Group Leader: Chris Parish

Overview:

The major research interest of the Cancer and Vascular Biology Group is the molecular basis of cell adhesion, cell migration and cell invasion, with a particular emphasis on the immune system, tumour metastasis and the growth of new blood vessels (angiogenesis). Of particular focus is the role of anionic carbohydrates, such as heparan sulfate (HS), in these processes. The Group is led by Professor Chris Parish, and comprises 3 laboratories, the Cellular Laboratory headed by Chris, the Molecular Mechanisms Laboratory headed by Dr Mark Hulett, and the Matrix Biology Laboratory headed by Dr Craig Freeman.

Model structure of the human heparanase
active site domain

Research Projects:

Role of heparanase in cell invasion and angiogenesis
The major barrier for invading tumour cells, migrating leukocytes, and growing blood vessels (endothelial cells) is the basement membrane (BM) that surrounds the vessels, and the extracellular matrix (ECM) which forms a scaffold in tissues to hold cells together. The BM and ECM are composed of an interlocking network of proteins and complex carbohydrates, and for cells to breach this barrier, they deploy a battery of enzymes that break down these proteins and carbohydrate components. The major carbohydrate is heparan sulphate (HS), which acts as the glue to maintain the integrity of the BM and ECM. The enzyme responsible for cleaving HS, heparanase, has been shown to play a key role in the degradation of the BM and ECM, and its activity strongly correlates with the metastatic capacity of tumour cells and the migratory capacity of leukocytes and endothelial cells. HS in the ECM also binds a number of angiogenic growth factors, and the release of these by heparanase promotes angiogenesis and tumour growth. Following our recent cloning of mammalian heparanase, we have been able to develop the tools to investigate how heparanase functions at the molecular level and to directly determine the role of heparanase in cell invasion, angiogenesis and inflammation.

Role of histidine-rich glycoprotein (HRG) in regulating immune complex clearance and cell invasion
The group has also been studying the plasma protein, histidine-rich glycoprotein (HRG), particularly examining the ability of the protein to inhibit cell adhesion by masking cell surface carbohydrates. Recently, the group demonstrated that HRG plays an important role in the immune system by interacting with complement components and by preventing the insolubilisation of complexes between antibody and antigen (termed immune complexes). In fact HRG also assists in the uptake of these complexes by phagocytic cells. Thus HRG is probably a key molecule in regulating complement activity and in aiding the elimination of immune complexes from the circulation. In fact, deficiencies in HRG may lead to immune complex-associated diseases such as rheumatoid arthritis (RA) and systemic lupus erythematosis (SLE). In a related study we have shown that HRG can tether plasmin/plasminogen to the surface of cells and potentially aid cell invasion. Thus HRG represents a multifunctional protein that appears to play an important role in the immune system, inflammation and wound healing. A major focus of the group in the future is to better understand the functional significance of this intriguing plasma protein.

Development of a novel tumour vaccine
In collaboration with Dr Paul Foster's group in the Division of Molecular Biosciences, JCSMR, a new approach to cancer immunotherapy has been developed. Currently most attempts at cancer immunotherapy involve the generation of CD8+ cytotoxic T lymphocytes (CTLs) against tumour-specific antigens. Recently we demonstrated that tumour-specific CD4+ T cells, that exhibit a cytokine secretion profile characteristic of Th2 cells, are capable of clearing established lung and visceral metastases of a B16 melanoma that is resistant to CTL lysis. Clearance of the lung metastases by Th2 cells was found to be dependent on degranulating eosinophils, with the eosinophil chemokine, eotaxin, playing an essential role. In contrast, tumour-specific CD4+ Th1 cells, that recruited macrophages into the tumour, had no effect on tumour growth. This work provides the basis for a new approach to cancer vaccination that is effective against CTL-resistant tumours and is, potentially, less susceptible to immune evasion.

Th2 inhibition of tumour metastasis. The effect of adoptive transfer of ovalbumin-specific Th2 cells on the growth of established lung metastasis from a highly metastatic mouse B16-F1 melanoma secreting ovalbumin in a mouse model is shown.

