Data from Dissecting the Role of Matrix Metalloproteinases (MMP) and Integrin α<sub>v</sub>β<sub>3</sub> in Angiogenesis <i>In vitro</i>: Absence of Hemopexin C Domain Bioactivity, but Membrane-Type 1-MMP and α<sub>v</sub>β<sub>3</sub> Are Critical

Matrix metalloproteinase (MMP)-2 and its hemopexin C domain autolytic fragment (also called PEX) have been proposed to be crucial for angiogenesis. Here, we have investigated the dependency of in vitro angiogenesis on MMP-mediated extracellular proteolysis and integrin α_vβ₃–mediated cell adhesion in a three-dimensional collagen I model. The hydroxamate-based synthetic inhibitors BB94, CT1399, and CT1847 inhibited endothelial cell invasion, as did neutralizing anti–membrane-type 1-MMP (MT1-MMP) antibodies and tissue inhibitor of MMP (TIMP)-2 and TIMP-3 but not TIMP-1. This confirmed the pivotal importance of MT1-MMP over other MMPs in this model. Invasion was also inhibited by a nonpeptidic antagonist of integrin α_vβ₃, EMD 361276. Although PEX strongly inhibited pro-MMP-2 activation, when contaminating lipopolysaccharide was neutralized, PEX neither affected angiogenesis nor bound integrin α_vβ₃. Moreover, no specific binding of pro-MMP-2 to integrin α_vβ₃ was found, whereas only one out of four independently prepared enzymatically active MMP-2 preparations could bind integrin α_vβ₃, and this in a PEX-independent manner. Likewise, integrin α_vβ₃–expressing cells did not bind MMP-2-coated surfaces. Hence, these findings show that endothelial cell invasion of collagen I gels is MT1-MMP and α_vβ₃- dependent but MMP-2 independent and does not support a role for PEX in α_vβ₃ integrin binding or in modulating angiogenesis in this system.