The functional role of blood platelet components in angiogenesis

Summary

The process of neovascularization greatly depends on the induction of the angiogenic phenotype of endothelial cells that is strictly controlled by humoral factors as well as by cellular communications in the vascular system. Although blood platelets contain several secretable pro-and antiangiogenic components, their overall role in angiogenesis remains poorly understood. In a mouse model of hypoxia-induced retinal angiogenesis, the situation of thrombocytopenia as well as inhibition of platelet aggregation by a highly specific αIIbß3-integrin antagonist or acetyl salicylic acid (Aspirin™) administration, respectively, resulted in about 35-50% reduction of retinal neovascularization, compatible with a significant contribution of blood platelets in angiogenesis. Platelet remnants and microvesicles were found at sites of angiogenic sprouts. In vitro isolated platelets incorporated in a fibrin gel induced capillary sprouting of microvascular endothelial cells. Similarly, platelet releasate elevated the permeability of confluent endothelial cell monolayers to the same extent as hypoxia did. Platelet-derivedVEGF as well as butanol-extractable lipid mediators were identified as predominant activators of angiogenesis, particularly of microvascular endothelial cell proliferation and migration. In addition, a synergistic effect between platelet-derived VEGF and bFGF in capillary sprouting and endothelial cell proliferation was found. Based on this proangiogenic role of platelets in neovascularization, anti-platelet substances can be considered as potent inhibitors of angiogenesis.

Keywords

Vascular endothelial growth factor - retinal neovascularization - lysophospholipids - thrombocytopenia - anti-platelet drugs

^* These authors contributed equally to this work.

