Arthritis and Inflammation Research
Arthritis and Inflammation Research Centre
The Arthritis and Inflammation Centre is headed by Professor John Hamilton who leads a team of scientists that focuses on inflammation-associated diseases, including arthritis, host pathogen interaction and cancer. The pathology of most diseases involve some degree of inflammation with macrophages often being the major cell type; as a result the Centre focuses primarily on macrophage biology and the effects of macrophage-associated inflammation on other cell types such as stem cells.
We employ a variety of techniques and strategies including gene-based strategies (for example, micro-array technology) to understand disease causation, protein-based strategies (including proteomics, immunoprecipitation, cell transfection) to study the cellular signal transduction pathways associated with disease, and mouse models and clinical material to analyse disease in vivo.
Key components of the biology involve an analysis of how macrophage lineage cells are altered during inflammatory disease, how at a molecular level these cells survive, proliferate, differentiate or are activated, and how to down-regulate the cellular functions aberrant in disease. There is some emphasis on growth factor biology/biochemistry and on signal transduction pathways implicated strongly in human arthritis, cancer and stem cell biology.
The role of urokinase plasminogen activator (u-PA) and its receptor (u-PAR) in arthritis and inflammation
Supervisor: Dr Andrew Cook
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Andrew Cook T: 8344 3290 Email: adcook@unimelb.edu.au
Rheumatoid arthritis (RA) is a chronic inflammatory disease affecting approximately 1% of the population. Fibrin deposition, cell migration, and tissue destruction and remodeling are key components in the pathology of RA joints. The plasminogen activators (PAs), urokinase (u-PA) and tissue-type (t-PA), which converts plasminogen to plasmin, are implicated in these processes; however their precise roles in such processes, particularly for u-PA and its receptor (u-PAR), have yet to be defined. In this project you will study the role of u-PA and the u-PAR, in inflammation and arthritis using mice genetically altered mice such that u-PA or u-PAR have been rendered inactive. In particular, the effect of u-PA on cell migration to an inflammatory site, on tissue destruction and remodeling, and in activating/suppressing other key cytokines/proteases (eg metalloproteinases (MMPs)) involved in these processes will be studied.
Skill acquisition: experience with animal models of human disease, measurement of inflammatory mediator mRNAs by real time-PCR and their concentrations by ELISA, and the use of FACS and immunohistochemistry to study cell populations.
The role of granulocyte macrophage colony stimulating factor (GM-CSF) in arthritis and inflammation
Supervisor: Dr Andrew Cook
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Andrew Cook T: 8344 3290 Email: adcook@unimelb.edu.au
Rheumatoid arthritis (RA) is a chronic inflammatory disease affecting approximately 1% of the population. We have shown that GM-CSF is important for the development of several models of inflammation and arthritis. Furthermore, blockade of GM-CSF is effective at reducing arthritis severity. Phase 1 clinical trials are now underway in human rheumatoid arthritis. However, we still do not completely understand how GM-CSF is acting during inflammation and arthritis. In this project you will study the role of GM-CSF in inflammation and arthritis, and in particular, its role in monocyte/macrophage survival and activation.
Skill acquisition: experience with animal models of human disease, measurement of inflammatory mediator mRNAs by real time-PCR and their concentrations by ELISA, and the use of FACS and immunohistochemistry to study cell populations.
The role of inflammation in mesenchymal stem cell differentiation
Supervisor: Dr Derek Lacey and Prof John Hamilton
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Derek Lacey T: 8344 3292 Email: dlacey@unimelb.edu.au
Mesenchymal stem cells (MSC) have been shown to differentiate into osteoblasts, adipocytes, myocytes and aid in the tissues repair. In the context of chronic inflammatory conditions like rheumatoid arthritis, chronic obstructive pulmonary disease and crohn’s disease, MSC are unable to repair their target tissue for unknown reasons. In this study we propose to determine the mechanisms by which MSC are prevented from undergoing differentiation and tissue repair in the presence of inflammation. Specifically, the project will be examining the signalling pathways involved in blocking MSC differentiation into osteoblasts in the presence of inflammatory mediators. In this project you will be isolating adult mesenchymal stem cells from mice and using a stem cell line to determine the effects of inflammation on stem cell biology.
