Cancer
Glioma stem cells: biology and molecular targets
Supervisor: Dr Andrew Morokoff, Dr Kate Drummnod, Dr Giovanna D’Abaco
Location: Department of Surgery, Royal Melbourne Hospital
Contact: Dr Andrew Morokoff (morokoff@unimelb.edu.au) 9342 7703
Gliomas are highly invasive brain tumours with an extremely poor survival because of their highly invasive nature and high recurrence rate. Recently, a subpopulation of cells (CD133+) with stem cell-like properties have been identified in gliomas and these cells are thought to be the primary cause of recurrence and treatment resistance. Furthermore, certain molecular pathways that lead to invasion, anti-apoptotic and drug resistance effects may be ‘switched on’ in glioma stem cells. Thus, understanding this type of cell may lead to better treatments. Thus project involves establishing stem cell cultures from surgical brain tumour specimens and growing then in special conditions as ‘neurospheres’. Once the cell lines are established they will be assessed for know alterations of molecular signalling pathways and genetic mutations. This data will be collated and compard to clinical data from the corresponding patients such as time to progression and survival to form a ‘genetic signature’ of these tumours. This information will form the underpinning to the testing of various inhibitors against targets such as PI3K in glioma stem cells.
Circulating endothelial cells as a biomarker in brain tumours
Supervisor: Dr Andrew Morokoff, Dr Chris Hovens, Dr Kate Drummond
Location: Department of Surgery, Royal Melbourne Hospital
Contact: Dr Andrew Morokoff (morokoff@unimelb.edu.au) T: 9342 7703
There has been much interest recently in defining better markers of brain tumour behaviour and prognosis. At the present time, prognosis is based on histopathology grading scheme (WHO I-IV 2007) and the response of the tumour to treatment is followed on contrast-enhanced MRI. However, both of these methods have limitations, particularly in the context of post-treatment effects after both radiation therapy and especially novel chemotherapies that target angiogenesis. Endothelial cells that line tumour blood vessels apoptose and enter the blood stream, becoming detectable as Circulating Endothelial Cells (CECs) with a simple blood test. Levels of CECs have been shown to correlate with the activity of the tumour in prostate cancer, and therefore hold great promise for brain tumours, which are highly angiogenic. The project involves taking blood samples from brain tumour patients and testing for the levels of CECs by using FACS analysis. This identifies specific markers of endothelial cells such as CD31, CD45 and VEGFR2. By taking samples pre and post treatment time points, we hope to correlate the CEC level with the tumour burden and determine if this could provide more information as a clinically relevant biomarker in brain tumours.
Dynamin as an anti-tumour drug in gliomas
Supervisor: Dr Andrew Morokoff, Prof Terence O’Brien, Prof Phil Robinson
Location: Department of Surgery and Medicine, RMH. Department of Physiology, Children’s Medical Research Institute, Sydney.
Contact: Dr Andrew Morokoff T: 9342 7703 E: (morokoff@unimelb.edu.au)
Prof Terence O’Brien E: (tjobrien@unimelb.edu.au)
Background: The group of Prof Phil Robinson at CMRI have developed dynamin inhibitors as promising treatments of epilepsy and possibly also gliomas. Gliomas are highly invasive and recurrent tumours that have a particularly poor prognosis, despite surgery, radiotherapy and chemotherapy. Novel effective treatments for glioma are desperately needed. A number of inhibitors of dynamin II (dyn-II) have been screened for anti-tumour activity against glioma cell lines and have shown positive effects.. This project aims to test these inhibitors in an in vivo animal model of gliomas. Glioma cells will be injected stereotactically into the amygdala of 8-12 week old nude mice and the animals asssessed or tumour growth and survival after treatment with or without study drug. The tumour growth will be assessed both by bioluminescence with live animal imaging (IVIS system) as well as be immuohistochemistry and volumetric brain slice analysis. The student will develop skills with animal handling, cell lines, in vivo imaging and data/statistical analysis.
