JAK-2 V617F mutation increases heparanase procoagulant activity

Inna Kogan; Dafna Chap; Ron Hoffman; Elena Axelman; Benjamin Brenner; Yona Nadir

doi:10.1160/TH15-04-0320

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2016; 115(01): 73-80
DOI: 10.1160/TH15-04-0320

Coagulation and Fibrinolysis

Schattauer GmbH Schattauer

JAK-2 V617F mutation increases heparanase procoagulant activity

Inna Kogan

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

,

Dafna Chap

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

,

Ron Hoffman

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

,

Elena Axelman

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

,

Benjamin Brenner

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

,

Yona Nadir

¹Thrombosis and Hemostasis Unit, Rambam Health Care Campus, The Bruce Rappaport Faculty of Medicine, Technion Haifa, Israel

› Author Affiliations

Abstract

Summary

Patients with polycythaemia vera (PV), essential thrombocythaemia (ET) and primary myelofibrosis (PMF) are at increased risk of arterial and venous thrombosis. In patients with ET a positive correlation was observed between JAK-2 V617F mutation, that facilitates erythropoietin receptor signalling, and thrombotic events, although the mechanism involved is not clear. We previously demonstrated that heparanase protein forms a complex and enhances the activity of the blood coagulation initiator tissue factor (TF) which leads to increased factor Xa production and subsequent activation of the coagulation system. The present study was aimed to evaluate heparanase procoagulant activity in myeloproliferative neoplasms. Forty bone marrow biopsies of patients with ET, PV, PMF and chronic myelogenous leukaemia (CML) were immunostained to heparanase, TF and TF pathway inhibitor (TFPI). Erythropoietin receptor positive cell lines U87 human glioma and MCF-7 human breast carcinoma were studied. Heparanase and TFPI staining were more prominent in ET, PV and PMF compared to CML. The strongest staining was in JAK-2 positive ET biopsies. Heparanase level and procoagulant activity were higher in U87 cells transfected to over express JAK-2 V617F mutation compared to control and the effect was reversed using JAK-2 inhibitors (Ruxolitinib, VZ3) and hydroxyurea, although the latter drug did not inhibit JAK-2 phosphorylation. Erythropoietin increased while JAK-2 inhibitors decreased the heparanase level and procoagulant activity in U87 and MCF-7 parental cells. In conclusion, JAK-2 is involved in heparanase up-regulation via the erythropoietin receptor. The present findings may potentially point to a new mechanism of thrombosis in JAK-2 positive ET patients.

