2022 SBSP BACHELOR OF BIOMEDICAL SCIENCES HONOURS PROJECT Page | 1 PROJECT DETAILS A) Project Title: Investigating molecular mechanisms of a diet and resistance exercise program to prevent sarcopenia in the elderly. B) Supervisory Details: Primary Supervisor Name: Professor Lisa Wood Location: HMRI, Level 2 – West / Room 606, Medical Sciences Building Email: [email protected]Phone: 49217485 Co-Supervisor Name: Dr Evan Williams Location: HMRI, Level 2 – West Email: [email protected]Phone: (02) 404 20910 Co-Supervisor Name: Dr Hayley Scott Location: HMRI, Level 2 – West Email: [email protected]Phone: (02) 4042 0113 C) Background and Summary of Proposed Research, including a clear hypothesis, aims and experimental approach: The loss of muscle mass, strength and function as we age (sarcopenia) is associated with increased risk of falls, hospitalisation and reduced independence. Sarcopenia is a common comorbidity of arthritis and osteoporosis, all of which can accelerate age-related functional decline. We recently completed a randomised controlled trial investigating whether improving nutrition and home-based resistance training in older untrained adults during a 16 week period can improve outcomes such as muscle mass, bone density, grip strength and gait speed. This project will investigate the molecular pathways modulated by the intervention using Affymetrix Microarray (Clariom S) to measure peripheral blood gene expression. Bioinformatics tools will be used for network and pathway analysis. The student will develop unique experience in molecular biology laboratory techniques and bioinformatics tools used in gene profiling. HYPOTHESIS: Improvements in muscle mass, bone mineral content, muscle strength and function following a 16-week combined exercise and nutrition intervention in adults at risk of sarcopenia, are associated with differential gene expression of immune and metabolic pathways. AIMS: To use transcriptome and network analysis to identify changes in immune and metabolic pathways following a 16- week combined exercise and nutrition intervention in adults at risk of sarcopenia, and to examine the relationship between these changes and improvements in muscle mass, bone mineral content, muscle strength and function. METHOD: This study will utilise previously collected microarray data from an RCT in adults at risk of sarcopenia. They will perform transcriptome analysis using microarray data, gene ontology and pathways analysis using computer programs. The student will then confirm these findings in the lab by performing qPCR for gene expression and Enzyme-linked Immunoassay (ELISA) to confirm protein expression. Laboratory and statistical techniques: Microarray data will be analysed using R studio. Relationships of gene expression profiles will be examined using hierarchical clustering. Potential molecular mechanisms will be investigated by both gene ontology and pathways analysis using STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), GATHER (Gene Annotation Tool to Help Explain Relationships) and KEGG (Kyoto Encyclopedia of Genes and Genomes). These findings will then be pursued in the lab by measuring gene expression by qPCR and protein expression by ELISA. Associations between clinical improvements and changes in molecular pathways will be examined using multiple regression. D) Laboratory Location Wood Lab: HMRI, Level 2 – West Wing
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2022 SBSP BACHELOR OF BIOMEDICAL SCIENCES HONOURS PROJECT
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PROJECT DETAILS
A) Project Title: Investigating molecular mechanisms of a diet and resistance exercise program to prevent sarcopenia
in the elderly.
B) Supervisory Details:
Primary Supervisor Name: Professor Lisa Wood
Location: HMRI, Level 2 – West / Room 606, Medical Sciences Building
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and experimental approach:
The loss of muscle mass, strength and function as we age (sarcopenia) is associated with increased risk of falls, hospitalisation and reduced independence. Sarcopenia is a common comorbidity of arthritis and osteoporosis, all of which can accelerate age-related functional decline. We recently completed a randomised controlled trial investigating whether improving nutrition and home-based resistance training in older untrained adults during a 16 week period can improve outcomes such as muscle mass, bone density, grip strength and gait speed. This project will investigate the molecular pathways modulated by the intervention using Affymetrix Microarray (Clariom S) to measure peripheral blood gene expression. Bioinformatics tools will be used for network and pathway analysis. The student will develop unique experience in molecular biology laboratory techniques and bioinformatics tools used in gene profiling.
