral nerve function. WARP interacts with the extracellular matrix protein collagen VI and the collagen VI microfibrillar network is severely reduced and mislocalised in peripheral nerves of WARP knockout mice. Collagen VI is broadly distributed and mutations in humans cause muscular dystrophy. Other tissues such as cartilage, bone and tendons are also affected although these phenotypes have been poorly defined. Our data suggests that collagen VI may also be important in peripheral nerves and this project will define the role of collagen VI in the nervous system using collagen VI knockout mice, WARP knockout mice and collagen VI/WARP double knockout mice. This project will predominantly involve histology, immunostaining and electron microscopy.
40. Stomach inflammation and epigenetic silencing of the TFF2 tumour suppressor gene
| Dr Trevelyan Menheniott Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 03 9936 6502 E treve.menheniott@mcri.edu.au |
Dr Louise Judd Gastrointestinal Research in Inflammation and Pathology (GRIP) Infection, Immunity and Environment T 03 9936 6501 E lmj@unimelb.edu.au |
| Professor Andrew Giraud T 03 8341 6446 E |
Over one million deaths from stomach cancer will occur annually in the next decade. This often fatal disease can be caused by infection from childhood by the bacterium Helicobacter pylori (H. pylori). During the early stages of the disease, inflammation of the stomach epithelium caused by H. pylori infection initiates precancerous changes which can progress to tumour formation. Using a range of mouse infection models, this project aims to investigate the role of H. pylori-dependent inflammation in driving epigenetic silencing via DNA methylation of the stomach tumour suppressor gene, TFF2. This new information may enable us to design new approaches for early detection and treatment of stomach cancer. Techniques: Transgenic/knockout mouse analysis, DNA methylation analysis, quantitative RT-PCR, western blotting. Animal ethics approval has been obtained (A633).
41. Epigenetic factors in Triplet-Repeat Associated Diseases
| Dr Richard Saffery Developmental Epigenetics Early Development and Disease T 83416341 E richard.saffery@mcri.edu.au |
Dr Marguerite Evans-Galea Genetic Health Research (Bruce Lefroy Centre) Laboratory and Community Genetics T 83416286 E marguerite.galea@mcri.edu.au |
| A/Professor Martin Delatycki T 83416284 E |
The importance of epigenetic changes in neurodegenerative disease, and their potential influence on clinical parameters, is just being realised. Several conditions, such as Friedreich ataxia, Fragile-X syndrome, Kennedy disease and Huntington disease, are caused by trinucleotide repeat expansions in specific genes. Recent work within the MCRI has established a link between DNA methylation in the region of repeat expansion with clinical parameters of Fragile X Syndrome and Friedreich ataxia. In this study, we will employ a range of molecular and cellular techniques (PCR, quantitative gene expression, DNA methylation analysis by bisulfite sequencing and MALDI-TOF mass spectrometry) to examine the correlation of various clinical parameters to expansion size and DNA methylation patterns in Kennedy and Huntington diseases. These studies will provide valuable insights into the role of epigenetic disruption in modulating the clinical phenotype in repeat-associated neurodegenerative diseases and will identify potential prognostic biomarkers for use in the clinic.
42. Modifying and Preventing Heart Disease in Mice with Mitochondrial Cardiomyopathy
| A/Professor David Thorburn Mitochondrial Research Laboratory and Community Genetics T 83416235 E david.thorburn@mcri.edu.au |
Dr Bi-Xia Ke Mitochondrial Research Laboratory and Community Genetics T 83416287 E bi-xia.ke@mcri.edu.au |
| Dr Salvatore Pepe T 93454114 E |
Genetic disorders of mitochondrial energy generation cause a range of severe diseases in at least 1/5000 individuals. Milder genetic or acquired mitochondrial dysfunction contributes to common diseases including diabetes and heart disease. We generated one of the first mouse models of mitochondrial complex I deficiency by knockdown of the Ndufs6 gene. The mice have gross Complex I deficiency in the heart, which becomes enlarged, more markedly in males than females. We have now generated congenic Ndufs6 mice on C57BL/6 and BALB/c backgrounds. Preliminary results suggest that the cardiac dysfunction is more severe in C57BL/6 mice than on a mixed background and we predict that one factor contributing to this is the absence of Nicotinamide Nucleotide Transhydrogenase (NNT) activity in C57BL/6 mice. This project will complete characterization of the cardiac phenotype of mice on different genetic backgrounds using a range of echocardiographical, histological, metabolic, physiological, molecular and immunochemical approaches. We will investigate the role of NNT in modifying the phenotype. In addition, therapeutic trials will be carried out on mice using bezafibrate, a drug that can stimulate mitochondrial biogenesis to upregulate transcription of respiratory chain genes. Our expectation is that this project will lead to a better understanding of pathogenic mechanisms of complex I deficiency and improvement of treatment strategies.