Latent Heparanase Facilitates VLA-4–Mediated Melanoma Cell Binding and Emerges As a Relevant Target of Heparin in the Interference with Metastatic Progression

Abstract

Heparanase is an endo-β-glucuronidase that enzymatically cleaves heparan sulfates (HS) and heparan sulfate proteoglycan (HSPG) structures. Heparanase expression levels by tumors were correlated with cell invasion, angiogenic activity, and poor prognosis. Heparanase can also possess pro-tumorigenic effects independent of its enzymatic activity. Using human melanoma MV3 cells, we demonstrate that latent heparanase activates in a tightly temporary-regulated manner the binding function of the integrin very late antigen-4 (VLA-4), an important component in the metastatic spread of melanoma cells. shRNA-mediated knockdown of syndecan-4 (SDC-4) indicated that this proteoglycan is the key element to convey heparanase binding via focal adhesion complex formation, detected by vinculin staining, to an upregulated VLA-4 binding function. This inside-out signaling pathway of VLA-4 involved activated FAK and Akt, but apparently not PKCα/δ. VLA-4, however, appears representative of other integrins which together impact the heparanase/integrin activation axis in tumorigenicity. Biosensor measurements provided an insight as to how heparin can interfere with this activation process. While low-molecular-weight heparin (LMWH) cannot replace heparanase bound to SDC-4, LMWH can compete with SDC-4 binding of heparanase. Since blockade of heparanase by LMWH has functional consequences for reduced VLA-4 binding, latent heparanase appears as a novel, so far unnoticed target of heparin, underlying its antimetastatic activity.