HYPOTHESIS: Improvements in muscle mass, bone mineral content, muscle strength and function following a 16-week
combined exercise and nutrition intervention in adults at risk of sarcopenia, are associated with differential gene expression
of immune and metabolic pathways.
AIMS: To use transcriptome and network analysis to identify changes in immune and metabolic pathways following a 16-
week combined exercise and nutrition intervention in adults at risk of sarcopenia, and to examine the relationship between
these changes and improvements in muscle mass, bone mineral content, muscle strength and function.
METHOD: This study will utilise previously collected microarray data from an RCT in adults at risk of sarcopenia. They will perform transcriptome analysis using microarray data, gene ontology and pathways analysis using computer programs. The student will then confirm these findings in the lab by performing qPCR for gene expression and Enzyme-linked Immunoassay (ELISA) to confirm protein expression.
Laboratory and statistical techniques: Microarray data will be analysed using R studio. Relationships of gene expression profiles will be examined using hierarchical clustering. Potential molecular mechanisms will be investigated by both gene ontology and pathways analysis using STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), GATHER (Gene Annotation Tool to Help Explain Relationships) and KEGG (Kyoto Encyclopedia of Genes and Genomes). These findings will then be pursued in the lab by measuring gene expression by qPCR and protein expression by ELISA. Associations between clinical improvements and changes in molecular pathways will be examined using multiple regression.
D) Laboratory Location
Wood Lab: HMRI, Level 2 – West Wing
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PROJECT DETAILS Project Title: Promoting oligodendrocyte maturation in the preterm neonatal brain
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and experimental
approach:
We have successfully completed a 6-month randomised controlled trial (RCT) to improve diet quality in children with asthma. Following intervention, lung function improved and the number of children who had multiple asthma attacks in a 6-month period declined. It is hypothesised that the positive results seen in this trial may have been mediated by changes in the immune system.
This project is a molecular investigation into immune pathways modulated by our intervention, including gene expression analysis of peripheral blood mononuclear cell samples using Nanostring and associated bioinformatics tools to complete network and pathway analysis. The student will develop unique experience in molecular biology laboratory techniques and bioinformatics tools used in gene profiling.
HYPOTHESIS: Improvements in lung function and reduced asthma-related illness following a dietary intervention are
associated with differential expression of immune system pathways.
AIMS: To use transcriptome and network analysis to identify changes in immune pathways following a dietary intervention
in children with asthma, and to examine the relationship between these changes and improvements in lung function and
exacerbation frequency.
METHOD: This study will utilise stored peripheral blood mononuclear cell (PBMC) samples previously collected during an RCT in children with asthma. The student will perform the RNA extractions, transcriptome analysis using Nanostring assays, and gene ontology and pathways analysis using computer programs.
Laboratory and statistical techniques: RNA will be extracted from PBMCs using the RNeasy Mini Kit (Qiagen, Hilden, Germany), quantitated using the Quant-iT RiboGreen RNA Assay Kit (Molecular Probes Inc, Invitrogen, Eugene, OR, USA). PBMC transcriptome analysis will be performed in baseline and 6-month samples using the Nanostring nCounter Analysis System Human Immunology v2 Panel (Nanostring Technologies, Seattle, WA, USA), where expression of 579 immunology-related genes and 15 internal reference controls will be measured. Nanostring data will be analysed using nSolver Analysis Software v2.5 (Nanostring Technologies). Gene profiles will be analysed for differential expression by paired t-test for significance. Relationships of gene expression profiles will be examined using hierarchical clustering. Potential molecular mechanisms will be investigated by both gene ontology and pathways analysis using STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), GATHER (Gene Annotation Tool to Help Explain Relationships) and KEGG (Kyoto Encyclopedia of Genes and Genomes). Associations between clinical improvements and changes in inflammatory pathways will be examined using multiple regression.