Skill acquisition: a variety of molecular and cell biological, and biochemical techniques, such as PCR and cloning of recombinant DNA; tissue culture, and FACS analysis; immuno-affinity purification of proteins, SDS-PAGE and Western blotting
The role of Wnts in Arthritis
Supervisor: Dr Derek Lacey, Dr Andrew Cook and Prof John.Hamilton
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Derek Lacey T: 8344 3292 Email: dlacey@unimelb.edu.au
Wnts are a family of proteins important in development. Through a microarray screen of macrophage populations we have also found that Wnts are expressed by inflammatory macrophages. Macrophages are key cells involved in the destruction joints during rheumatoid arthritis. This project will investigate the expression of Wnts in patient’s tissue samples and in an inflammatory model of arthritis and determine if targeting Wnts would be a beneficial treatment for arthritis. In this project you will be cutting tissue sections and measuring the expression of Wnts. You will be inducing an murine model of arthritis and measuring a number of clinical parameters and collecting and processing tissue and measuring Wnt expression by histology, real-time PCR, western blotting and FACS analysis.
Skill acquisition: a variety of molecular and cell biological, and biochemical techniques, such as PCR and cloning of recombinant DNA; tissue culture, and FACS analysis, SDS-PAGE and Western blotting
The role of a novel therapeutic target in Arthritis
Supervisor: Dr Derek Lacey, Dr Andrew Cook and Prof John Hamilton
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Derek Lacey T: 8344 3292 Email: dlacey@unimelb.edu.au
Through a proteomic screen of inflammatory macrophages we have identified a novel potential therapeutic target for the treatment of arthritis. Macrophages are key cells involved in the destruction joints during rheumatoid arthritis. This project will investigate the expression of this target in patient’s tissue samples and in an inflammatory model of arthritis and determine if targeting this protein would be a beneficial treatment for arthritis. In this project you will be cutting tissue sections and measuring the expression of this novel protein. You will be inducing a murine model of arthritis and measuring a number of clinical parameters and collecting and processing tissue and measuring protein expression by histology, real-time PCR, western blotting and FACS analysis.
Skill acquisition: a variety of molecular and cell biological, and biochemical techniques, such as PCR and cloning of recombinant DNA; tissue culture, and FACS analysis, SDS-PAGE and Western blotting
The role of Wnts in Macrophages
Supervisor: Dr Derek Lacey, Dr Glen Scholz and Prof John Hamilton
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Derek Lacey T: 8344 3292 Email: dlacey@unimelb.edu.au
Wnts are a family of proteins important in development. Through a microarray screen of macrophage populations we have also found that Wnts are expressed by inflammatory macrophages. Macrophages are key cells involved in the destruction joints during rheumatoid arthritis. This project will investigate the expression of Wnts in macrophages under various inflammatory conditions. You will also overexpress Wnts in a macrophage cell line to determine its role in macrophage function. In this project you will be culturing cell lines and primary cells and measuring the expression of Wnts. You will be cloning a Wnt protein and transfecting cell lines and measuring Wnt expression by histology, real-time PCR, western blotting and FACS analysis.
Skill acquisition: a variety of molecular and cell biological, and biochemical techniques, such as PCR and cloning of recombinant DNA; tissue culture, and FACS analysis, SDS-PAGE and Western blotting
The role of a novel therapeutic target in Macrophages
Supervisor: Dr Derek Lacey, Dr Glen Scholz and Prof John Hamilton
Location: Arthritis Research and Inflammation Centre, Department of Medicine (RMH/WH), University of Melbourne
Contact: Dr Derek Lacey T: 8344 3292 Email: dlacey@unimelb.edu.au
Through a proteomic screen of inflammatory macrophages we have identified a novel potential therapeutic target for the treatment of arthritis. Macrophages are key cells involved in the destruction joints during rheumatoid arthritis. This project will investigate the expression of this novel protein in macrophages under various inflammatory conditions. You will also overexpress this protein in a macrophage cell line to determine its role in macrophage function. In this project you will be culturing cell lines and primary cells and measuring the expression of this protein. You will be cloning this novel therapeutic target protein and transfecting cell lines and measuring its expression by histology, real-time PCR, western blotting and FACS analysis.