Keywords

heparanase - heparin - melanoma - cell adhesion - migration - syndecan-4 - VLA-4

• References

• 1 Labelle M, Hynes RO. The initial hours of metastasis: the importance of cooperative host-tumor cell interactions during hematogenous dissemination. Cancer Discov 2012; 2 (12) 1091-1099
• 2 Läubli H, Borsig L. Selectins promote tumor metastasis. Semin Cancer Biol 2010; 20 (3) 169-177
• 3 Chen M, Geng J-G. P-selectin mediates adhesion of leukocytes, platelets, and cancer cells in inflammation, thrombosis, and cancer growth and metastasis. Arch Immunol Ther Exp (Warsz) 2006; 54 (2) 75-84
• 4 Gay LJ, Felding-Habermann B. Contribution of platelets to tumour metastasis. Nat Rev Cancer 2011; 11 (2) 123-134
• 5 Grailer JJ, Kodera M, Steeber DA. L-selectin: role in regulating homeostasis and cutaneous inflammation. J Dermatol Sci 2009; 56 (3) 141-147
• 6 Bendas G, Borsig L. Cancer cell adhesion and metastasis: selectins, integrins, and the inhibitory potential of heparins. Int J Cell Biol 2012; doi: 10.1155/2012/676731
• 7 Schlesinger M, Roblek M, Ortmann K , et al. The role of VLA-4 binding for experimental melanoma metastasis and its inhibition by heparin. Thromb Res 2014; 133 (5) 855-862
• 8 Schadendorf D, Heidel J, Gawlik C, Suter L, Czarnetzki BM. Association with clinical outcome of expression of VLA-4 in primary cutaneous malignant melanoma as well as P-selectin and E-selectin on intratumoral vessels. J Natl Cancer Inst 1995; 87 (5) 366-371
• 9 Schadendorf D, Gawlik C, Haney U, Ostmeier H, Suter L, Czarnetzki BM. Tumour progression and metastatic behaviour in vivo correlates with integrin expression on melanocytic tumours. J Pathol 1993; 170 (4) 429-434
• 10 Okahara H, Yagita H, Miyake K, Okumura K. Involvement of very late activation antigen 4 (VLA-4) and vascular cell adhesion molecule 1 (VCAM-1) in tumor necrosis factor alpha enhancement of experimental metastasis. Cancer Res 1994; 54 (12) 3233-3236
• 11 Garofalo A, Chirivi RG, Foglieni C , et al. Involvement of the very late antigen 4 integrin on melanoma in interleukin 1-augmented experimental metastases. Cancer Res 1995; 55 (2) 414-419
• 12 Higashiyama A, Watanabe H, Okumura K, Yagita H. Involvement of tumor necrosis factor alpha and very late activation antigen 4/vascular cell adhesion molecule 1 interaction in surgical-stress-enhanced experimental metastasis. Cancer Immunol Immunother 1996; 42 (4) 231-236
• 13 Lazo-Langner A, Goss GD, Spaans JN, Rodger MA. The effect of low-molecular-weight heparin on cancer survival. A systematic review and meta-analysis of randomized trials. J Thromb Haemost 2007; 5 (4) 729-737
• 14 Mousa SA, Mohamed S. Inhibition of endothelial cell tube formation by the low molecular weight heparin, tinzaparin, is mediated by tissue factor pathway inhibitor. Thromb Haemost 2004; 92 (3) 627-633
• 15 Goldshmidt O, Zcharia E, Cohen M , et al. Heparanase mediates cell adhesion independent of its enzymatic activity. FASEB J 2003; 17 (9) 1015-1025
• 16 Ramani VC, Yang Y, Ren Y, Nan L, Sanderson RD. Heparanase plays a dual role in driving hepatocyte growth factor (HGF) signaling by enhancing HGF expression and activity. J Biol Chem 2011; 286 (8) 6490-6499
• 17 Thompson CA, Purushothaman A, Ramani VC, Vlodavsky I, Sanderson RD. Heparanase regulates secretion, composition, and function of tumor cell-derived exosomes. J Biol Chem 2013; 288 (14) 10093-10099
• 18 Vlodavsky I, Beckhove P, Lerner I , et al. Significance of heparanase in cancer and inflammation. Cancer Microenviron 2012; 5 (2) 115-132
• 19 SST0001 in Advanced Multiple Myeloma—ClinicalTrials.gov NCT01764880. Available at: http://clinicaltrials.gov/show/NCT01764880 ; Accessed September 11, 2014
• 20 Ostapoff KT, Awasthi N, Cenik BK , et al. PG545, an angiogenesis and heparanase inhibitor, reduces primary tumor growth and metastasis in experimental pancreatic cancer. Mol Cancer Ther 2013; 12 (7) 1190-1201
• 21 Pisano C, Vlodavsky I, Ilan N, Zunino F. The potential of heparanase as a therapeutic target in cancer. Biochem Pharmacol 2014; 89 (1) 12-19