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Background: Asthma affects 300 million individuals world-wide, and is characterised by recruitment of immune cells to the lungs and airways hyperresponsiveness. The 2019-2020 bushfire season resulted in a thick blanket of smoke descending on millions of Australians for weeks and months at a time without respite, causing asthmatic exacerbations and resulting in increased hospital admissions. We have developed a novel in vivo model of bushfire smoke exposure that recapitulates the features of particulate matter-induced asthmatic exacerbations such as airways hyperresponsiveness, and aberrant proinflammatory gene expression in the lungs. Importantly, there is a noted dynamic between neurobiological and immunological mechanisms in the lungs and this interplay, in the context of asthma, will be the focus of this project. Hypothesis: Bushfire smoke exposure induces neurobiological changes within the small airways of the lungs, with altered afferent and efferent communication contributing to airways hyperresponsiveness and mediation of immune cell interactions. Aim: To investigate the mechanisms by which bushfire smoke particulate exposure alters afferent and efferent nerve fibres in the small airways, and how these alterations contribute to the inflammatory profile observed in asthmatic lungs. Methods: The successful applicant will employ our novel bushfire smoke exposure models in their research. Highly specialised research techniques will be used such as precision cut lung slicing (PCLS) to investigate mechanisms of particulate matter-induced airways hyperresponsiveness. Sectioned and stained tissues will be used to determine histological features of disease such as collagen deposition and mucus secretion. Immunohistochemistry (IHC) and immunofluorescence (IF) will be employed to detect and localise factors of interest within histological sections, and highly specialised equipment and techniques will be used to measure lung function outcomes.
D) Laboratory Location: Hunter Medical Research Institute (HMRI).
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
The 2019 bushfires had a devastating effect and for months exposed people on the Eastern seaboard of
Australia to high levels of air pollution. We are part of a successful MRFF grant application to assess the impact
of bushfire smoke exposure on the lungs and immune system. We will elucidate the short and prolonged
physiological effects of bushfire smoke from different areas of Australia using our unique primary human cells
models of disease. We will assess the impact on cells from people with pre-existing chronic respiratory
diseases (asthma, COPD) and at different ages (pregnancy, infancy, elderly), and define safe exposure levels.
We will use a world first device to aerosolise the smoke and expose these cells to elucidate cell and tissue and
responses.
Our hypothesis is that bushfire smoke (BFS) promotes inflammation and damages airway epithelia, which is
exacerbated in patients with chronic respiratory diseases and differs by age.
To address this, the specific project aims will be:
1) Identify differences in the effects of BFS from different states on inflammatory responses and histopathological changes in primary bronchial epithelial cells from heathy human adults during acute (1-3 days) and chronic (14 days) exposures.
2) Characterise the effects of BFS on from human on inflammatory responses and histopathological changes in primary bronchial epithelial cells from COPD and asthma patients during acute (1-3 days) and chronic (14 days) exposures.
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Due to our ageing population, osteoporotic fractures are on the rise and pose a significant burden to the
healthcare system and society, with a hip fracture occurring every 3 minutes in Australia and > $2
billion spent annually in Australia on fracture treatment. Time to union is longer with increasing age
and in the presence of osteoporosis. Delayed and non-union are associated with significant morbidity,
multiple unplanned hospitalisations, loss of mobility and independence, all of which increases mortality
in elderly. The underlying mechanisms for poor fracture healing in the elderly are poorly understood.
Following bone fracture, mesenchymal stem cells (MSCs) are the source of new bone, where they enter
the fracture site, proliferate and differentiate into cartilage producing cells (chondrocytes) and bone-
producing cells (osteoblasts). Both aged and osteoporotic fracture healing has been related to a reduced
number of MSCs, and a reduced and delayed ability to differentiate into chondrocytes and osteoblasts.
Aged MSCs and osteoblasts have been shown to have impaired mitochondrial function, however this
data has used animal models and to date there are no investigations of mitochondrial dysfunction in
elderly human MSCs. We hypothesise that MSCs isolated from elderly hip fracture patients will
have reduced mitochondrial function.
This project will be based at the John Hunter Hospital in the Surgical Sciences Lab. In this setting, we have access to bone marrow samples removed routinely during hip fracture surgery. The honours
student will test the above hypothesis by culturing MSCs from these samples in the laboratory,
measuring mitochondrial mass, membrane polarisation, OXPHOS and ATP production. In parallel, a
control group of bone marrow samples from young patients requiring surgical management of fractures
will be collected.
