Optogenetic interrogation of integrin ?V?3 function in endothelial cells.

ABSTRACT: The integrin ?V?3 is reported to promote angiogenesis in some model systems but not in others. Here, we used optogenetics to study the effects of ?V?3 interaction with the intracellular adapter kindlin-2 (Fermt2) on endothelial cell functions potentially relevant to angiogenesis. Because interaction of kindlin-2 with ?V?3 requires the C-terminal three residues of the ?3 cytoplasmic tail (Arg-Gly-Thr; RGT), optogenetic probes LOVpep and ePDZ1 were fused to ?3?RGT-GFP and mCherry-kindlin-2, respectively, and expressed in ?3 integrin-null microvascular endothelial cells. Exposure of the cells to 450?nm (blue) light caused rapid and specific interaction of kindlin-2 with ?V?3 as assessed by immunofluorescence and total internal reflection fluorescence (TIRF) microscopy, and it led to increased endothelial cell migration, podosome formation and angiogenic sprouting. Analyses of kindlin-2 mutants indicated that interaction of kindlin-2 with other kindlin-2 binding partners, including c-Src, actin, integrin-linked kinase and phosphoinositides, were also likely necessary for these endothelial cell responses. Thus, kindlin-2 promotes ?V?3-dependent angiogenic functions of endothelial cells through its simultaneous interactions with ?3 integrin and several other binding partners. Optogenetic approaches should find further use in clarifying spatiotemporal aspects of vascular cell biology.

SUBMITTER: Liao Z 

PROVIDER: S-EPMC5665447 | biostudies-other | 2017 Oct

REPOSITORIES: biostudies-other