Group Members:

Key Publications:

Altin J G, van Broekhoven C L and Parish C R: (2004) Targeting dendritic cells with antigen-containing liposomes: antitumour immunity. Expert Opinion on Biological Therapy 4, 1735-1747.

Jones A L, Hulett M D, Altin J G, Hogg P and Parish C R: (2004) Plasminogen is tethered with high affinity to the cell surface by the plasma protein, histidine-rich glycoprotein. The Journal of Biological Chemistry 279, 38267-38276.

Jones A L, Hulett M D and Parish C R: (2004) Histidine-rich glycoprotein binds to cell-surface heparan sulfate via its N-terminal domain following Zn2+ chelation. The Journal of Biological Chemistry 279, 30114-30122.

Khachigian L M and Parish C R: (2004) Phosphomannopentaose sulfate (PI-88): Heparan sulfate mimetic with clinical potential in multiple vascular pathologies. Cardiovascular Drug Reviews 22, 1-6.

Levidiotis V, Freeman C, Punler M, Martinello P, Creese B, Ferro V, van der Vlag J, Berden J H, Parish C R and Power D A: (2004) A synthetic heparanase inhibitor reduces proteinuria in passive Heymann nephritis. Journal of the American Society of Nephrology 15, 2882-2892.

Levidiotis V, Freeman C, Tikellis C, Cooper M E and Power D A: (2004) Heparanase is involved in the pathogenesis of proteinuria as a result of glomerulonephritis. Journal of the American Society of Nephrology 15, 68-78.

Nyberg K, Ekblad M, Bergström T, Freeman C, Parish C R, Ferro V and Tryala E: (2004) The low molecular weight heparan sulfate-mimetic, PI-88, inhibits cell-to-cell spread of herpes simplex. Antiviral Research 63, 15-24.

van Broekhoven C L, Parish C R, Demangel C, Britton W J and Altin J G: (2004) Targeting dendritic cells with antigen-containing liposomes: A highly effective procedure for induction of antitumor immunity and for tumor immunotherapy. Cancer Research 64, 4357-4365.

Xie W, McCahon P, Jakobsen K and Parish C: (2004) Evaluation of the ability of digital infrared imaging to detect vascular changes in experimental animal tumours. International Journal of Cancer 108, 790-794.

Francis, D.J., Parish, C.R., McGarry,Y. M., Santiago, F.S., Brown, K.J., Bingley, J.A., Hayward, I.P., Cowden, W.B., Campbell, J.H., Campbell, G.R., Chesterman, C.N. and Khachigian, L.M. (2003)
Blockade of vascular smooth muscle cell proliferation and intimal thickening after balloon injury by the sulfated oligosaccharide PI-88: phosphomannopentaose sulfate directly binds FGF-2, blocks cellular signaling and inhibits proliferation. Circ Res 92, E70-E77.

Mattes, J., Hulett, M., Xie, W., Hogan, S., Rothenberg, M.E., Foster, P. and Parish, C.R. (2003)
Immunotherapy of cytotoxic T cell resistant tumors by T helper 2 cells: an eotaxin-1 and STAT-6-dependent process. J.Exp.Med. 197, 387-393.

Parish, C.R. (2003)
Cancer immunotherapy: The past, the present and the future. Immunol. Cell Biol. - 81, 106-113.

Armitt, D.J., Banwell, M.G., Freeman, C. and Parish, C.R. (2002)
C-glycoside formation via Lewis-acid promoted reaction of O-glycosylimidates with pyrroles. J. Chem. Soc., Perkin Trans. 1, 1743-1745.

Manderson AP, Pickering MC, Botto M, Walport MJ and Parish CR (2001)
Continual low-level activation of the classical complement pathway. J. Exp. Med. 194, 745-756.

Hindmarsh, E.J., Staykova, M.A, Willenborg, D.O. and Parish, C.R. (2001)
Cell surface expression of the 300 kDa mannose-6-phosphate receptor by activated T lymphocytes. Immunol. Cell Biol, 79, 436-443.

