Tissue factor pathway inhibitor (TFPI) interferes with endothelial cell migration by inhibition of both the Erk pathway and focal adhesion proteins

Summary

Tissue factor pathway inhibitor (TFPI) is a plasma Kunitz-type serine protease inhibitor that is mainly known for its inhibition of tissue factor-mediated coagulation. In addition to its anticoagulant properties, emerging data show that TFPI may also regulate endothelial cell functions via a non-haemostatic pathway. In this work we demonstrate that at concentrations within the physiological range,TFPI inhibits both endothelial cell migration and their differentiation into capillary-like structures in vitro. These effects were specific to endothelial cells since no inhibitory effect was observed on the migration of tumor (glio- blastoma) cells. Inhibition of endothelial cell migration was correlated with a concomitant loss in cell adhesion,suggesting an alteration of focal adhesion complex integrity. Accordingly,we observed thatTFPI inhibited the phosphorylation of focal adhesion kinase and paxillin,two key proteins involved in the scaffolding of these complexes, and that this effect was specific to endothelial cells. These results suggest that TFPI influences the angiogenic process via a non-haemostatic pathway, by downregulating the migratory mechanisms of endothelial cells.

Keywords

Angiogenesis - TFPI - endothelial cell migration - focal adhesion complexes - extracellular signal-regulated kinases