• 22 Gilat D, Hershkoviz R, Goldkorn I , et al. Molecular behavior adapts to context: heparanase functions as an extracellular matrix-degrading enzyme or as a T cell adhesion molecule, depending on the local pH. J Exp Med 1995; 181 (5) 1929-1934
• 23 Zetser A, Bashenko Y, Miao H-Q, Vlodavsky I, Ilan N. Heparanase affects adhesive and tumorigenic potential of human glioma cells. Cancer Res 2003; 63 (22) 7733-7741
• 24 Sotnikov I, Hershkoviz R, Grabovsky V , et al. Enzymatically quiescent heparanase augments T cell interactions with VCAM-1 and extracellular matrix components under versatile dynamic contexts. J Immunol 2004; 172 (9) 5185-5193
• 25 Levy-Adam F, Feld S, Suss-Toby E, Vlodavsky I, Ilan N. Heparanase facilitates cell adhesion and spreading by clustering of cell surface heparan sulfate proteoglycans. PLoS ONE 2008; 3 (6) e2319
• 26 Gingis-Velitski S, Zetser A, Kaplan V , et al. Heparanase uptake is mediated by cell membrane heparan sulfate proteoglycans. J Biol Chem 2004; 279 (42) 44084-44092
• 27 Schlesinger M, Simonis D, Schmitz P, Fritzsche J, Bendas G. Binding between heparin and the integrin VLA-4. Thromb Haemost 2009; 102 (5) 816-822
• 28 Schmitz P, Gerber U, Schütze N , et al. Cyr61 is a target for heparin in reducing MV3 melanoma cell adhesion and migration via the integrin VLA-4. Thromb Haemost 2013; 110 (5) 1046-1054
• 29 Fujita M, Takada YK, Takada Y. The chemokine fractalkine can activate integrins without CX3CR1 through direct binding to a ligand-binding site distinct from the classical RGD-binding site. PLoS ONE 2014; 9 (5) e96372
• 30 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 (13) 12103-12113
• 31 Zetser A, Bashenko Y, Edovitsky E, Levy-Adam F, Vlodavsky I, Ilan N. Heparanase induces vascular endothelial growth factor expression: correlation with p38 phosphorylation levels and Src activation. Cancer Res 2006; 66 (3) 1455-1463
• 32 Vlodavsky I. Preparation of extracellular matrices produced by cultured corneal endothelial and PF-HR9 endodermal cells. Curr Protoc Cell Biol 2001; ; Chapter 10: Unit 10.4
• 33 Liang C-C, Park AY, Guan J-L. In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro. Nat Protoc 2007; 2 (2) 329-333
• 34 Schlesinger M, Schmitz P, Zeisig R , et al. The inhibition of the integrin VLA-4 in MV3 melanoma cell binding by non-anticoagulant heparin derivatives. Thromb Res 2012; 129 (5) 603-610
• 35 Cardones AR, Murakami T, Hwang ST. CXCR4 enhances adhesion of B16 tumor cells to endothelial cells in vitro and in vivo via beta(1) integrin. Cancer Res 2003; 63 (20) 6751-6757
• 36 Couchman JR, Chen L, Woods A. Syndecans and cell adhesion. Int Rev Cytol 2001; 207: 113-150
• 37 Mostafavi-Pour Z, Askari JA, Parkinson SJ, Parker PJ, Ng TTC, Humphries MJ. Integrin-specific signaling pathways controlling focal adhesion formation and cell migration. J Cell Biol 2003; 161 (1) 155-167
• 38 Woods A, Longley RL, Tumova S, Couchman JR. Syndecan-4 binding to the high affinity heparin-binding domain of fibronectin drives focal adhesion formation in fibroblasts. Arch Biochem Biophys 2000; 374 (1) 66-72
• 39 de Bruyn KMT, Rangarajan S, Reedquist KA, Figdor CG, Bos JL. The small GTPase Rap1 is required for Mn(2+)- and antibody-induced LFA-1- and VLA-4-mediated cell adhesion. J Biol Chem 2002; 277 (33) 29468-29476
• 40 Liu S, Thomas SM, Woodside DG , et al. Binding of paxillin to alpha4 integrins modifies integrin-dependent biological responses. Nature 1999; 402 (6762) 676-681
• 41 Webb BA, Chimenti M, Jacobson MP, Barber DL. Dysregulated pH: a perfect storm for cancer progression. Nat Rev Cancer 2011; 11 (9) 671-677
• 42 Meads MB, Gatenby RA, Dalton WS. Environment-mediated drug resistance: a major contributor to minimal residual disease. Nat Rev Cancer 2009; 9 (9) 665-674
• 43 Damiano JS, Cress AE, Hazlehurst LA, Shtil AA, Dalton WS. Cell adhesion mediated drug resistance (CAM-DR): role of integrins and resistance to apoptosis in human myeloma cell lines. Blood 1999; 93 (5) 1658-1667
• 44 Jacamo R, Chen Y, Wang Z , et al. Reciprocal leukemia-stroma VCAM-1/VLA-4-dependent activation of NF-κB mediates chemoresistance. Blood 2014; 123 (17) 2691-2702
• 45 Morgan MR, Humphries MJ, Bass MD. Synergistic control of cell adhesion by integrins and syndecans. Nat Rev Mol Cell Biol 2007; 8 (12) 957-969