D) Laboratory Location
Surgical Sciences Laboratory, John Hunter Hospital
B) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Summary Placental support of fetal growth and development in-utero are essential for lifelong health. Central to sufficient placental function is mitochondria, facilitating the metabolic changes, substrate utilisation and oxygen supply to the growing fetus. Detrimental changes in placental function involving mitochondria insufficiency is associated with many pregnancy complications including stillbirth and ongoing metabolic syndromes that lead to disability later in life. Despite this critical role, little is known about the metabolic changes across gestation, and how the mitochondria within the placenta facilitate and adapt to the altering environment to optimise healthy outcome. Aims This project will examine placental metabolism across gestation, and establish how the mitochondria differ between the first trimester and third trimester. Subsequently, this study will investigate the mechanisms which underpin the observed metabolic changes that lead to poor outcomes. Hypothesis We suspect that metabolic processes present in the first trimester are significantly different than those observed in the third trimester. Specifically, that first trimester metabolism will favour aerobic glycolysis over oxidative phosphorylation and alter their metabolic pathways accordingly. Experimental approach This project will be utilising new state of the art measures to assess metabolism and mitochondrial function, including real-time functional respiration (Agilent Seahorse), plate-based enzyme activity assays in addition to assessing gene expression and proteins. This will enable evaluation of the bioenergetic capacity and provide clear assessment of metabolic pathways. This project will provide the opportunity to develop a fundamental knowledge of metabolism and mitochondrial physiology, and develop skills in numerous laboratory techniques.
C) Laboratory Location
HMRI Level 3 East
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PROJECT DETAILS
A) Project Title: Targeting long noncoding RNAs to sensitize cancer to metabolic inhibition.
B) Supervisory Details :
Primary Supervisor Name: Xu Dong Zhang
Location: LS3-49 Life sciences building, University Drive, Callaghan, NSW 2308, Australia
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Long nonprotein-coding RNAs (lncRNAs) were historically thought to be transcriptional noise without biological function. However, it is now clear that they function to regulate diverse pathophysiological processes. Our previous studies have unveiled a number of lncRNAs which play important roles in cancer initiation, development and drug resistance. Targeting cancer cell metabolism has been emerging as a strategy for cancer treatment. Numerous studies have demonstrated that the mitochondrial oxidative phosphorylation (OXPHOS) system is necessary for cancer cell proliferation and survival. Therefore, OXPHOS has become a potential therapeutic target for cancer treatment. Indeed, small molecule OXPHOS inhibitors have already entered clinical evaluation. However, cancer cell resistance and adaptation to OXPHOS inhibition occurs. In this project, we’d like to explore lncRNA roles in the resistance of cancer cell to metabolic inhibition.
We have tested the responses of a panel of colorectal cancer (CRC) cell lines to an OXPHOS inhibitor, IACS-010759 (IACS), which inhibits mitochondrial complex I. Among them, we found that LIM1215 and Caco-2 cell lines were resistant to IACS treatment. Further RNA sequencing analysis identified a list of lncRNAs that were upregulated upon IACS treatment in IACS-resistant cell lines, suggesting the potential roles of these lncRNAs in protecting CRC cells from OXPHOS inhibition.
In this project, we aim to clarify lncRNA-mediated mechanisms responsible for the resistance of CRC cells to IACS treatment. The expression of candidate lncRNAs in cells before and after treatment with IACS will be tested using qPCR. Their functions will be examined using siRNA/shRNA and cDNA transfection/transduction, followed by cell viability, cell death and clonogenicity assays. LncRNA-binding proteins will be identified using RNA pulldown followed by mass spectrometry analysis. The interaction between lncRNA and proteins will be confirmed using RNA pulldown and RNA immunoprecipitation assays. These results will potentially identify lncRNAs that protect CRC cells from OXPHOS inhibition and develop RNA therapeutics by targeting lncRNAs as a strategy to sensitize CRC cells to OXPHOS inhibition.
D) Laboratory Location: LS3-20 Life sciences building, University Drive, Callaghan, NSW 2308.