Wall D, Douglas S, Ferro V, Cowden W and Parish C (2001)
Characterisation of the anticoagulant properties of a range of structurally diverse sulfated oligosaccharides. Thromb. Res. 103, 325-335.

Parish CR, Freeman C and Hulett MD (2001)
Heparanase: a key enzyme involved in cell invasion. Biochem. Biophys. Acta 1471, M99-M108.

van Broekhoven, C.L., Parish, C.R., Vassiliou, G. and Altin, J. (2000).
Engrafting costimulator molecules onto tumor cell surfaces with chelator lipids: a potentially convenient approach in cancer vaccine development. J. Immunol. 164, 2433-2443.

Hulett MD, Hornby JR, Ohms J, Zeugg J, Freeman C, Gready JE and Parish CR (2000)
Identification of active site residues of the pro-metastatic endoglycosidase heparanase. Biochemistry 39, 15659-15667.

Freeman, C., Browne, A.M. and Parish, C.R. (1999)
Evidence that platelet and tumour heparanases are similar enzymes. Biochem. J., 342, 361-368.

Hulett, M.D., Freeman, C., Hamdorf, B.J., Baker, R.T., Harris, M.J. and Parish, C.R. (1999).
Cloning of mammalian heparanase, an important enzyme in tumour invasion and metastasis. Nature Med. 5, 803-809.

Parish, C.R., Freeman, C., Brown, K.J., Francis, D. and Cowden, W.B. (1999).
Identification of sulfated oligosaccharide-based inhibitors of tumor growth and metastasis using novel in vitro assays for angiogenesis and heparanase activity. Cancer Res. 59, 3433-3441.

Gorgani, N.N., Parish, C.R., and Altin, J.G. (1999).
Differential binding of histidine-rich glycoprotein (HRG) to human IgG subclasses and IgG molecules containing kappa and lambda light chains. J. Biol. Chem., 274, 29633-29640.

Gorgani, N.N., Altin, J.G. and Parish, C.R. (1999).
Histidine-rich glycoprotein regulates the binding of IgG and immune complexes to monocytes. Int. Immunol., 11, 1275-1282.

Grants 2004
National Health and Medical Research Council Program Grant
Professor C Chesterman (UNSW), Dr R Andrews (Monash), Dr M Berndt (Monash),
Professor B Chong (UNSW), Professor P Hogg (UNSW), Dr M Hulett, A/Professor L Khachigian (UNSW) and Professor C Parish
Vascular Biology
$2,000,000

NSW Cancer Council
Professor C Parish, Dr J Altin (BaMBi), Dr P Hogg (UNSW) and Professor C Chesterman (UNSW)
Tumour angiogenesis
$196,000
Australian Research Council Centre of Excellence Grant
Professor P Gresshoff, Professor C Beveridge, Dr B Carroll, Professor B Rolfe, Professor C.Parish, Dr M Djordjevic, Dr G Weiller, Dr U Mathesius, Dr R Rose, Professor M Singh and
Dr P Bhalla
Integrated Legume Research
$2,000,000

Australian Research Council - LIEF Grant
Dr B Garner, Professor C Parish, Dr C Freeman, Dr PP Gray, Dr MJ Davies and Dr M. Guilhaus
A glycobiology facility for glycoconjugate analysis and oligosaccharide sequencing
$343,608

Viertel Senior Medical Research Fellowship
Dr M Hulett
Regulators of cell invasion and angiogenesis
$165,000 (per year 2002-2006)

National Health and Medical Research Council Equipment Grant
Dr M Hulett
Amaxa Nucleofector Device
$32,000

Funding Sources:

NH&MRC of Australia, Australian Research Council, NSW Cancer Council, DETYA, Sylvia and Charles Viertel Charitable Foundation

The Cancer and Vascular Biology Group has had considerable experience in designing sulfated oligosaccharide-based compounds as drug candidates, this research being supported by a commercial partner, Progen Industries, Brisbane. Sulfated oligosaccharide-based inhibitors of the heparanase enzyme have been synthesised and identified, with a sulfated oligosaccharide, termed PI-88, being found to be a potent inhibitor of angiogenesis and heparanase activity.

