Integrin signaling is critical for pathological angiogenesis.
The process of postnatal angiogenesis plays a crucial role in pathogenesis of numerous diseases, including but not limited to tumor growth/metastasis, diabetic retinopathy, and in tissue remodeling upon injury. However, the molecular events underlying this complex process are not well understood and numerous issues remain controversial, including the regulatory function of integrin receptors. To analyze the role of integrin phosphorylation and signaling in angiogenesis, we generated knock-in mice that express a mutant beta3 integrin unable to undergo tyrosine phosphorylation. Two distinct models of pathological angiogenesis revealed that neovascularization is impaired in mutant beta3 knock-in mice. In an ex vivo angiogenesis assay, mutant beta3 knock-in endothelial cells did not form complete capillaries in response to vascular endothelial growth factor (VEGF) stimulation. At the cellular level, defective tyrosine phosphorylation in mutant beta3 knock-in cells resulted in impaired adhesion, spreading, and migration of endothelial cells. At the molecular level, VEGF stimulated complex formation between VEGF receptor-2 and beta3 integrin in wild-type but not in mutant beta3 knock-in endothelial cells. Moreover, phosphorylation of VEGF receptor-2 was significantly reduced in cells expressing mutant beta3 compared to wild type, leading to impaired integrin activation in these cells. These findings provide novel mechanistic insights into the role of integrin-VEGF axis in pathological angiogenesis.
Mahabeleshwar
Ganapati H
GH
Department of Molecular Cardiology, Joseph J. Jacobs Center for Thrombosis and Vascular Biology, NB50, The Cleveland Clinic Foundation, Cleveland, OH 44195, USA.
Feng
Weiyi
W
Phillips
David R
DR
Byzova
Tatiana V
TV
D000818:Animals; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism; D039661:Integrin beta3 / Q000235:genetics / Q000502:physiology*; D008546:Melanoma, Experimental; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009389:Neovascularization, Pathologic / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D010766:Phosphorylation; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology*
D039661:Integrin beta3