TGF-signalling and cancer development
Supervisors: Dr. Hong-Jian Zhu (and Dr. Rodney Luwor, Bo Wang, Catherine Winbanks)
Location: Cancer Signalling Laboratory, Department of Surgery (RMH) (5th Floor, Clinical Sciences Building, The Royal Melbourne Hospital)
Contact : Dr Hong-Jian Zhu T: 8344 3025 Email hongjian@unimelb.edu.au
Project Outline: Traditionally, key-lock or on-off models dominate the molecular understanding of cellular signalling and disease development, with most studies focusing on linear molecular signalling cascades. With the advent of large scale molecular techniques such as proteomics and microarrays, cross-talk between signalling networks has been implicated to play critical roles in cancer development. It challenges the physiological validity of the switch on-off model. Our lab, using molecular, cellular and gene targeted animal models as well as human patient samples, has established that the moderation of signalling sensitivity by other pathways, rather than a black-white switch on-off, specifically of the TGF- (Transforming Growth Factor-) signalling pathways determines cancer progression. These findings have been published in top-ranking biomedical journals including Nature Medicine (11:845-52, 2005). Given the medical significance, current works in our lab are supported by 4 NHMRC and 1 Cancer Council grants totalling more than $2 million.
This lab aims to understand the molecular fundamentals of TGF- signalling mis-regulation and its causation effect on early tumor development and late tumor invasion and metastasis. In particular, we focus on the few major oncogenic molecular pathways’ cross-talk with TGF- signalling in various stages and types of cancer development. Concurrently, we are also devising strategies utilizing our unique molecular insights to convert tumor-causing signalling to directly tumor-killing.
The following projects are designed for students to participate in forefront cancer research and to achieve excellent novel results in a relative short time frame (9-10 months).
Project A: Converting oncogene signalling to tumor killing in brain cancer
Project B: Stat3 mediated impairment of TGF- signalling in head&neck and breast cancer
Project C: Targeting TGF- signalling expansion in brain tumor invasion
Project D: Regulation of TGF- signaling by Wnt pathway in the development of colon cancer
Techniques to be used: Cell culture, reporter assays (gene expression), adenoviral work, molecular biology, Western and Northern blotting (protein and mRNA respectively), thymidine assays (cell proliferation), real-time PCR, immunofluorescence and immunohistochemistry, siRNA (gene silencing), animal imaging.
Preferred background and quality of student: biochemistry, pathology, medical sciences; good nature as a person, passionate and dedication in research, perseverance in problem solving.
Circulating endothelial cells as biomarkes for prostate cancer
Supervisor: Dr. Chris Hovens
Location: Department of Surgery (RMH) (5th Floor Clinical Sciences Building and Prostate cancer Epworth Hospital, Richmond)
Contact: Dr Chris Hovens T: 9342 7703/4 E : chovens@unimelb.edu.au
The development of a vascular network (angiogenesis) is essential for all solid tumours. Numerous studies have underscored the importance of angiogenesis in the development and progression of prostate cancer. The significant contribution of bone marrow progenitor cells to the vascularisation of a number of different tumour types has recently been recognised. Following angiogenic stimuli, a pool of haematopoietic progenitor cells can become mobilized and contribute to the vascularization and growth of certain primary tumours. These cells are detectable in the circulation as Circulating Endothelial Cells. Significantly, it has recently been shown that these same cells are crucial for setting up a pre-metastatic niche at distinct organ sites where tumour metastasis is prevalent. We propose to determine whether measuring bone marrow endothelial cell recruitment to tumours may be of benefit in stratifying the risks of progression and metastases in patients with prostate cancer, and possible response to treatment.
Benefits to student: Molecular and clinical research in the one project, multi-collaborative project encompassing basic research and clinical interaction
Requirements for students: Dedicated, passionate and committed. Must have done well academically.