• References

• 1 Cavallaro U, Christofori G. Molecular mechanism of tumor angiogenesis and tumor progression. J Neurooncol 2000; 50: 63-70.
• 2 Bergers G, Benjamin LE. Tumorigenesis and the angiogenic switch. Nat Rev Cancer 2003; 3: 401-410.
• 3 Carmeliet P. Angiogenesis in health and disease. Nat Med 2003; 9: 653-660.
• 4 Nijziel MR, Van Oerle R, Hillen HFP. et al. From Trousseau to angiogenesis: the link between haemosatic system and cancer. Neth J Med 2006; 64: 403-410.
• 5 Wojtukiewicz MZ, Sierko E, Rak J. Contribution of the hemostatic system to angiogenesis in cancer. Semin Thromb Hemost 2004; 30: 5-20.
• 6 Belting M, Dorrell MI, Sandgren S. et al. Regulation of angiogenesis by tissue factor cytoplasmic domain signaling. Nat Med 2004; 10: 502-509.
• 7 Lopez-Pedrara C, Barbarroja N, Dorado G. et al. Tissue factor as an effector of angiogenesis and tumor progression in haematological malignancies. Leukemia 2006; 20: 1331-1340.
• 8 Lee OH, Kim YM, Lee MY. Sphingosine-1-phosphate induces angiogenesis: its angiogenic action and signaling mechanism in human umbilical vein endothelial cell. Biochem Biophys Res Commun 1999; 264: 743-750.
• 9 Ozaki H, Hla T, Lee MJ. Sphingosine-1-phosphate signaling in endothelial activation. J Atheroscler Thromb 2003; 10: 125-131.
• 10 Hla T. Signaling and biological actions of sphingosine- 1-phosphate. Pharmacol Res 2003; 47: 401-407.
• 11 Wojtukiewicz MZ, Sierko E, Klement P. et al. The hemostatic system and angiogenesis in malignancy. Neoplasia 2001; 3: 371-384.
• 12 Boccacio C, Medico E. Cancer and blood coagulation. Cell Mol Life Sci 2006; 63: 1024-1027.
• 13 Lwaleed B, Bass PS. Tissue factor pathway inhibitor: structure biology and involvement in disease. J Pathol 2006; 208: 327-339.
• 14 Bajaj MS, Birktoft JJ, Steer SA. et al. Structure and biology of tissue factor pathway inhibitor. Thromb Haemost 2001; 86: 959-972.
• 15 Pedersen B, Holscher T, Sato Y. A balance between tissue factor and tissue factor pathway inhibitor is required for embryonic development and hemostasis in adult mice. Blood 2005; 105: 2777-2782.
• 16 Amirkhosravi A, Meyer T, Chang JY. et al. Tissue factor pathway inhibitor reduces experimental lung metastasis of B16 melanoma. Thromb Haemost 2002; 87: 930-936.
• 17 Hembrough TA, Ruiz JF, Swerdlow BM. et al. Identification and characterization of a very low density lipoprotein receptor-binding peptide from tissue factor pathway inhibitor that has antitumor and antiangiogenic activity. Blood 2004; 103: 3374-3380.
• 18 Gustafson ME, Junger KD, Wun TC. et al. Renaturation and purification of human tissue factor pathway inhibitor expressed in recombinant E. coli. . Protein Expr Purif 1994; 5: 233-241.
• 19 Langlois S, Gingras D, Beliveau R. Membrane type 1-matrix metalloproteinase (MT1-MMP) cooperates with sphingosine 1-phosphate to induce endothelial cell migration and morphogenic differentiation. Blood 2004; 103: 3020-3028.
• 20 Pilorget A, Berthet V, Luis J. et al. Medulloblastoma cell invasion is inhibited by green tea (-)epigallocatechin- 3-gallate. J Cell Biochem 2003; 90: 745-755.
• 21 Mousa SA. Anticoagulants in thrombosis and cancer: the missing link. Sem Thromb Hemost 2002; 28: 45-52.
• 22 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: 627-633.
• 23 Paik JH, Chae SS, Lee MJ. et al. Sphingosine- 1-phosphate-induced endothelial cell migration requires the expression of Edg-1 ad Edg-3 receptorsand Rho-dependent activation of αvβ3 and β1-containing integrins. J Biol Chem 2001; 276: 11830-11837.
• 24 Huttenlocher A, Ginsberg MH, Horwitz AF. Modulation of cell migration by integrin-mediated cytoskeletal linkages and ligand-binding affinity. J Cell Biol 1996; 134: 1551-1562.
• 25 Palacek SP, Loftus JC, Ginsberg MH. et al. Integrin- ligand binding properties govern cell migration speed through cell-substratum adhesiveness. Nature 1997; 385: 537-540.
• 26 Huang C, Jacobson K, Schaller D. Map kinases and cell migration. J Cell Science 2004; 117: 4619-4628.
• 27 Mitra SK, Hanson DA, Schaepfer DD. Focal adhesion kinase in command and control of cell motility. Nat Rev Mol Cell Biol 2005; 6: 56-68.
• 28 Wickström SA, Veikola T, Rehn M. et al. Endostatin- induced modulation of plasminogen activation with concomitant loss of focal adhesions and actin stress fibers in cultured human endothelial cells. Cancer Res 2001; 61: 6511-6516.
• 29 Rapaport SI. Inhibition of factor VIIa/tissue factorinduced coagulation: with particular emphasis upon a factor Xa-dependent mechanism. Blood 1989; 73: 359-365.
• 30 Rapaport SI, Rao LV. The tissue factor pathway: how it has become a “prima ballerina”. Thromb Haemost 1995; 74: 7-17.
• 31 Hembrough TA, Ruiz JF, Papathanassiu AE. Tissue factor pathway inhibitor inhibits endothelial cell proliferation via association with the very low density lipoprotein receptor. J Biol Chem 2001; 276: 12241-12248.
• 32 Guan M, Jin J, Su B. et al. Tissue factor expression and angiogenesis in human glioma. Clin Biochem 2002; 35: 321-325.
• 33 Hamada K, Kuratsu J, Saitoh Y. et al. Expression of tissue factor correlates with grade of malignancy in human glioma. Cancer 1996; 77: 1877-1883.
• 34 Lupu C, Goodwin CA, Westmuckett AD. et al. Tissue factor pathway inhibitor in endothelial cells colocalizes with glycolipid microdomains/caveolae. Regulatory mechanism(s) of the anticoagulant properties of the endothelium. Arterioscler Thromb Vasc Biol 1997; 17: 2964-2974.
• 35 Sevinski JR, Rao Mohan Vijay L, Ruf W. Ligandinduced protease receptor translocation into caveolae: a mechanism for regulating cell surface proteolysis of tissue factor-dependent coagulation pathway. J Cell Biol 1996; 133: 293-304.
• 36 McLean GW, Carragher NO, Avizienyte E. et al. The role of focal adhesion kinase in cancer-A new therapeutic opportunity. Nat Rev Cancer 2005; 5: 505-515.