title: A heat-shock protein axis regulates VEGFR2 proteolysis, blood vessel development and repair.
abstract: Vascular endothelial growth factor A (VEGF-A) binds to the VEGFR2 receptor tyrosine kinase, regulating endothelial function, vascular physiology and angiogenesis. However, the mechanism underlying VEGFR2 turnover and degradation in this response is unclear. Here, we tested a role for heat-shock proteins in regulating the presentation of VEGFR2 to a degradative pathway. Pharmacological inhibition of HSP90 stimulated VEGFR2 degradation in primary endothelial cells and blocked VEGF-A-stimulated intracellular signaling via VEGFR2. HSP90 inhibition stimulated the formation of a VEGFR2-HSP70 complex. Clathrin-mediated VEGFR2 endocytosis is required for this HSP-linked degradative pathway for targeting VEGFR2 to the endosome-lysosome system. HSP90 perturbation selectively inhibited VEGF-A-stimulated human endothelial cell migration in vitro. A mouse femoral artery model showed that HSP90 inhibition also blocked blood vessel repair in vivo consistent with decreased endothelial regeneration. Depletion of either HSP70 or HSP90 caused defects in blood vessel formation in a transgenic zebrafish model. We conclude that perturbation of the HSP70-HSP90 heat-shock protein axis stimulates degradation of endothelial VEGFR2 and modulates VEGF-A-stimulated intracellular signaling, endothelial cell migration, blood vessel development and repair.
authors.affiliation: Endothelial Cell Biology Unit, School for Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom.
authors.forename: Alexander F
authors.initials: AF
authors.lastname: Bruns
authors.forename: Nadira
authors.initials: N
authors.lastname: Yuldasheva
authors.forename: Antony M
authors.initials: AM
authors.lastname: Latham
authors.forename: Leyuan
authors.initials: L
authors.lastname: Bao
authors.forename: Caroline
authors.initials: C
authors.lastname: Pellet-Many
authors.forename: Paul
authors.initials: P
authors.lastname: Frankel
authors.forename: Sam L
authors.initials: SL
authors.lastname: Stephen
authors.forename: Gareth J
authors.initials: GJ
authors.lastname: Howell
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Wheatcroft
authors.forename: Mark T
authors.initials: MT
authors.lastname: Kearney
authors.forename: Ian C
authors.initials: IC
authors.lastname: Zachary
authors.forename: Sreenivasan
authors.initials: S
authors.lastname: Ponnambalam
mesh_terms: D000818:Animals; D001158:Arteries / Q000187:drug effects / Q000502:physiology; D016227:Benzoquinones / Q000494:pharmacology; D001808:Blood Vessels / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D002966:Clathrin / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D018919:Neovascularization, Physiologic / Q000187:drug effects; D055550:Protein Stability / Q000187:drug effects; D021381:Protein Transport / Q000187:drug effects; D059748:Proteolysis / Q000187:drug effects; D012038:Regeneration / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism*; D014945:Wound Healing / Q000187:drug effects; D015027:Zebrafish
chemical_list: D016227:Benzoquinones; D002966:Clathrin; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D040301:Vascular Endothelial Growth Factor Receptor-2; C001277:geldanamycin