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PROJECT DETAILS
A) Project Title: A genomic editing approach for indication the activation status of the autonomous
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Colorectal cancer (CRC) is the third most commonly diagnosed cancer and the second most common cause of cancer death in Australia. Elimination of metastases, but not removal of the primary tumours, is the biggest obstacle to reduce the mortality caused by CRC. Chemotherapy is recommended in most metastatic cases as surgery is not possible to clear lesions at multiple sites. Of note, during the metastatic cascade, cancer cells tightly interact with the immune system. The outcome of chemotherapy can be influenced by the host immune system at multiple levels. Moreover, autonomous interferon signalling in cancer cells is potentially a trigger of the anti-cancer immune response and can also inhibit cell proliferation and induce cell death, and thus contribute to the efficacy of treatments. Of note, oxaliplatin is a first-line drug in the treatment of late stage of the disease, which were associated with activation of type I and type II interferon signalling.
An interferon-stimulated response element (ISRE) is a conserved nucleotide sequence responsible for the activity of Type I interferon-induced JAK/STAT signalling pathway. In this honours student project, we will develop an approach based on CRISPR/Cas9 technology to knock-in an enhanced green fluorescence protein (EGFP) gene under the control of multimerized ISRE into the safe harbor region in CRC cell lines (ISRE/EGFP cells) so that the activation status of the autonomous interferon signalling in the CRC cells could be visualised, tracked, and analysed by a flow cytometer or a fluorescence microscope based on the GFP levels.
A PhD project could be logically extended from this ISRE/GFP cell line, which we will further adapt a functional genetic screening platform to identify specific molecules that ignite the autonomous interferon signalling when the gene(s) is/are silenced in the cells based on this fluorescence system. The identified candidates will be further tested for development of novel approaches in the treatment of metastatic CRC.
In this project, we hypothesize that the ISRE/EGFP cells could indicate the activation status of autonomous interferon signalling and could be employed for the following functional screening. Our specific aims are: 1) To establish CRC cell lines containing multimerized ISRE controlled EGFP by CRISPR/Cas9; 2) To validate the activation of the autonomous interferon signalling in the knock-in cells by Oxaliplatin treatment.
D) Laboratory Location
The lab is located at Life Sciences Building (LSB3-20) at the main campus of Uni of Newcastle.
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PROJECT DETAILS
A) Project Title: Investigation of pathological mechanisms of the schizophrenia risk gene mir137
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
We have shown that ER stress is altered in severe asthma. However, whether ER stress plays a role in, is a consequence of, or can be therapeutically targeted for the treatment of disease, is yet to be fully explored. In this study, students will help conduct a complementary series of clinical (human data, cells and tissues) and experimental (mouse models) investigations to explore how ER stress is affected in the lungs in asthma and how the therapeutic targeting of ER stress, affects disease. This project is funded by the NHMRC Hypothesis: Endoplasmic reticulum (ER) stress plays an important role in the pathogenesis and severity of asthma. The therapeutic targeting of ER stress in asthma with FDA-approved drugs may represent an effective therapeutic for the treatment and/or prevention of disease. Aims: 1.) Define the molecular mechanisms of how ERS/UPR drives bronchial epithelial remodelling, with goblet cell
metaplasia and mucus hypersecretion, and airway fibrosis in severe asthma: We will grow primary bronchial
epithelial cells in air-liquid interface cultures and treat with, chemical ERS inducers, TH2 cytokines and
allergens and investigate molecular signalling pathways using qPCR, PCR arrays,westernblot, Elisa, RNA
sequencing etc.
2. Elucidate the role of ERS in crosstalk between airway epithelium and fibroblasts that drives airway remodelling
and fibrosis in severe asthma: We will establish an epithelial-fibroblast co-culture model and treat this model
with ERS inducers/inhibitors and examine airway remodelling and fibrosis markers using an array of
molecular techniques.
3. Elucidate the mechanistic role of ERS/UPR and therapeutic potential of chemical chaperones in airway
inflammation and remodelling in-vivo in experimental severe steroid-resistant asthma: We will use our
established murine models to see the effect of FDA approved ER stress inhibitors on controlling asthma
symptoms and investigate mechanisms how ER stress contributes to the pathogenesis of asthma.
