Molecular Immunology and Cancer

Group Leader: Dr Mark Hulett

---

Overview of Research

Cancer Metastasis, Angiogenesis, and Inflammation

The ability of malignant tumour cells to escape from primary tumour sites and spread through the circulation to other sites in the body (metastasis) is what makes cancer such a deadly disease (Figure 1). The essential processes in metastasis are cell invasion - where tumour cells move into and out of the vasculature, and angiogenesis - where new blood/lymph vessels are formed in and around the tumour that provide an escape route and also supply nutrients for tumour growth. Cell invasion is also a critical event in the migration of white blood cells of the immune system (leukocytes) to sites of inflammation to combat infections. Understanding the molecular basis of cell invasion and angiogenesis is vital to develop strategies to combat cancer spread and inflammatory disease.



The major barrier for invading tumour cells, migrating leukocytes, and growing blood vessels (endothelial cells) is the basement membrane (BM), which 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. A 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 roll 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.

In contrast to many of the proteases involved in degrading the protein component of the BM and ECM, until recently knowledge of the exact structure of heparanase remained elusive. In collaboration with Prof Chris Parish at the John Curtin School of Medical Research, we were the first to clone the first mammalian (human, mouse, rat) heparanase genes (Hulett et al., 1999. Nature Medicine 5:803-809. ). This opened the door to develop the tools to enable the direct study of the role of the enzyme in cell invasion and angiogenesis, which is the main focus of our research.




We have since shown that (i) the cloned heparanase enzyme is the dominant heparanase in mammalian tissues, making it an extremely attractive drug target, (ii) the enzyme is synthesised as an inactive pro form that requires proteolytic processing for activity, and (iii) identified the active site of the enzyme and proposed a model structure of the enzyme (Figure 2), (iv) defined key transcription factors that regulate heparanase gene expression in disease, and (v) generated heparanase conditional knockout mice. We are currently working towards (i) further understanding the molecular basis of heparanase function at the structural level, (ii) defining the dysregulation of heparanase gene expression in cancer and inflammatory disease, and (iii) using heparanase conditional knockout mice to define the precise role and contribution of heparanase in tumour progression, inflammation, and vascular injury. 
Our overall goal is to better understand both the biology and structure of heparanase to enable the development of specific inhibitors of the enzyme, which will hopefully lead to new drugs for the treatment of tumour metastasis, angiogenesis, and inflammatory diseases.

Group Members

Key Publications

Kavallaris M, Meachem SJ, Hulett MD, West CM, Pitt RE, Chesters JJ, Laffan WS, Boreham PR, Khachigian LM. 2008. Perceptions in health and medical research careers: the Australian Society for Medical Research Workforce Survey. Med J Aust. 2008 May 5;188(9):520-524.

Quah, B., Barlow, V.P., McPhun, V., Matthaei, K.I., HULETT, M.D. and Parish, C.R. 2008. Bystander B Cells Rapidly Acquire Antigen Receptors from Activated B Cells by Membrane Transfer: a Novel Mechanism for Enhancing Specific Antigen Presentation. Proc. Natl. Acad. Sci. ( USA) 105: 4259-264

Wood, R. and HULETT M.D. 2007. Cell surface expressed cation-independent mannose-6 phosphate receptor (CD222) binds enzymatically active heparanase independently of mannose-6 phosphate to promote extracellular matrix degradation. J. Biol. Chem. 283:4165-176.

de Mestre, A.M., Staykova, M., Hornby, J., Willenborg, D. and HULETT M.D. 2007. The expression of the heparan sulphate degrading enzyme heparanase is induced in infiltrating CD4+ T cells in experimental autoimmune encephalomyelitis and regulated at the level of transcription by early growth response gene 1 . J. Leuk. Biol. 82:1289-300.

de Mestre, A.M., Soe-Htwe, T., Sutcliffe, E.L., Rao, S., Pagler, E.B., Hornby, J.R. and HULETT, M.D. 2007. Regulation of mouse Heparanase gene expression in T lymphocytes and tumour cells. Immunol. Cell Biol. 85:205-214.

Forbes, E., HULETT, M.D., Ahens, R., Smart, V., Matthaei, K.I., Brandt, E., Rothenberg, M.E., Tang, M., Foster, P.S. and Hogan, S.P. 2006. ICAM-1 dependent pathways regulate colonic eosinophilic inflammation. J. Leuk. Biol. 80:330-41.

Abby, J.L., HULETT M.D. and O’Neill H. 2006. Cell surface expression of a peptide encoded by the un-rearranged TCR-Vbeta 8.2 gene. Mol. Immunol. 43:1408-1417.