• References

• 1 Cines DB, Pollak ES, Buck CA. et al. Endothelial cells in physiology and in the pathophysiology of vascular disorders. Blood 1998; 91: 3527-3561.
• 2 Ferrara N. Role of vascular endothelial growth factor in regulation of physiological angiogenesis. Am J Physiol Cell Physiol 2001; 280: C1358-66.
• 3 Verheul HM, Jorna AS, Hoekman K. et al. Vascular endothelial growth factor-stimulated endothelial cells promote adhesion and activation of platelets. Blood 2000; 96: 4216-4221.
• 4 Lu M, Perez VL, Ma N, Miyamoto K. et al. VEGF increases retinal vascular ICAM-1 expression in vivo. Invest Ophthalmol Vis Sci 1999; 40: 1808-1812.
• 5 Verheul HM, Hoekman K, Luykx-de Bakker S. et al. Platelets: transporter of vascular endothelial growth factor. Clin Cancer Res 1997; 03: 2187-2190.
• 6 English D, Garcia JG, Brindley DN. Plateletreleased phospholipids link haemostasis and angiogenesis. Cardiovasc Res 2001; 49: 588-599.
• 7 Hla T, Lee MJ, Ancellin N. et al. Lysophospholipids-receptor revelations. Science 2001; 294: 1875-1878.
• 8 May AE, Neumann FJ, Preissner KT. The relevance of blood cell-vessel wall adhesive interactions for vascular thrombotic disease. Thromb Haemost 1999; 82: 962-970.
• 9 Pinedo HM, Verheul HM, D’Amato RJ. et al. Involvement of platelets in tumour angiogenesis?. Lancet 1988; 352: 1775-1777.
• 10 Iruela-Arispe ML, Lombardo M, Krutzsch HC. et al. Inhibition of angiogenesis by thrombospondin-1 is mediated by 2 independent regions within the type 1 repeats. Circulation 1999; 100: 1423-1431.
• 11 Jouan V, Canron X, Alemany M. et al. Inhibition of in vitro angiogenesis by platelet factor-4-derived peptides and mechanism of action. Blood 1999; 94: 984-993.
• 12 Pepper MS, Montesano R, Vassalli JD. et al. Chondrocytes inhibit endothelial sprout formation in vitro: evidence for involvement of a transforming growth factor-ß. J Cell Physiol 1999; 146: 170-179.
• 13 Browder T, Folkman J, Pirie-Shepherd S. The hemostatic system as a regulator of angiogenesis. J Biol Chem 2000; 275: 1521-1524.
• 14 Chen J, Bierhaus A, Schiekofer S. et al. Tissue factor – a receptor involved in the control of cellular properties, including angiogenesis. Thromb Haemost 2001; 86: 334-345.
• 15 Haralabopoulos GC, Grant DS, Kleinman HK. et al. Thrombin promotes endothelial cell alignment in Matrigel in vitro and angiogenesis in vivo. Am J Physiol 1997; 273: C239-45.
• 16 van Hinsbergh VW, Collen A, Koolwijk P. Role of fibrin matrix in angiogenesis. Ann N Y Acad Sci 2001; 936: 426-437.
• 17 Hejna M, Raderer M, Zielinski CC. Inhibition of metastases by anticoagulants. J Natl Cancer Inst 1999; 91: 22-36.
• 18 Koolwijk P, van Erck MG, de Vree WJ. et al. Cooperative effect of TNFα, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity. J Cell Biol 1996; 132: 1177-1188.
• 19 O’Reilly MS, Holmgren L, Shing Y. et al. Angiostatin: a circulating endothelial cell inhibitor that suppresses angiogenesis and tumor growth. Cold Spring Harb Symp Quant Biol 1994; 59: 471-482.
• 20 Colman RW, Jameson BA, Lin Y. et al. Domain 5 of high molecular weight kininogen (kininostatin) down-regulates endothelial cell proliferation and migration and inhibits angiogenesis. Blood 2000; 95: 543-550.
• 21 O’Reilly MS, Pirie-Shepherd S, Lane WS. et al. Antiangiogenic activity of the cleaved conformation of the serpin antithrombin. Science 1999; 285: 1926-1928.
• 22 Hammes HP, Brownlee M, Jonczyk A. et al. Subcutaneous injection of a cyclic peptide antagonist of vitronectin receptor-type integrins inhibits retinal neovascularization. Nature Med 1996; 02: 529-533.
• 23 Chavakis E, Riecke B, Lin J. et al. Kinetics of integrin expression in the mouse model of proliferative retinopathy and success of secondary intervention with cyclic RGD peptides. Diabetologia 2002; 45: 262-267.
• 24 Laschinger M, Engelhardt B. Interaction of α4-integrin with VCAM-1 is involved in adhesion of encephalitogenic T cell blasts to brain endothelium but not in their transendothelial migration in vitro. J Neuroimmunol 2000; 102: 32-43.
• 25 Jaffe EA, Nachman RL, Becker CG. et al. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 1973; 52: 2745-2756.
• 26 Mischeck U, Meyer J, Galla HJ. Characterization of γ-glutamyl transpeptidase activity of cultured endothelial cells from porcine brain capillaries. Cell Tissue Res 1989; 256: 221-226.
• 27 Wohn KD, Schmidt T, Kanse SM. et al. The role of plasminogen activator inhibitor-1 as inhibitor of platelet and megakaryoblastic cell adhesion. Br J Haematol 1999; 104: 901-908.
• 28 Smith LE, Wesolowski E, McLellan A. et al. Oxygen-induced retinopathy in the mouse. Invest Ophthalmol Vis Sci 1994; 35: 101-111.
• 29 Mahalingam M, Ugen KE, Kao KJ. et al. Functional role of platelets in experimental metastasis studied with cloned murine fibrosarcoma cell variants. Cancer Res 1988; 48: 1460-1464.
• 30 Stoelcker B, Hafner M, Orosz P. et al. Role of adhesion molecules and platelets in TNFinduced adhesion of tumor cells to endothelial cells: implications for experimental metastasis. J Inflamm 1995; 46: 155-167.