Genetic and pharmacologic approaches to dissect lung inflammation and lung cancerSupervisors: A/Prof Margaret Hibbs (Ludwig Institute) and Professor Gary Anderson (Department of Pharmacology, University of Melbourne)
Location: Department of Pharmacology, University of Melbouorne
Contacts: Professor Gary Anderson T: 8344 8602 E: gpa@unimelb.edu.au
A/Professor Margaret Hibbs T: 9341 3155 E: Margaret.Hibbs@ludwig.edu.au
Chronic obstructive pulmonary disease (COPD) is an incurable and often fatal inflammatory lung disease, and is a known risk factor for lung cancer. We have a number of animal models of inflammatory lung disease, including mice with activating mutations in Src family kinases, and mice with deleterious mutations in the inositol phosphatase SHIP-1 or the protein tyrosine phosphatase SHP-1. The aim of this project is to use genetic approaches to identify genes that predispose to inflammatory lung disease, and pharmacologic methods to reverse establish disease.
Skill acquisition: In vivo disease models, quantitative PCR, cell culture, histology, FACS analysis of cell populations; immuno-affinity purification of proteins, immune regulation and transduction biochemistry.
Src kinases, lung inflammation and lung cancer
Supervisors: A/Prof Margaret Hibbs and Professor Gary Anderson
Location: Department of Pharmacology, University of Melbourne
Contacts: Professor Gary Anderson T: 8344 8602 E: gpa@unimelb.edu.au
A/Professor Margaret Hibbs T: 9341 3155 E: Margaret.Hibbs@ludwig.edu.au
Lung cancer is now the most common cause of cancer death in the world. We have discovered that mutations in src kinases cause lung cancer even though the mutated kinases are not themselves expressed in lung tissue. Dys-regulated inflammation seems to be the underlying problem. This project will study exactly how inflammation causes lung cancer.
Skill acquisition: In vivo disease models, quantitative PCR, cell and tissue culture, histology, FACS analysis of cell populations; immuno-affinity purification of proteins, immune regulation and transduction biochemistry.
The role of Wnt/b-catenin and Stat3 signalling in cancer
Supervisors: Dr Michael Buchert, Dr Toby Phesse (Ludwig Institute)
Location: Ludwig Institute for Cancer Research
Contact: T : 03 9341 3155 Dr Michael Buchert E : michael.buchert@ludwig.edu.au Dr Toby Phesse E: toby.phesse@ludwig.edu.au
The canonical Wnt and the cytokine-activated Stat3 signalling pathways are key drivers for tumourigensis in a variety of human tumours. In our laboratory, we have developed genetically modified mice in which both the Wnt and Stat3 signalling pathways are activated in the gastrointestinal tract. This results in the formation of tumours in the small and large intestine as well as in the stomach. In addition we are setting up a mouse model in which skin tumours can be induced chemically in mice with activated Wnt and Stat3 signalling pathways. The aim of this project is to elucidate the mechanism(s) leading to tumour formation with a focus on identifying potential avenues for reversing tumour growth/formation.
Skill acquisition: The successful BSc honours student will be using a combination of molecular and biochemical techniques such as quantitative real-time PCR, histology, immuno-histochemistry, Western blotting, cell culture etc on biological samples derived from genetically engineered mice.
Role of the transcription factor, c-Myb in cell growth and proliferation in the vertebrate intestinal epithelium
Supervisors: A/Professor Joan Heath, A/Professor Rob Ramsay
Location: Ludwig Institute for Cancer Research
Contacts:
Associate Professor Joan Heath:
Joint-head, Colon Molecular and Cell Biology Lab,
Ludwig Institute for Cancer Research, Parkville Campus.
T: 9341 3150.
Email: joan.heath@ludwig.edu.au
Assoc Professor Rob Ramsay:
Peter MacCallum Cancer Centre, Research Division, East Melbourne. T: 9656 1863. Email: rob.ramsay@petermac.org
The highly elaborate epithelial lining of the vertebrate intestine is a dynamic and self-renewing tissue system that encompasses most aspects of cell behaviour, including cell proliferation, differentiation, migration and apoptosis. To a large extent, the genetic mechanisms involved in establishing and maintaining this constantly remodelling tissue system remain a mystery. Due to its many favourable characteristics, including prolific reproduction, external development and optical transparency of embryos, the zebrafish is an ideal model for the genetic analysis of vertebrate organogenesis.