D) Laboratory Location: Level 2 East and West, Hunter Medical Research Institute
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PROJECT DETAILS
A) Project Title: Investigating the role of hypothalamic populations in motivational deficits following
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Are stress-induced deficits hypothalamic of CRH cell activity associated with impairments in motivation
Stress is a leading risk factor for the development of depression. A debilitating feature of
depression is a loss of motivation. Corticotrophin releasing hormone (CRH) neurons in the
paraventricular nucleus (PVN) of the hypothalamus sit at the apex of the stress hormone
system (the HPA axis). These stress sensitive neurons orchestrate changes in behaviour and
physiology that depend on the release of the stress hormone cortisol (or corticosterone
(CORT) in rodents) from the adrenals. Importantly, recent work has established that CRH
neurons can also control stress related behaviour independent of CORT. Further, when we
used optogenetic stimulation to repeatedly activate CRH neurons, we found that this reduced
motivational behaviour, similar to how repeated/chronic stress can produce motivational
deficits and depression in humans. This project aims to understand how repeated stress
dysregulates the activity of CRH neurons to promote impairments in motivation.
To study CRH cell activity under stress CRH-cre mice will receive injections of the genetically encoded Ca2+ indicator gCaMP6f. Motivation after stress will be assessed using our well-established progressive ratio (PR) prcoedures. In a PR test, animals are first trained to perform an action (typically a lever press) to receive a reward e.g. something palatable like sucrose. After further training, PR testing demands that effort is increased exponentially to continue receiving a reward i.e. the lever pressing requirement to receive sucrose is increased exponentially. Eventually animals will cease to press, a point considered the “break point” which is considered an index of motivation or effort. We hypothesise that CRH cell activity is suppressed by repeated stress which will be associated with impairments in PR responding for sucrose. D) Laboratory Location: MS307 (CAL)
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PROJECT DETAILS
A) Project Title: Investigating the role of hypothalamic populations in motivational deficits following
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Characterisation of a fast acting ventromedial hypothalamus to arcuate nucleus projection that
suppresses feeding.
Until recently, the only satiety promoting neurons known to exist in the brain were thought to be
arcuate nucleus (ARC) proopiomelanocortin (POMC) neurons. Genetic disruption of POMC and its
receptor leads to severe weight gain in mice and humans, although it typically takes 12-24 hours for
optogenetic stimulation of POMC neurons to reduce food intake. However, a novel population of fast-
acting glutamatergic neurons in the ARC that rapidly promotes satiety have been discovered. These
fast-acting ARC Vglut2 neurons are intermingled with POMC neurons in the ARC and rapidly
suppress food intake. This unexpected finding raises the question as to why the brain has evolved
fast and slow populations of satiety-promoting neurons. In the context of acute threat detection, a
rapid satiety response would seem advantageous to promote the shift towards avoidance behaviours.
Thus, we will test the hypothesis that VMH Vglut2 and fast-acting ARCVglut2 are synaptically
connected.
To address this question, we will establish the neural connectivity between VMH Vglut2 and
ARCVglut2 neurons using slice electrophysiology. In order to establish that VMH Vglut2 and
ARCVglut2 neurons are synaptically connected we will use slice electrophysiology in Vglut2 cre::R26-
LSL-tdTomato reporter mice bilaterally injected with AAV-EF1a-DIO-hChR2(H134R)-YFP into
theVMH (n=10). We will record ARCVglut2 td-tomato neurons during photostimulation of VMH
neurons (~50 neurons). In order to show that this circuit is modulated by hunger status, we will record
excitatory postsynaptic currents (EPSCs) on to ARCVglut2 tdtomato neurons in slices collected from
fed (n=10) and fasted mice (n=10). We will measure the intrinsic excitability (voltage-activated
currents), degree of spontaneous action potential (AP) discharge, and the characteristics of evoked
AP discharge using depolarizing step injections and predict greater excitatory inputs onto ARCVglut2
tdtomato neurons in fasted mice.