• 31 Graham RC, Karnovsky MJ. The histochemical demonstration of uricase activity. J Histochem Cytochem 1965; 13: 448-453.
• 32 Hynes RO. A reevaluation of integrins as regulators of angiogenesis. Nature Med 2002; 08: 918-921.
• 33 Denis C, Methia N, Frenette PS. et al. A mouse model of severe von Willebrand disease: defects in hemostasis and thrombosis. Proc Natl Acad Sci USA 1998; 95: 9524-9529.
• 34 Hynes RO, Hodivala-Dilke KM. Insights and questions arising from studies of a mouse model of Glanzmann thrombasthenia. Thromb Haemost 1999; 82: 481-485.
• 35 Hodivala-Dilke KM, McHugh KP, Tsakiris DA. et al. ß3-integrin-deficient mice are a model for Glanzmann thrombasthenia showing placental defects and reduced survival. J Clin Invest 1999; 103: 229-238.
• 36 Cheresh DA, Stupack DG. Integrin-mediated death: an explanation of the integrin-knockout phenotype?. Nature Med 2002; 08: 193-194.
• 37 Ohlmann P, Eckly A, Freund M. et al. ADP induces partial platelet aggregation without shape change and potentiates collageninduced aggregation in the absence of Gαq. Blood 96: 2000; 2134-2139.
• 38 Mohle R, Green D, Moore MA. et al. Constitutive production and thrombin-induced release of vascular endothelial growth factor by human megakaryocytes and platelets. Proc Natl Acad Sci USA 1997; 94: 663-668.
• 39 Goto F, Goto K, Weindel K. et al. Synergistic effects of vascular endothelial growth factor and basic fibroblast growth factor on the proliferation and cord formation of bovine capillary endothelial cells within collagen gels. Lab Invest 1993; 69: 508-517.
• 40 Asahara T, Bauters C, Zheng LP. et al. Synergistic effect of vascular endothelial growth factor and basic fibroblast growth factor on angiogenesis in vivo . Circulation 1995; 92: 365-371.
• 41 Pintucci G, Froum S, Pinnell J. et al. Trophic effects of platelets on cultured endothelial cells are mediated by platelet-associated fibroblast growth factor-2 (FGF-2) and vascular endothelial growth factor (VEGF). Thromb Haemost 2002; 88: 834-842.
• 42 Friedlander M, Brooks PC, Shaffer RW. et al. Definition of two angiogenic pathways by distinct αv integrins. Science 1995; 270: 1500-1502.
• 43 English D, Welch Z, Kovala AT. et al. Sphingosine 1-phosphate released from platelets during clotting accounts for the potent endothelial cell chemotactic activity of blood serum and provides a novel link between hemostasis and angiogenesis. FASEB J 2000; 14: 2255-2265.
• 44 Senger DR, Van de Water L, Brown LF. et al. Vascular permeability factor (VPF, VEGF) in tumor biology. Cancer Metastasis Rev 1993; 12: 303-324.
• 45 Fischer S, Renz D, Schaper W. et al. Effects of barbiturates on hypoxic cultures of brain derived microvascular endothelial cells. Brain Res 1996; 707: 47-53.
• 46 Manegold PC, Hutter J, Pahernik SA. et al. Platelet-endothelial interaction in tumor angiogenesis and microcirculation. Blood 2003; 101: 1970-1976.
• 47 Müller I, Klocke A, Alex M. et al. Intravascular tissue factor initiates coagulation via circulating microvesicles and platelets. FASEB J 2003; 17: 476-478.
• 48 Kim HK, Song KS, Park YS. et al. Elevated levels of circulating platelet microparticles, VEGF, IL-6 and RANTES in patients with gastric cancer: possible role of a metastasis predictor Eur J Cancer. 2003; 39: 184-191.
• 49 Shimada H, Takeda A, Nabeya Y. et al. Clinical significance of serum vascular endothelial growth factor in esophageal squamous cell carcinoma. Cancer 2001; 92: 663-669.
• 50 Hyodo I, Doi T, Endo H. et al. Clinical significance of plasma vascular endothelial growth factor in gastro-intestinal cancer. Eur J Cancer 1998; 34: 2041-2045.
• 51 Banks RE, Forbes MA, Kinsey SE. et al. Release of the angiogenic cytokine vascular endothelial growth factor (VEGF) from platelets: significance for VEGF measurements and cancer biology”. Br J Cancer 1998; 77: 956-964.
• 52 Amirkhosravi A, Mousa SA, Amaya M. et al. Inhibition of tumor cell-induced platelet aggregation and lung metastasis by the oral GpIIb/IIIa antagonist XV454. Thromb Haemost 2003; 90: 549-554.
• 53 Varner JA, Nakada MT, Jordan RE. et al. Inhibition of angiogenesis and tumor growth by murine 7E3, the parent antibody of c7E3 Fab (abciximab; ReoProTM). Angiogenesis 1999; 03: 53-60.
• 54 Xu Y, Shen Z, Wiper DW, Wu M, Morton RE, Elson P, Kennedy AW, Belinson J, Markman M, Casey G. Lysophosphatidic acid as a potential biomarker for ovarian and other gynecologic cancers. JAMA 1998; 280: 719-723.
• 55 Ma L, Elliott SN, Cirino G. et al. Platelets modulate gastric ulcer healing: Role of endostatin and vascular endothelial growth factor release. Proc Natl Acad Sci USA 2001; 98: 6470-6475.
• 56 Verheul HMW, Panigrahy D, Yuan J. et al. Combination oral antiangiogenic therapy with thalidomide and sulindac inhibits tumour growth in rabbits. Br J Cancer 1999; 79: 114-118.
• 57 Smorenburg SM, Hettiarachchi RJ, Vink R. et al. The effects of unfractionated heparin on survival in patients with malignancy – a systematic review. Thromb Haemost 1999; 82: 1600-1604.