Preclinical testing has shown that PI-88 can inhibit primary tumour growth and metastasis of a number of cancer types. By the end of 2000 the drug had successfully completed phase I clinical trials in healthy volunteers and cancer patients, with phase II trials in cancer patients commencing in 2001.

More recently, a very productive collaboration has been initiated with Professor Martin Banwell, Research School of Chemistry, ANU in which sulfated pseudo-sugars are being synthesized as heparan sulfate mimetics. This project is also being funded by Progen Industries. Although the initial aim of this collaboration is to produce better heparanase inhibitors, a number of sulfated pseudo-sugars have been identified that selectively inhibit certain protein-heparan sulfate interactions. Such drugs have potential as anti-angiogenic, anticoagulant, antiviral and antilipaemic agents.

Patents:

Inventors & Stage: Cowden WB, Willenborg DO, Parish CR. (PCT/AU88/0017)
Description: Compounds having anti-metastatic and/or anti-inflammatory activity

Inventors & Stage: Cowden WB, Willenborg DO, Parish CR. (PCT/AU89/00183)
Description: Use of castanospermine as an anti-inflammatory agent

Inventors & Stage: Parish CR, Cowden WB, Willenborg DO. (PCT/AU89/00350)
Description: Phosphosuger-based anti-inflammatory drugs

Inventors & Stage: Parish CR. (PCT/AU93/00558)
Description: Angiogenesis inhibitory antibodies

Inventors & Stage: Parish CR, Brown KJI, Maynes SF, Bezos A. (PCT/AU95/00105)
Description: In vitro angiogenesis assay

Inventors & Stage: Parish CR, Cowden WB. (PCT/AU96/00238)
Description: Preparation and use of sulfated oligosaccharides

Inventors & Stage: Freeman CG, Parish CR. (PCT/AU97/00452)
Description: Detection of mammalian heparanase activity and purification of mammalain heparanase

Inventors & Stage: Cowden WB, Willenborg DO, Parish CR. (PCT/AU97/00250)
Description: Phosphosugars and phosphosugar containing compounds having anti-inflammatory activity.

Inventors & Stage: Parish CR, Cowden WB. (PCT/AU98/00151)
Description: Sulfated oligosaccharides having anticoagulant/antithrombotic activity.

Inventors & Stage: Altin G, Burns CJ, Pace RJ, Parish CR, Fiddes RJ. (PCT/AU98/00417)
Description: Receptor/ligand biosensor

Inventors & Stage: Cowden WB, Francis DJ, Parish CR. (PCT/AU98/00707)
Description: Use of sulfated oligosaccharides as inhibitors of cardiovascular disease.

Inventors & Stage: Cowden WB, Parish CR. (PCT/AU98/00844)
Description: Use of sulfated oligosaccharides in lowering blood triglyceride levels.

Inventors & Stage: Freeman CG, Hulett MD, Parish CR, Hamdorf BJ. (PCT/AU98/00898)
Description: Isolated nucleic acid molecule encoding mammalian endoglucuronidase and uses therefor.

Inventors & Stage: Altin JG, Parish CR. (PCT/AU00/00397)
Description: Model membrane systems.

Inventors & Stage: Parish CR. and Cabalda-Crane V. (PC/AU01/00877)
Description: Method of identifying cancer markers and uses therefor in the diagnosis of cancer.

Inventors & Stage: Banwell MG, Edwards AJ, Ferro V, Freeman C, Liu L and Parish CR. (PCT number pending) Lodged 3 July 2001.
Description: Linked cyclitols and their polysulfated derivatives.

Inventors & Stage: Parish CR and Cabalda-Crane V (US Provisional Patent Application) Lodged 3 August, 2001.
Description: A novel cancer marker and uses therefore in the diagnosis of cancer.

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