Regulation of tenascin-C, a vascular smooth muscle cell survival factor that interacts with the α(v)β3 integrin to promote epidermal growth factor receptor phosphorylation and growth

Abstract

Tenascin-C (TN-C) is induced in pulmonary vascular disease, where it colocalizes with proliferating smooth muscle cells (SMCs) and epidermal growth factor (EGF). Furthermore, cultured SMCs require TN-C for EGF- dependent growth on type I collagen. In this study, we explore the regulation and function of TN-C in SMCs. We show that a matix metalloproteinase (MMP) inhibitor (GM6001) suppresses SMC TN-C expression on native collagen, whereas denatured collagen promotes TN-C expression in a β3 integrin dependent manner, independent of MMPs. Floating type I collagen gel also suppresses SMC MMP activity and TN-C protein synthesis and induces apoptosis, in the presence of EGF. Addition of exogenous TN-C to SMCs on floating collagen, or to SMCs treated with GM6001, restores the EGF growth response and 'res- cues' cells from apoptosis. The mechanism by which TN-C facilitates EGF-dependent survival and growth was then investigated. We show that TN-C interactions with α(v)β3 integrins modify SMC shape, and EGF-dependent growth. These features are associated with redistribution of filamentous actin to focal adhesion complexes, which colocalize with dusters of EGF-Rs, tyrosine- phosphorylated proteins, and increased activation of EGF-Rs after addition of EGF. Cross-linking SMC β3 integrins replicates the effect of TN-C on EGF-R clustering and tyrosine phosphorylation. Together, these studies represent a functional paradigm for ECM-dependent cell survival whereby MMPs upregulate TN-C by generating β3 integrin ligands in type I collagen. In turn, α(v)β3 interactions with TN-C alter SMC shape and increase EGF-R clustering and EGF-dependent growth. Conversely, suppression of MMPs downregulates TN-C and induces apoptosis.