HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Membrane type 1–matrix metalloproteinase (MT1-MMP) cooperates with sphingosine 1–phosphate to induce endothelial cell migration and morphogenic differentiation

Membrane type 1–matrix metalloproteinase (MT1-MMP) has been suggested to play an important role in angiogenesis, but the mechanisms involved remain incompletely understood. Using an in vitro model of angiogenesis in which cell migration of bovine aortic endothelial cells (BAECs) and their morphogenic differentiation into capillary-like structures on Matrigel are induced by overexpression of MT1-MMP, we show that the plateletderived bioactive lipid sphingosine 1–phosphate (S1P) is the predominant serum factor essential for MT1-MMP– dependent migration and morphogenic differentiation activities. In the presence of S1P, MT1-MMP–dependent cell migration and morphogenic differentiation were inhibited by pertussis toxin, suggesting the involvement of Gi-protein–coupled receptor-mediated signaling. Accordingly, cotransfection of BAECs with MT1-MMP and a constitutively active G i2 (Q205L) mutant increased cell migration and morphogenic differentiation, whereas treatment of BAECs overexpressing MT1MMP with antisense oligonucleotides directed against S1P1 and S1P3, the predominant S1P receptors, significantly inhibited both processes. These results demonstrate that MT1-MMP–induced migration and morphogenic differentiation involve the cooperation of the enzyme with platelet-derived bioactive lipids through S1P-mediated activation of G icoupled S1P1 and S1P3 receptors. Given the important contribution of platelets to tumor angiogenesis, the stimulation of endothelial MT1-MMP function by S1P may thus constitute an important molecular event linking hemostasis to angiogenesis. (Blood. 2004;103:3020-3028)