Heparanase induces tissue factor pathway inhibitor expression and extracellular accumulation in endothelial and tumor cells

Summary

Heparanase activity is implicated in cell invasion, tumor metastasis and angiogenesis. Recently, we have reported that heparanase stimulates tissue factor (TF) expression in endothelial and cancer cells, resulting in elevation of coagulation activity. We hypothesized that heparanase regulates other coagulation modulators, and examined the expression and localization of tissue factor pathway inhibitor (TFPI) following heparanase over-expression or exogenous addition. Primary human umbilical vein endothelial cells (HUVEC) and human tumor-derived cell lines were incubated with heparanase, or were stably transfected with heparanase gene-constructs, and TFPI expression and secretion were examined. Heparanase over-expression or exogenous addition stimulated TFPI expression by 2–3 folds. TFPI accumulation in the cell culture medium exceeded in magnitude the observed induction ofTFPI gene transcription reaching 5– to 6-fold increase. Extracellular accumulation of TFPI was evident already 60 min following heparanase addition, prior toTFPI protein induction, and correlated with increased coagulation activity. This effect was found to be independent of heparanase enzymatic activity and interaction with heparan-sulfate, and correlated with reduced TFPI levels on the cell surface. Data were verified in heparanase transgenic mice tissues and plasma. Interaction between heparanase and TFPI was evident by co-immunoprecipitation. Interaction of heparanase with TFPI resulted in its displacement from the surface of the vascular endothelium and in increased pro-coagulant activity. Thus, heparanase facilitates blood coagulation on the cell surface by two independent mechanisms:dissociation ofTFPI from the vascular surface short after local elevation of heparanase levels, and subsequent induction of TF expression.