In the zebrafish intestine, three distinct cell lineages are derived from a common multipotential stem cell. These cells undergo a series of binary cell fate decisions to give rise to the enterocytes (nutrient absorbing), goblet (mucous producing) and enteroendocrine (hormone producing) cells. The mechanisms that govern these binary cell fate decisions are incompletely understood. We recently identified a new BAC transgenic line, Tg[c-myb:YFP], which provides an exciting opportunity to throw light on this question. In this line, the regulatory elements of the c-myb gene drive strong YFP expression in an abundant population of cells scattered throughout the intestinal epithelium.
The specific aim of this project is to characterize the genetic regulation of epithelial cell growth and differentiation in the zebrafish intestine using reverse genetic approaches. Specifically, antisense morpholino oligonucleotides, targeted to c-myb, (a transcription factor known to play a role in intestinal development) will be injected into the yolk of 1-2 cell zebrafish embryos in order to knock-down c-Myb function over the first few days of development. The impact of inhibiting this transcription factor on intestinal epithelial cell development will be analysed in the first instance using fluorescence dissecting and confocal microscopy. Other approaches will be to examine intestinal epithelial cell development in a panel of zebrafish intestinal mutants that are currently undergoing characterization in our laboratory using positional cloning, in situ hybridization and immunohistochemistry. This analysis will be greatly facilitated by establishing the mutant strains onto the transgenic Tg[c-myb:YGF] background.
This Hons project will largely be conducted in the Colon Molecular and Cellular Biology Laboratory, Ludwig Institute for Cancer Research, Royal Parade, Parkville, where all the facilities for zebrafish developmental genetic studies are located.
Analysis of the APC tumour suppressor protein in 3D cell culture models
Supervisors: Dr Maree Faux, Professor Tony Burges, Ludwig Institute for Cancer Research
Location: Ludwig Institute for Cancer Research
Contact: Dr Maree Faux T : 03 9341 3155 Email : Maree.faux@ludwig.edu.au
Colon cancer is one of the major diseases of the Western world and affects more people in Australia than any other cancer. APC mutations can be inherited, but more than 80% of sporadic colon cancers carry truncating mutations in the tumour suppressor protein APC (adenomatous polyposis coli). APC mutations are thought to be an early event in a multistep process involving the successive acquisition of genetic mutations. This suggests a key role for Apc in the maintenance of normal colonic cellular function, however, the precise mechanism of events arising from its loss of function that lead to the development of polyps and adenomas is not known. A well established role for APC is in the regulation of the Wnt signaling target b-catenin. Recent studies demonstrate that APC is also involved in cytoskeletal regulation and is likely to play a role in cell migration, adhesion and differentiation. We have developed antibodies, recombinant proteins and cell lines for the study of different aspects of APC structure and function. We have evidence that the wild-type protein can influence cell adhesion. We believe that APC is a key mobile scaffold regulating cell adhesion and that its functions are intimately linked with its location and dynamic behaviour in the cell. The aim of this project is to investigate mutated and wt APC at the subcellular level in 3D culture models. The human colorectal cancer cell line LIM1863, containing mutated APC protein, form three-dimensional highly organised, multicellular structure organoids that resemble enclosed carcinoma tubules. Polarized MDCK epithelial cells, containing full-length APC, form three-dimensional cysts in culture. These models will be used to assess endogenous APC, as well as organoid/cyst formation in cells expressing APC-GFP and APC siRNA.
Skill acquisition: The successful BSc honours student will use a combination of molecular and biochemical techniques such as cell culture, immunostaining, confocal microscopy, transfection, and Western blot analysis.
Characterization of the role of Th17 cell populations in gastrointestinal cancer.