Keywords

Heparanase - myeloproliferativ - JAK-2 - coagulation

Full Text

References

References
1 Kreher S, Ochsenreither S, Trappe RU. et al. Prophylaxis and management of venous thromboembolism in patients with myeloproliferative neoplasms: consensus statement of the Haemostasis Working Party of the German Society of Hematology and Oncology (DGHO), the Austrian Society of Hematology and Oncology (OGHO) and Society of Thrombosis and Haemostasis Research (GTH e. V.). Ann Hematol 2014; 93: 1953-1963.
2 Barbui T, Finazzi G, Falanga A. Myeloproliferative neoplasms and thrombosis. Blood 2013; 122: 2176-2184.
3 Lussana F, Caberlon S, Pagani C. et al. Association of V617F Jak2 mutation with the risk of thrombosis among patients with essential thrombocythaemia or idiopathic myelofibrosis: a systematic review. Thromb Res 2009; 124: 409-417.
4 Freeman C, Parish CR. Human platelet heparanase: purification, characterization and catalytic activity. Biochem J 1998; 330: 1341-1350.
5 Pikas DS, Li JP, Vlodavsky I. et al. Substrate specificity of heparanases from human hepatoma and platelets. J Biol Chem 1998; 273: 18770-18777.
6 Parish CR, Freeman C, Hulett MD. Heparanase: a key enzyme involved in cell invasion. Biochim Biophys Acta 2001; 1471: M99-108.
7 Vlodavsky I, Friedmann Y. Molecular properties and involvement of heparanase in cancer metastasis and angiogenesis. J Clin Invest 2001; 108: 341-347.
8 Nadir Y, Brenner B, Zetser A. et al. Heparanase induces tissue factor expression in vascular endothelial and cancer cells. J Thromb Haemost 2006; 04: 2443-2451.
9 Nadir Y, Brenner B, Gingis-Velitski S. et al. Heparanase induces tissue factor pathway inhibitor expression and extracellular accumulation in endothelial and tumour cells. Thromb Haemost 2008; 99: 133-141.
10 Nadir Y, Brenner B, Fux L. et al. Heparanase enhances the generation of activated factor X in the presence of tissue factor and activated factor VII. Haematologica 2010; 95: 1927-1934.
11 Nadir Y, Kenig Y, Drugan A. et al. An assay to evaluate heparanase procoagulant activity. Thromb Res 2011; 128: e3-8.
12 Peled E, Rovitsky A, Axelman E. et al. Increased heparanase level and procoagulant activity in orthopedic surgery patients receiving prophylactic dose of enoxaparin. Thromb Res 2012; 130: 129-134.
13 Matan M, Axelman E, Brenner B. et al. Heparanase procoagulant activity is elevated in women using oral contraceptives. Hum Reprod 2013; 28: 2372-2380.
14 Nadir Y, Sarig G, Axelman E. et al. Heparanase procoagulant activity is elevated and predicts survival in non-small cell lung cancer patients. Thromb Res 2014; 134: 639-642.
15 Nadir Y, Saharov G, Hoffman R. et al. Heparanase procoagulant activity, factor Xa, and plasminogen activator inhibitor 1 are increased in shift work female nurses. Ann Hematol 2015; 94: 1213-1219.
16 Elkin M, Cohen I, Zcharia E. et al. Regulation of heparanase gene expression by estrogen in breast cancer. Cancer Res 2003; 63: 8821-8826.
17 Han J, Woytowich AE, Mandal AK. et al. Heparanase upregulation in high glucose-treated endothelial cells is prevented by insulin and heparin. Exp Biol Med 2007; 232: 927-934.
18 Vardiman JW, Thiele J, Arber DA. et al. The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes. Blood 2009; 114: 937-951.
19 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.
20 Zetser A, Levy-Adam F, Kaplan V. et al. Processing and activation of latent heparanase occurs in lysosomes. J Cell Sci 2004, 2004; 117: 2249-2258.
21 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.
22 Zetser A, Bashenko Y, Miao H-Q. et al. Heparanase Affects Adhesive and Tumourigenic Potential of Human Glioma Cells. Cancer Res 2003; 63: 7733-7741.
23 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.
24 Nadir Y, Kenig Y, Drugan A. et al. Involvement of heparanase in vaginal and cesarean section deliveries. Thromb Res 2010; 126: e444-450.
25 Nadir Y, Henig I, Naroditzky I. et al. Involvement of Heparanase in early pregnancy losses. Thromb Res 2010; 125: 251-257.
26 Glaspy J. Current status of use of erythropoietic agents in cancer patients. Semin Thromb Hemost 2014; 40: 306-312.
27 Ward AC, Touw I, Yoshimura A. The Jak-Stat pathway in normal and perturbed hematopoiesis. Blood 2000; 95: 19-29.
28 Parganas E, Wang D, Stravopodis D. et al. Jak2 is essential for signalling through a variety of cytokine receptors. Cell 1998; 93: 385-395.
29 Silvennoinen O, Witthuhn BA, Quelle FW. et al. Structure of the murine Jak2 protein-tyrosine kinase and its role in interleukin 3 signal transduction. Proc Natl Acad Sci USA 1993; 90: 8429-8433.
30 Tolcher AW, Giusti RM, O’Shaughnessy JA. et al. Arterial thrombosis associated with granulocyte-macrophage colony-stimulating factor (GM-CSF) administration in breast cancer patients treated with dose-intensive chemotherapy: a report of two cases. Cancer Invest 1995; 13: 188-192.
31 O’Reilly SE, Gelmon KA, Onetto N. et al. Phase I trial of recombinant human granulocyte-macrophage colony-stimulating factor derived from yeast in patients with breast cancer receiving cyclophosphamide, doxorubicin, and fluorouracil. J Clin Oncol 1993; 11: 2411-2416.
32 Usuki K, Iki S, Endo M. et al. Influence of thrombopoietin on platelet activation in myeloproliferative disorders. Br J Haematol 1997; 97: 530-537.
33 Cuker A. Toxicities of the thrombopoietic growth factors. Semin Hematol 2010; 47: 289-298.
34 Cortelazzo S, Finazzi G, Ruggeri M. et al. Hydroxyurea for patients with essential thrombocythemia and a high risk of thrombosis. N Engl J Med 1995; 332: 1132-1136.
35 Maugeri N, Giordano G, Petrilli MP. et al. Inhibition of tissue factor expression by hydroxyurea in polymorphonuclear leukocytes from patients with myeloproliferative disorders: a new effect for an old drug?. J Thromb Haemost 2006; 04: 2593-2598.