Skill acquisition: a variety of molecular and cell biological, and biochemical techniques, such as PCR and cloning of recombinant DNA; tissue culture, and FACS analysis, SDS-PAGE and Western blotting
Host pathogen interactions and mucosal inflammation
Supervisor: Dr Glen Scholz
Location: Department of Medicine (RMH)
Contact Tel: 8344-3298; Email: glenms@unimelb.edu.au
Our respiratory and gastrointestinal tracts represent major portals for the entry of important pathogens (e.g. Influenza virus, H. pylori and Salmonella). The epithelial cells lining mucosal surfaces play a pivotal role in host defence as they express pathogen recognition receptors (e.g. Toll-like receptors) that allow them to initiate an inflammatory response upon infection. Although the inflammatory factors released by epithelial cells recruit and activate leukocytes to deal with the infection, they also contribute to the pathology of chronic inflammation (e.g. Crohn’s disease, ulcerative colitis, and asthma) and cancer (e.g. gastric cancer). In this project you will investigate how a specific member of the Interferon Regulatory Factor (IRF) family of transcription factors uniquely regulates the inflammatory response of epithelial cells to pathogens. This will involve identifying the specific genes that are regulated by the transcription factor, as well as working out how the activity of the transcription factor is regulated in response to specific pathogens.
Skill acquisition: a variety of cell biology (e.g. tissue culture, ELISA assays, FACS analysis, and confocal microscopy), biochemical (e.g. affinity purification of proteins and Western blotting), and molecular biology techniques (e.g. Real-Time PCR, cloning, transfection of cells, and siRNA-mediated gene silencing).
How macrophages use SNAREs to capture and kill microbial pathogens
Supervisor: Dr Glen Scholz
Location: Department of Medicine (RMH)
Contact: 8344-3298; Email: glenms@unimelb.edu.au
The phagocytosis and killing of pathogens by macrophages, as well as their secretion of inflammatory cytokines (e.g. TNF), is crucial for effective host defense. These key immune functions of macrophages depend upon the highly coordinated intracellular trafficking of vesicles from one compartment to another (e.g. trafficking of phagocytosed pathogens to lysosomes for killing). SNARE proteins are small, membrane-anchored proteins that directly control the membrane fusion events which are necessary for vesicle trafficking. In this project you will investigate how particular SNARE proteins mediate pathogen killing and inflammatory cytokine secretion by macrophages. You will also investigate if SNARE proteins are targeted by virulence factors from important human pathogens (e.g. Salmonella, Legionella, and Mycobacterium) in order to usurp the immune functions of macrophages.
Skill acquisition: a variety of cell biology (e.g. tissue culture, ELISA assays, FACS analysis, and confocal microscopy), biochemical (e.g. affinity purification of proteins and Western blotting), and molecular biology techniques (e.g. Real-Time PCR, cloning, transfection of cells, and siRNA-mediated gene silencing).
Regulation of inflammation and cancer by SNARE proteins
Supervisor: Dr Glen Scholz
Location: Department of Medicine (RMH)
Contact: 8344-3298 Email: glenms@unimelb.edu.au
Following their activation, many cell-surface receptors (e.g. growth factor receptors) are internalized and then trafficked through the endocytic pathway, finally arriving at the lysosome where they are degraded. Such degradation of receptors is thought to be important in limiting their signalling activity. Aberrant trafficking of receptors through the endocytic pathway could therefore potentially result in prolonged signalling and hence altered cellular responses (e.g. prolonged inflammatory responses, cellular hyperproliferation, etc.). SNARE proteins are small, membrane-anchored proteins that directly control the membrane fusion events which are necessary for intracellular vesicle trafficking through the endocytic pathway. In this project you will establish if specific SNARE proteins dictate the biological response of cells to inflammatory stimuli and growth factors by regulating the intracellular trafficking of key signalling receptors.
