Receptor protein tyrosine phosphatase beta/zeta is a functional binding partner for vascular endothelial growth factor

• Κύριοι συγγραφείς: Koutsioumpa, Marina, Poimenidi, Evangelia, Pantazaka, Evangelia, Theodoropoulou, Christina, Skoura, Angeliki, Megalooikonomou, Vasileios, Kieffer, Nelly, Courty, Jose, Mizumoto, Shuji, Sugahara, Kazuyuki, Papadimitriou, Evangelia
• Άλλοι συγγραφείς: Κουτσιούμπα, Μαρίνα
• Μορφή: Journal (paper)
• Γλώσσα: English
• Έκδοση: 2017
• Θέματα: Chondroitin sulphate; Endothelial cells; Migration; Pleiotrophin; Tyrosine phosphatases; Vascular endothelial growth factor; Θειική χονδροϊτίνη; Ενδοθηλιακά κύτταρα; Μετανάστευση; Πλειοτροφίνη; Φωσφατάσες τυροσίνης; Αγγειακός ενδοθηλιακός αυξητικός παράγοντας

Background: Receptor protein tyrosine phosphatase beta/zeta (RPTPβ/ζ) is a chondroitin sulphate (CS) transmembrane protein tyrosine phosphatase and is a receptor for pleiotrophin (PTN). RPTPβ/ζ interacts with ανβ3 on the cell surface and upon binding of PTN leads to c-Src dephosphorylation at Tyr530, β3 Tyr773 phosphorylation, cell surface nucleolin (NCL) localization and stimulation of cell migration. c-Src-mediated β3 Tyr773 phosphorylation is also observed after vascular endothelial growth factor 165 (VEGF165) stimulation of endothelial cells and is essential for VEGF receptor type 2 (VEGFR2) - ανβ3 integrin association and subsequent signaling. In the present work, we studied whether RPTPβ/ζ mediates angiogenic actions of VEGF. Methods: Human umbilical vein endothelial, human glioma U87MG and stably transfected Chinese hamster ovary cells expressing different β3 subunits were used. Protein-protein interactions were studied by a combination of immunoprecipitation/Western blot, immunofluorescence and proximity ligation assays, properly quantified as needed. RPTPβ/ζ expression was down-regulated using small interference RNA technology. Migration assays were performed in 24-well microchemotaxis chambers, using uncoated polycarbonate membranes with 8 μm pores. Results: RPTPβ/ζ mediates VEGF165-induced c-Src-dependent β3 Tyr773 phosphorylation, which is required for VEGFR2-ανβ3 interaction and the downstream activation of phosphatidylinositol 3-kinase (PI3K) and cell surface NCL localization. RPTPβ/ζ directly interacts with VEGF165, and this interaction is not affected by bevacizumab, while it is interrupted by both CS-E and PTN. Down-regulation of RPTPβ/ζ by siRNA or administration of exogenous CS-E abolishes VEGF165-induced endothelial cell migration, while PTN inhibits the migratory effect of VEGF165 to the levels of its own effect. Conclusions: These data identify RPTPβ/ζ as a cell membrane binding partner for VEGF that regulates angiogenic functions of endothelial cells and suggest that it warrants further validation as a potential target for development of additive or alternative anti-VEGF therapies.