Keywords

TFPI - heparanase - coagulation - endothelial cells - tumor

• References

• 1 Parish CR, Freeman C, Hulett MD. Heparanase: a key enzyme involved in cell invasion. Biochim Biophys Acta 2001; 1471: M99-108.
• 2 Vlodavsky I, Abboud-Jarrous G, Elkin M. et al. The impact of heparanese and heparin on cancer metastasis and angiogenesis. Pathophysiol Haemost Thromb 2006; 35: 116-127.
• 3 Bar-Sela G, Kaplan-Cohen V, Ilan N. et al. Heparanase expression in nasopharyngeal carcinoma inversely correlates with patient survival. Histopathology 2006; 49: 188-193.
• 4 Koliopanos A, Friess H, Kleeff J. et al. Heparanase expression in primary and metastatic pancreatic cancer. Cancer Res 2001; 61: 4655-4659.
• 5 Rohloff J, Zinke J, Schoppmeyer K. et al. Heparanase expression is a prognostic indicator for postoperative survival in pancreatic adenocarcinoma. Br J Cancer 2002; 86: 1270-1275.
• 6 Gohji K, Hirano H, Okamoto M. et al. Expression of three extracellular matrix degradative enzymes in bladder cancer. Int J Cancer 2001; 95: 295-301.
• 7 Takaoka M, Naomoto Y, Ohkawa T. et al. Heparanase expression correlates with invasion and poor prognosis in gastric cancers. Lab Invest 2003; 83: 613-622.
• 8 Tang W, Nakamura Y, Tsujimoto M. et al. Heparanase: a key enzyme in invasion and metastasis of gastric carcinoma. Mod Pathol 2002; 15: 593-598.
• 9 Shinyo Y, Kodama J, Hongo A. et al. Heparanase expression is an independent prognostic factor in patients with invasive cervical cancer. Ann Oncol 2003; 14: 1505-1510.
• 10 Sato T, Yamaguchi A, Goi T. et al. Heparanase expression in human colorectal cancer and its relationship to tumor angiogenesis, hematogenous metastasis, and prognosis. J Surg Oncol 2004; 87: 174-181.
• 11 Gingis-Velitski S, Zetser A, Flugelman MY. et al. Heparanase Induces Endothelial Cell Migration via Protein Kinase B/Akt Activation. J Biol Chem 2004; 279: 23536-23541.
• 12 Zetser A, Bashenko Y, Edovitsky E. et al. Heparanase induces vascular endothelial growth factor expression: correlation with p38 phosphorylation levels and Src activation. Cancer Res 2006; 66: 1455-1463.
• 13 Elkin M, Ilan N, Ishai-Michaeli R. et al. Heparanase as mediator of angiogenesis: mode of action. Faseb J 2001; 15: 1661-1663.
• 14 Nadir Y, Brenner B, Zetser A. et al. Heparanase induces tissue factor expression in vascular endothelial and cancer cells. J Thromb Haemost 2006; 4: 2443-2451.
• 15 Gingis-Velitski S, Zetser A, Kaplan V. et al. Heparanase Uptake Is Mediated by Cell Membrane Heparan Sulfate Proteoglycans. J Biol Chem 2004; 279: 44084-44092.
• 16 Zetser A, Bashenko Y, Miao H-Q. et al. Heparanase Affects Adhesive and Tumorigenic Potential of Human Glioma Cells. Cancer Res 2003; 63: 7733-7741.
• 17 Zetser A, Levy-Adam F, Kaplan V. et al. Processing and activation of latent heparanase occurs in lysosomes. J Cell Sci 2004; 117: 2249-2258.
• 18 Hulett MD, Hornby JR, Ohms SJ. et al. Identification of active-site residues of the pro-metastatic endoglycosidase heparanase. Biochemistry 2000; 39: 15659-15667.
• 19 Levy-Adam F, Abboud-Jarrous G, Guerrini M. et al. Identification and characterization of heparin/heparan sulfate binding domains of the endoglycosidase heparanase. J Biol Chem 2005; 280: 20457-20466.
• 20 Nardella C, Lahm A, Pallaoro M. et al. Mechanism of activation of human heparanase investigated by protein engineering. Biochemistry 2004; 43: 1862-1873.
• 21 Zcharia E, Metzger S, Chajek-Shaul L T. et al. Transgenic expression of mammalian heparanase uncovers physiological functions of heparan sulfate in tis- sue morphogenesis, vascularization, and feeding behavior. FASEB J 2004; 18: 252-263.
• 22 Ho G, Broze Jr GJ, Schwartz AL. Role of heparan sulfate proteoglycans in the uptake and degradation of tissue factor pathway inhibitor-coagulation factor Xa complexes. J Biol Chem. 1997: 16838-16844.
• 23 Fuki II, Iozzo RV, Williams KJ. Perlecan heparan sulfate proteoglycan. A novel receptor that mediates a distinct pathway for ligand catabolism. J Biol Chem 2000; 275: 25742-50.
• 24 Lwaleed BA, Bass PS. Tissue factor pathway inhibitor: structure, biology and involvement in disease. J Pathol 2006; 208: 327-339.
• 25 Iversen N, Lindahl AK, Abildgaard U. Elevated TFPI in malignant disease: relation to cancer type and hypercoagulation. Br J Haematol 1998; 102: 889-895.
• 26 Gerlach R, Scheuer T, Bohm M. et al. Increased levels of plasma tissue factor pathway inhibitor in patients with glioblastoma and intracerebral metastases. Neurol Res 2003; 25: 335-338.
• 27 Shafat I, Zcharia E, Nisman B. et al. An ELISA method for the detection and quantification of human heparanase. Biochem Biophys Res Commun 2006; 341: 958-963.
• 28 Vlodavsky I, Eldor A, Haimovitz-Friedman A. et al. Expression of heparanase by platelets and circulating cells of the immune system: possible involvement in diapedesis and extravasation. Invasion Metastasis 1992; 12: 112-127.
• 29 Haim N, Lanir N, Hoffman R. et al. Acquired activated protein C resistance is common in cancer patients and is associated with venous thromboembolism. Am J Med 2001; 110: 91-96.
• 30 Hoke M, Kyrle PA, Minar E. et al. Tissue factor pathway inhibitor and the risk of recurrent venous thromboembolism. Thromb Haemost 2005; 94: 787-790.
• 31 Sarig G, Blumenfeld Z, Leiba R. et al. Modulation of systemic hemostatic parameters by enoxaparin during gestation in women with thrombophilia and pregnancy loss. Thromb Haemost 2005; 94: 980-985.
• 32 Mousa SA, Linhardt R, Francis JL. et al. A. Antimetastatic effect of a non-anticoagulant low-molecular- weight heparin versus the standard low-molecularweight heparin, enoxaparin. Thromb Haemost 2006; 96: 816-21.
• 33 Hostettler N, Naggi A, Torri G. et al. P-selectin- and heparanase-dependent antimetastatic activity of nonanticoagulant heparins. FASEB J 2007; 21: 3562-3572.
• 34 Liu T, Scallan CD, Broze Jr GJ. et al. Improved coagulation in bleeding disorders by Non-Anticoagulant Sulfated Polysaccharides (NASP). Thromb Haemost 2006; 95: 68-76.
• 35 Naggi A, Casu B, Perez M. et al. Modulation of the heparanase-inhibiting activity of heparin through selective desulfation, graded N-acetylation, and glycol splitting. J Biol Chem 2005; 280: 12103-12113.