Outcomes: We predict that photostimulation of VMHVglut2 neurons will excite ARCVglut2 neurons in a monosynaptic manner and decreased excitatory input on to ARCVglut2 tdtomato neurons in the fasted state.
D) Laboratory Location: MS308/MS412 (CAL)
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PROJECT DETAILS
A) Project Title: Deciphering a novel molecular mechanism of therapy resistance in breast cancer.
C) Background and Summary of Proposed Research, including a clear hypothesis, aims and
experimental approach:
Diffuse midline gliomas (DMGs) are highly aggressive brainstem tumours that grow within the structures of brain’s middle axis and are responsible for half of all brain cancer-related deaths in children worldwide. The brainstem controls life-essential functions such as heart rate, swallowing and breathing, meaning a tumour growing in this midline region has devastating consequences. Primarily diagnosed in children, patients and their families face a survival prognosis measured in months– a figure that has not improved despite decades of research.
Two key major hurdles preventing improved outcomes for DMG patients are i) the brain’s highly protective barrier (the ‘blood-brain barrier’ (BBB)) which prevents toxins in the peripheral bloodstream from passing into the brain, and ii) a lack of DMG specific therapies. The BBB precludes traditional chemotherapies from reaching (and selectively killing) DMG tumours, and hence development or enhancement of BBB-penetrable drugs is desperately needed.
PI3K/Akt/mTOR is an intracellular signalling pathway implicated in the growth, survival, and metabolism of almost all cancer types. In collaboration with researcher from the Hudson Institute of Medical Research, we performed CRISPR-Cas9 mediated genome wide deletion of 5,000 genes, revealing numerous genes mapping to the PI3K/Akt/mTOR axis are needed for DMG cell survival. This highlights the importance of simultaneous targeting multiple nodes of the oncopathways. Thanks to Material Transfer Agreements established with each respective company, A/Prof Dun has exclusive access to a suite of promising anti-DMG therapies known to penetrate the BBB: a) Paxalisib (Kazia Therapeutics) an orally-available, small molecule inhibitor of PI3K/Akt. b) “7” is a pan-mTOR inhibitor (Novartis International). iBET858 (GlaxoSmithKline) targets Bromodomain and extraterminal (BET) proteins activated downstream of PI3K/Akt/mTOR.
Therefore, the hypothesis of this study is that CNS optimised therapies targeting oncoproteins required for the survival of DMG will help to improve outcomes.
To provide preclinical confirmation of our hypothesis we will:
Aim 1: Utilise DMG patient-derived cell lines and assess downstream cell signalling following acute treatment with PI3K/mTOR and BRD inhibitors using phosphoproteomics.
Aim 2: Employ DMG PDX models to test the brain pharmacokinetics (PK) and pharmacodynamics (PD) of combinations of PI3K/mTOR and BRD inhibitors.
Aim 3: Assess the survival benefit of optimised combinations PI3K/mTOR and BRD inhibitors using a DMG patient derived xenograft mouse model.
C Background and Summary of Proposed Research, including a clear hypothesis, aims and experimental approach:
Background: Schizophrenia (SZ) is a psychotic disorder that arises from a complex interplay of genetic and environmental
factors. Epidemiological data suggests susceptibility to infections and alteration of inflammatory processes are involved in
SZ, however, the precise role of immunological factors in the disorder remains unclear. C-reactive protein (CRP), a peripheral
biomarker of inflammation, is particularly interesting because it contributes to the acute-phase inflammatory processes
during infection or tissue damage. While observational studies have identified elevation of serum CRP in individuals with SZ,
suggesting a potentially causal role mediated via aberrant inflammation, recent genetic analyses from our group suggests
elevated serum CRP exhibits a protective effect on SZ. Paradoxically, we also observed it to be associated with cortical
thinning in brain regions affected by the disorder. We therefore suspect these discordant findings may stem from complex,
recently discovered anti-inflammatory functions of CRP, however further investigation is required to further characterise
and explore the relationship between CRP, SZ, host responses to infection and general neuronal function.