Supervisors: Dr Tracy Putoczki, A/Professor Matthias Ernst
Location: Ludwig Institute for Cancer Research
Contact: Dr Tracy Putoczki T: 9341 3155 Email : Tracy.Putoczki@Ludwig.edu.au
A/Prof Matthias Ernst T: 9341 3155 Email: Matthias.Ernst@Ludwig.edu.au
Project (including aims): Recently, the classical T helper-cell paradigm was challenged by the discovery of a new T-helper cell lineage, coined Th17. These cells have been implicated in a growing list of autoimmune disorders including psoriasis, arthritis, and multiple sclerosis and most recently they have been associated with cancer development. In contrast, regulatory T-cells (Tregs) are involved in managing appropriate immune responses to pathogen invasion and tissue damage. The role of this cell population in inflammation-associated cancer progression is not well understood. This project will explore the contribution of Th17 and Treg cell populations to gastrointestinal cancer development. We have a number of animal models of inflammation-associated gastrointestinal cancer which will be used in conjunction with a range of cellular biology methods to understand how these cells participate in the inflammation associated with cancer.
Skill Acquisition: In vivo disease models, analysis of genetic knock-in and knock-out mouse strains, histology, quantitative PCR, cell culture, FACs analysis, Elisa, Western blotting.
Using a new mouse model to understand colitis.
Supervisors: Dr Tracy Putoczki, A/Professor Matthias Ernst
Location: Ludwig Institute for Cancer Research
Contact: Dr Tracy Putoczki T: 9341 3155 Email : Tracy.Putoczki@Ludwig.edu.au
A/Prof Matthias Ernst T: 9341 3155 Email: Matthias.Ernst@Ludwig.edu.au
Project (including aims): We have generated a novel transgenic mouse model in which a molecule called signal transducer and activator of transcription (Stat3), which utilizes gp130 receptor signalling, is constitutively expressed in a tissue specific and ligand independent manner. Stat3 has been demonstrated to provide a tissue protective function in inflammatory bowel disease (IBD), such as acute colitis, through activation of downstream target genes. However in a situation of chronic inflammation, Stat3 is associated with colon cancer development. The balance of Stat3 signalling required to be beneficial or deleterious for these diseases is not understood. In the first instance, this project will review the functionality of the DNA constructs used to generate the mouse model. In addition, this project will utilize the novel transgenic mouse descrived in a variety of models of IBD to fully characterize and further understand the role of Stat3 in colitis. Visualization of disease will be aided by the use compound mutant mice in which the transgenic mouse is crossed with a mouse expressing a luciferase reporter construct that will allow for in vivo imaging of the colonic epithelium using state of the equipment.
Skill Acquisition: In vivo disease models, in vivo animal imaging, analysis of transgenic and genetic knock-in and knock-out mouse strains, histology, quantitative PCR, Western blotting, molecular biology including vector design and recombinant DNA techniques.
What role do T-cells play in colitis?
Supervisors: Dr Tracy Putoczki, A/Professor Matthias Ernst
Location: Ludwig Institute for Cancer Research
Contact: Dr Tracy Putoczki T: 9341 3155 Email : Tracy.Putoczki@Ludwig.edu.au
A/Prof Matthias Ernst T: 9341 3155 Email: Matthias.Ernst@Ludwig.edu.au
Project (including aims): Individuals affected by chronic inflammatory diseases such as inflammatory bowel disease (IBD) are highly susceptible to developing colonic cancers. IBD refers to chronic diseases that cause inflammation of the intestine: ulcerative colitis (UC) and Crohn’s disease (CD). These diseases affect approximately 1% of Australians and lead to significant pain and discomfort for which there is no current cure. This project will utililze a mouse model for CD, referred to as the CD45 T-cell transfer model to establish the role different T-cells populations play in colitis and ultimately the role they may play in cancer development. The project will take advantage of a number of established knock-in and knock-out mouse models for numerous genes involved in T-cell development that also have suspected roles in cancer.
Skill Acquisition: In vivo disease models,analysis of transgenic and genetic knock-in and knock-out mouse strains, histology, quantitative PCR, cell culture, FACs analysis and Western blotting.
