title: Tetraspanin CD63 promotes vascular endothelial growth factor receptor 2-β1 integrin complex formation, thereby regulating activation and downstream signaling in endothelial cells in vitro and in vivo.
abstract: CD63 is a member of the transmembrane-4 glycoprotein superfamily (tetraspanins) implicated in the regulation of membrane protein trafficking, leukocyte recruitment, and adhesion processes. We have investigated the involvement of CD63 in endothelial cell (EC) signaling downstream of β1 integrin and VEGF. We report that silencing of CD63 in primary ECs arrested capillary sprouting and tube formation in vitro because of impaired adhesion and migration of ECs. Mechanistically, CD63 associated with both β1 integrin and the main VEGF receptor on ECs, VEGFR2. Our data suggest that CD63 serves to bridge between β1 integrin and VEGFR2 because CD63 silencing disrupted VEGFR2-β1 integrin complex formation identified using proximity ligation assays. Signaling downstream of β1 integrin and VEGFR2 was attenuated in CD63-silenced cells, although their cell surface expression levels remained unaffected. CD63 was furthermore required for efficient internalization of VEGFR2 in response to VEGF. Importantly, systemic delivery of VEGF failed to potently induce VEGFR2 phosphorylation and downstream signaling in CD63-deficient mouse lungs. Taken together, our findings demonstrate a previously unrecognized role for CD63 in coordinated integrin and receptor tyrosine kinase signaling in vitro and in vivo.
authors: Department of Immunology, Genetics, and Pathology, Rudbeck Laboratory, Uppsala University, 75185 Uppsala, Sweden. tugues@immunology.uzh.ch
authors: Sònia
authors: S
authors: Tugues
authors: Satoshi
authors: S
authors: Honjo
authors: Christian
authors: C
authors: König
authors: Narendra
authors: N
authors: Padhan
authors: Jeffrey
authors: J
authors: Kroon
authors: Laura
authors: L
authors: Gualandi
authors: Xiujuan
authors: X
authors: Li
authors: Irmeli
authors: I
authors: Barkefors
authors: Victor L
authors: VL
authors: Thijssen
authors: Arjan W
authors: AW
authors: Griffioen
authors: Lena
authors: L
authors: Claesson-Welsh
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002462:Cell Membrane / Q000378:metabolism; D002633:Chemotaxis; D042783:Endothelial Cells / Q000166:cytology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism*; D008168:Lung / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009389:Neovascularization, Pathologic; D011485:Protein Binding; D034741:RNA, Small Interfering / Q000378:metabolism; D015398:Signal Transduction; D060149:Tetraspanin 30 / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*
chemical_list: D019012:Integrin beta1; D034741:RNA, Small Interfering; D060149:Tetraspanin 30; D040301:Vascular Endothelial Growth Factor Receptor-2
keywords: Angiogenesis; CD63; Endothelial Cell; Receptor Tyrosine Kinase; Tetraspanins; Vascular Endothelial Growth Factor (VEGF)
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