Skill acquisition: a variety of cell biology (e.g. tissue culture, ELISA assays, FACS analysis, and confocal microscopy), biochemical (e.g. affinity purification of proteins and Western blotting), and molecular biology techniques (e.g. Real-Time PCR, cloning, transfection of cells, and siRNA-mediated gene silencing).
The impact of over-expression and under expression of tissue Plasminogen Activator on epilepsy progression in mice
Supervisors: Dr Nigel Jones, Professor John Hamilton, Professor Terence O’Brien
Location:Department of Medicine (RMH/WH), Clinical Sciences Building, Royal Melbourne Hospital, The University of Melbourne
Contacts: Dr. Nigel Jones T: 8344 6729 E : ncjones@unimelb.edu.au
Professor John Hamilton T: 8344 5480 E: jahami@unimelb.edu.au
Professor Terence O’Brien T: 8344 5490 E: obrientj@unimelb.edu.au
Background.
The processes governing the development of limbic epilepsy are not well understood, but a growing body of literature supports the role of inflammatory mediators in this disease process. One such molecule is tissue Plasminogen Activator (tPA), a clinically used clot-busting enzyme which also has profound effects on cellular physiology in brain regions relevant to temporal lobe epilepsy. These effects, including modulation of cognitive processes, and influencing synaptic connectivity, provide strong rationale to promote tPA as a enzyme which may be involved in development of epilepsy.
Research Plan
The current proposal will investigate the role of tPA signalling in a mouse model of temporal lobe epilepsy. Using genetically engineered mice which are bred to either express an abundance of tPA, or BSC HONOURS PROJECTS 2010 BENCH TO BEDSIDE - MEDICAL RESEARCH 11 a complete lack of tPA, we will determine the direct role of tPA on epilepsy progression. These experiments will incorporate the amygdala kindling model of limbic epilepsy in mice bred in the laboratories of our collaborators. The second aspect of the project will attempt to ascertain the mechanisms by which tPA might influence the progression of disease using immunocytochemical techniques.
Acquired skills will include small animal handling, neurosurgery, amygdala kindling, post-mortem processing, and immunocytochemistry.
Retinal Vascular Calibre and Cardiovascular Disease in Patients with Autoimmune Disease
Supervisor: Dr Sharon Van Doornum
Location: Department of Medicine (RMH/WH)
Contact: Dr Sharon Van Doornum T: 8344 3144 E: svd@unimelb.edu.au
Patients with auto-immune diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) are at increased risk of morbidity and mortality due to cardiovascular disease. It may be that chronic high levels of systemic inflammation initiates and/or accelerates atherosclerosis in patients with autoimmune disease resulting in excess cardiovascular events in these patients. Control of inflammation, along with early detection of cardiovascular disease, are likely to be the keys to reducing the high mortality in patients with autoimmune disease. However, despite this knowledge, predicting persons at risk of cardiovascular disease remains problematic. Thus, there is significant interest in developing new methods that may assist in identifying persons with autoimmune disease who are at higher risk of cardiovascular disease
Two novel and promising methods of early detection of cardiovascular disease are examination of the retinal microcirculation and measurement of arterial stiffness. Application of these techniques in this patient population may be used not only predict cardiovascular disease, but also to gain valuable insights into the role of inflammation in the pathogenesis of vascular disease.
In this study you will investigate the prevalence of retinal vascular abnormalities in a cohort of patients with autoimmune disease, compare this with age and gender matched population controls, and correlate the findings with measures of disease activity, cardiovascular risk factors and arterial stiffness.
The project offers students an opportunity to develop research skills including literature review, effective communication with patients (recruitment, informed consent, clinical assessment), quantitative and qualitative data analysis and epidemiological study skills.