Membrane type 1–matrix metalloproteinase (MT1-MMP) cooperates with sphingosine 1–phosphate to induce endothelial cell migration and morphogenic differentiation

Abstract

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-MMPdependent 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 G i -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 MT1-MMP with antisense oligonucleotides directed against S1P 1 and S1P 3 , 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 S1P 1 and S1P 3 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. IntroductionAngiogenesis, the formation of new vessels from pre-existing endothelium, is an essential component of pathologic conditions such as rheumatoid arthritis, diabetic retinopathies, inflammation, arteriosclerosis and tumor growth, and metastasis. 1 Tumor angiogenesis is an extremely complex process in which tumor cells secrete a number of stimulatory cytokines, such as vascular endothelial growth factor (VEGF), 2 that induce proliferation, migration, and survival of the endothelial cells (ECs). 3 VEGF is also a potent inducer of vascular permeability, resulting in the extravasation of plasma fibrinogen in the extravascular space and in the formation of a cross-linked fibrin provisional matrix that is essential for neovascularization. 4 Since new vessels are highly permeable, fibrin is continuously present into the tumor stroma and reflects the activation of the coagulation system, 5 suggesting an important regulatory function of the hemostatic system in angiogenesis and tumor growth. Such a close association between human cancer and hemostasis is illustrated by the observation that cancer coagulopathies occur in more than 80% of patients with disseminated cancer and account for a significant percentage of the morbidity and mortality of this disease. 6,7 The mechanisms involved in the pathogenesis of hemostatic disorders in cancer remains, however, incompletely understood but likely involve alterations in the function of some components ...

Membrane-Type 1 Matrix Metalloproteinase Stimulates Cell Migration through Epidermal Growth Factor Receptor Transactivation

“…These morphologic changes were also inhibited by BB94 (30% of the GFP-positive cells showed a migratory phenotype; Fig. 2), a broad-spectrum metalloprotease inhibitor, which is in keeping with our previous study showing that MT1-MMPdependent BAEC migration involves the catalytic activity of the enzyme (13). In addition, overexpression of a catalytically inactive MT1-MMP mutant (E240A) and treatment with MT1-MMP physiologic tissue inhibitors of metalloproteinase (TIMP-2 and TIMP-3) inhibited the ability of MT1-MMP to induce BAEC migratory phenotype (data not shown).…”

“…1B). Although a close cooperation between the S1P-and the vascular endothelial growth factor -mediated signaling cascades has been identified in some cell systems, we have previously shown that the S1P-stimulated MT1-MMP -dependent endothelial cell migration and morphogenic differentiation do not involve vascular endothelial growth factor receptor activity (13). To determine whether the stimulatory effect of S1P involves the transactivation of other receptor tyrosine kinases, such as PDGFR (24), and EGFR (24, 25), we tested AG1478 and AG1296, two selective inhibitors of EGFR and PDGFR tyrosine kinase activities, respectively.…”

“…Both of these processes involve the activation of G protein -mediated signaling via S1P 1 and S1P 3 receptors (13). Although bovine aortic endothelial cells (BAEC), which express low levels of MT1-MMP, do not spontaneously form capillary-like structures on Matrigel (Fig.…”

“…The short cytoplasmic sequence of MT1-MMP also contributes to the induction of cell migration by the enzyme (9, 10, 13) possibly through activation of the extracellular signal-regulated protein kinase (ERK) cascade (14), endocytosis and trafficking of the enzyme (15,16), and interaction with tyrosine phosphorylated caveolin-1 (17). Additional insights into the mechanisms by which MT1-MMP induces cell migration may come from our observation that MT1-MMP cooperates with the plateletderived bioactive lipid sphingosine 1-phosphate (S1P) to stimulate endothelial cell migration and morphogenic differentiation into capillary-like structures in vitro (13). The mechanisms involved in the interaction between MT1-MMP and S1P remain largely unknown but might play a significant physiologic role given the importance of the predominant S1P receptor (S1P 1 ) in cell migration (18), blood vessel maturation (19), and tumor angiogenesis (20).…”

“…Several lines of evidence suggest that MT1-MMP -dependent proteolysis of both extracellular matrix components (2,3) and cell surface adhesion receptors (4-6) plays an essential role in tumor growth, invasion, and angiogenesis. The overexpression of MT1-MMP enables tumor cell growth in otherwise growthrestrictive, three-dimensional extracellular matrices (7) and promotes cell migration of a number of cancer (8)(9)(10) and endothelial cell lines (11)(12)(13). The short cytoplasmic sequence of MT1-MMP also contributes to the induction of cell migration by the enzyme (9, 10, 13) possibly through activation of the extracellular signal-regulated protein kinase (ERK) cascade (14), endocytosis and trafficking of the enzyme (15,16), and interaction with tyrosine phosphorylated caveolin-1 (17).…”