Hypothesis and aims: We hypothesise that genetic risk for SZ is associated with CRP signalling, and these same genetic risk
factors also influence host-responses to infection. To address this, the following aims will be investigated:
1. To explore the relationship between SZ genetic risk, host antigen responses and CRP.
2. To experimentally characterise the implications of CRP dysregulation on neuronal morphology, integrity and gene
expression in vitro.
Research plan: The project will examine the relationship between SZ genetic risk and host IgG titres for 14 pathogens utilising
the UK Biobank cohort, consisting of over 500,000 individuals with matched genetic and serological data. The effect of
adjusting for measured and/or genetically proxied CRP on the association between antigen response and SZ genetic risk will
be quantified to establish whether it impacts this relationship.
The effect of CRP on neuronal properties will then be explored in cultured neurons via manipulation CRP concentrations. The
resultant impact on neuronal morphology, function, integrity and gene expression will be characterised by live cell
microscopy, multielectrode array, viability assays and total RNA sequencing, respectively.
D Laboratory Location MSB613
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LIST OF APPROVED PROJECTS in alphabetical order by primary supervisor (have a student involved as of 8/11/21) Project Title: Examining endocannabinoid signalling in the gut-brain-axis in an animal model of
adolescent cannabinoid exposure
Supervisors: Dr Grace Burns, Prof Deborah Hodgson
Project Title: Understanding the gut-brain axis in pre-term babies and neonatal colitis
Supervisors: Dr Bridie Goggins, Dr Peter Pockney
Project Title: Investigating the effects of bushfire smoke exposure on maternal health, and
respiratory health of offspring
Supervisors: A/Prof Jay Horvat, Dr Alexandra Brown
Project Title: Developing a 3D-printed ‘gut-on-a-chip’ platform to study the gut-brain axis
Supervisors: Dr Gerard Kaiko, Prof Simon Keely
Project Title: Hypoxic signalling and the link between intestinal inflammation and colorectal cancer
Supervisors: Prof Simon Keely, Dr Emily Hoedt, Dr Grace Burns
Project Title: Investigation of a novel multipotent inflammasome inhibitor for the treatment of
influenza A virus infections
Supervisors: Dr Jemma Mayall, A/Prof Jay Horvat
Project Title: Role and therapeutic targeting of iron responses in influenza A virus infections
Supervisors: Dr Jemma Mayall, Dr Alexandra Brown
Project Title: Regulation of s(P)RR release from the placenta
Supervisors: A/Prof Kirsty Pringle, Prof Eugenie Lumbers, Dr Sarah Delforce, Dr Saije Morosin
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Project Title: Unfurling the mechanisms of (P)RR driven placental development
Supervisors: A/Prof Kirsty Pringle, Prof Eugenie Lumbers, Dr Sarah Delforce, Dr Saije Morosin
Project Title: Characterising a new potential regulator of AML cell invasion
Supervisors: A/Prof Kathryn Skelding, A/Prof Lisa Lincz
Project Title: Examination of a Ca2+ modulator as a potential treatment for AML
Supervisors: A/Prof Kathryn Skelding, A/Prof Lisa Lincz
Project Title: Understanding the underpinnings of the declines in cholesterol and protein
homeostasis in the aging CNS
Supervisors: A/Prof Doug Smith, Dr Mitchell Cummins
Project Title: Fertility and sexual health knowledge and priorities in young people
Supervisors: Dr Jessie Sutherland, Dr Emmalee Ford
Project Title: The role of immune-cell derived exosomes in chlamydia infection and fertility
Supervisors: Dr Jessie Sutherland, Prof Eileen McLaughlin, Dr Elizabeth Bromfield
Project Title: Neuroimmune interactions associated with neuropsychiatric disorders following early
life inflammation
Supervisors: Dr Melissa Tadros, Dr Lauren Harms
Project Title: Development of Dried Blood Spot (DBS) Assays for Tyrosine Kinase Inhibitors
Supervisors: A/Prof Paul Tooney, A/Prof Jenny Schneider, Dr Peter Galettis
Project Title: Preclinical testing of novel combination therapies for acute myeloid leukaemia
Supervisors: A/Prof Nikki Verrills, Dr Heather Murray, Dr Joshua Brzozowski