Jones, A.L., Poon, I., HULETT, M.D. and Parish, C. R. 2005. Histidine-rich glycoprotein specifically binds to necrotic cells, via its amino-terminal domain, and facilitates necrotic cell phagocytosis. J. Biol. Chem. 280:35733-35741.

Jones, A.L., HULETT, M.D. and Parish, C. R. 2005. Histidine-rich glycoprotein: A novel adaptor protein in plasma that modulates the immune, vascular and coagulation systems. Immunol Cell Biol. 83:106-18.

de Mestre, A.M, Rao, S., Hornby, J.R., Soe Htwe, Khachigian, L.M. and HULETT M.D. 2005. EGR1 regulates heparanase gene transcription in tumour cells. J. Biol. Chem. 280:35136-35147.

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. J Biol. Chem. 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 amino-terminal domain following Zn2+ chelation. J. Biol. Chem. 279:30114-30122.

de Mestre, A.M., Khachigian L.M., Santiago, F.S., Staykova, M.A. and HULETT M.D. 2003. Regulation of inducible heparanase gene transcription in activated T cells by early growth response 1. J. Biol. Chem. 278:50377-50385.

Mattes, J., HULETT, M.D., 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 and STAT6 dependent process. J. Exp. Med. 197: 387-393.

Mackay, G.A., HULETT, M.D., Cook, J.P.D., Trist, H.M., Henry, A.J., Beavil, A.J., Beavil, R.L., Sutton, B.J., Hogarth, P.M. and Gould H.J. 2002. Mutagenesis within human FceRIa differentially affects human and murine IgE. J. Immunol., 168, 1787-1795.

Parish, C. R., C. Freeman and M.D. HULETT. 2001. Heparanase: a key enzyme involved in cell invasion. Biochimica et Biophysica Acta - Reviews in Cancer 1471: M99-M108.

HULETT, M.D., Pagler, E. and Hornby J.R. 2001. Cloning and characterisation of a mouse homologue of the human hematopoietic cell-specific four-transmembrane gene HTm4. Immunol Cell Biol., 79, 345-349.

HULETT, M.D., E. Pagler, J. Hornby, P.M. Hogarth, H.J. Eyre, E. Baker, J. Crawford, G.R. Sutherland and Parish, C.R. 2001. Isolation, tissue distribution, and chromosomal localisation of a novel testis-specific human four-transmembrane gene related to CD20 and FceRIb. Biochem. Biophys. Res. Commun., 280, 374-379.

HULETT, M.D., J. Hornby, S. Ohms, J. Zeugg, C. Freeman, J. Greedy and C.R. Parish. 2000. Identification of active site residues of the pro-metastatic endoglycosidase heparanase. Biochemistry, 39, 15659-15667.

Rigby, L.J., H. Trist, H., V.C. Epa, J. Snider, M.D. HULETT and P.M. Hogarth. 2000. Monoclonal antibodies and synthetic peptides define the active site of FceRI and a potential receptor antagonist. Allergy, 55, 609-619.

Rigby, L.J., V.C. Epa, G.A. Mackay, M.D. HULETT, B.J. Sutton, H.J. Gould and P.M. Hogarth. 2000. Domain one of the high affinity Fc epsilon receptor, FceRI, regulates binding to IgE through its interface with domain two. J. Biol. Chem., 275, 9664-9672.

HULETT, M.D. and C.R. Parish. 2000. Murine Histidine-rich Glycoprotein: Cloning, characterisation and cellular origin. Immunol. Cell Biol., 78, 280-287.

Baker, E., J. Crawford, G.R. Sutherland, C. Freeman, C.R. Parish, and M.D. HULETT. 1999. Human Hpa. Endogylcosidase heparanase. Map postion 4q21.3. Chromosome Research 7, 319.

HULETT, M.D., C. Freeman, B.J. Hamdorf, R.T. Baker, M.J. Harris, and C.R. Parish. 1999. Cloning of mammalian heparanase: A key enzyme in tumour invasion and metastasis. Nature Medicine 5:803-809.

Maxwell, K.F., M.S. Powell, M.D. HULETT, P.A. Barton, I.F.C. McKenzie, T.P. Garrett and P.M. Hogarth. 1999. Crystal structure of the human leukocyte Fc receptor FcgRIIa. Nature Structural Biology 6:437-442.

HULETT, M.D. E. Witort, R. Brinkworth, I.F.C. McKenzie and P.M. Hogarth. 1999. Fine structure analysis of the interaction of FceRI with IgE. J. Biol. Chem. 274:13345-13352.

Wines, B. D., M.D. HULETT, G.P. Jamieson, H.M. Trist, J.M. Spratt, and P.M. Hogarth. 1999. Identification of residues in the first domain of Human FcaR essential for interaction with IgA. J. Immunol. 162:2146-2153.