The role of PTEN and Stat3 signalling in cancer
Supervisors: Dr Michael Buchert, Dr Toby Phesse (Ludwig Institute)
Location: Ludwig Institute
Contact: T: 03 9341 3155 Dr Michael Buchert E : michael.buchert@ludwig.edu.au
Dr Toby Phesse E: toby.phesse@ludwig.edu.au
The tumour suppressor PTEN is one of the most commonly mutated genes in human cancer and the cytokine-activated Stat3 signalling pathways are key drivers for a wide range of human pathologies, most notably cancer. In our laboratory, we have developed genetically modified mice which carry mutations that inactivate PTEN and hyper-activate the Stat3 signalling pathway. This results in the formation of various tumours affecting different tissues, among them the gastrointestinal tract. The aim of this project is to elucidate the mechanism(s) leading to increased tumour formation in PTEN/Stat3 mutated mice with a focus on identifying potential avenues for reversing tumour growth/formation.
Skill acquisition: The successful BSc honours student will be using a combination of molecular and biochemical techniques such as quantitative real-time PCR, histology, immuno-histochemistry, Western blotting, cell culture etc on biological samples derived from genetically engineered mice.
Exploiting non-oncogene addiction for therapeutic purposes in a preclinical mouse model of gastric tumourigenesis.
Supervisors: A/Professor Matthias Ernst, Dr Tracy Putoczki
Location: Ludwig Institute for Cancer Research
Contact: A/Prof Matthias Ernst T: 9341 3155 E: Matthias.Ernst@Ludwig.edu.au
Dr Tracy Putoczki T: 9341 3155 E : Tracy.Putoczki@Ludwig.edu.au
Project: Cancers of the gastrointestinal tract are often associated with chronic inflammation and represent a major health burden. These malignancies commonly show aberrant activation of the latent transcription factor Stat3 that promotes proliferation, cell survival and angiogenesis. Our previously developed the gp130F/F knockin mutant mouse provides a clinically relevant, fully penetrant preclinical mouse model for inflammation-associated intestinal-type gastric cancer, in which neoplastic disease shares many histological hallmarks with the human malignancies and is dependent on interleukin-6 cytokine family-mediated Stat3 hyperactivation. While therapeutic interference with this signaling axis shows some beneficial effect on tumour burden in gp130F/F mice, this project takes advantage of an emerging finding that non-mutated proteins and their associated pathways, often become rate limiting for neoplastic growth (referred to as “non-oncogene addiction”). This project will test the efficacy of pre-clinical drugs to target such pathways and explore whether they provide potential therapeutic value for the treatment of Stat3- and/or inflammation-dependent solid tumours.
References: Jenkins B, Ernst M et.al., (2005) Nature Medicine; Ernst et.al., (2008) J Clin Invest; Bollrath J, Putoczki T, Ernst M et.al (2009) Cancer Cell
Skill Acquisition: In vivo disease models, analysis of genetic knock-in and knock-out mouse strains, histology, quantitative PCR, cell culture, FACs analysis, Elisa, Western blotting.
Regulation of Stat3-mediated Tumor Progression
Supervisors: Dr Rodney Luwor
Location: Department of Surgery, Royal Melbourne Hospital
Contact: T : 9342-7703 E : rluwor@unimelb.edu.au
During physiological processes the intracellular protein Signal Transducer and Activator of Transcription 3 (Stat3) is activated by many growth factors and cytokines (e.g. EGF) resulting in entry into the nucleus and transcription of many genes involved in a multitude of cellular processes. However, uncontrolled or un-attenuated stat3 phosphorylation/activation results in cancer initiation, progression and metastasis of many tumour types. Therefore, understanding how stat3 is regulated or controlled within the cell is pivotal for cancer biology and may allow greater scope for therapeutic intervention into stat3-driven tumourigenesis. Recently, we showed that EGFR-driven stat3 phosphorylation and transcriptional activity is regulated by EGFR internalisation indicating that EGFR activates stat3 while it traffics’ through the cell in the endosomal pathway. Therefore this project aims tolocate where the EGFR phosphorylates Stat3 within the cell utilizing a panel of siRNA targeting proteins essential for EGFR trafficking.
Skills acquisition: Cell biology techniques including Cell transfections, western blotting, immunofluorescence staining and confocal microscopy, luciferase reporter assays and potentially animal handling and injecting.