title: Novel role of p66Shc in ROS-dependent VEGF signaling and angiogenesis in endothelial cells.
abstract: p66Shc, a longevity adaptor protein, is demonstrated as a key regulator of reactive oxygen species (ROS) metabolism involved in aging and cardiovascular diseases. Vascular endothelial growth factor (VEGF) stimulates endothelial cell (EC) migration and proliferation primarily through the VEGF receptor-2 (VEGFR2). We have shown that ROS derived from Rac1-dependent NADPH oxidase are involved in VEGFR2 autophosphorylation and angiogenic-related responses in ECs. However, a role of p66Shc in VEGF signaling and physiological responses in ECs is unknown. Here we show that VEGF promotes p66Shc phosphorylation at Ser36 through the JNK/ERK or PKC pathway as well as Rac1 binding to a nonphosphorylated form of p66Shc in ECs. Depletion of endogenous p66Shc with short interfering RNA inhibits VEGF-induced Rac1 activity and ROS production. Fractionation of caveolin-enriched lipid raft demonstrates that p66Shc plays a critical role in VEGFR2 phosphorylation in caveolae/lipid rafts as well as downstream p38MAP kinase activation. This in turn stimulates VEGF-induced EC migration, proliferation, and capillary-like tube formation. These studies uncover a novel role of p66Shc as a positive regulator for ROS-dependent VEGFR2 signaling linked to angiogenesis in ECs and suggest p66Shc as a potential therapeutic target for various angiogenesis-dependent diseases.
authors.affiliation: Department of Pharmacology, Center for Lung and Vascular Biology, University of Illinois, Chicago, IL 60612, USA.
authors.forename: Jin
authors.initials: J
authors.lastname: Oshikawa
authors.forename: Seok-Jo
authors.initials: SJ
authors.lastname: Kim
authors.forename: Eiji
authors.initials: E
authors.lastname: Furuta
authors.forename: Cristiana
authors.initials: C
authors.lastname: Caliceti
authors.forename: Gin-Fu
authors.initials: GF
authors.lastname: Chen
authors.forename: Ronald D
authors.initials: RD
authors.lastname: McKinney
authors.forename: Frank
authors.initials: F
authors.lastname: Kuhr
authors.forename: Irena
authors.initials: I
authors.lastname: Levitan
authors.forename: Tohru
authors.initials: T
authors.lastname: Fukai
authors.forename: Masuko
authors.initials: M
authors.lastname: Ushio-Fukai
mesh_terms: D021941:Caveolae / Q000201:enzymology; D042783:Endothelial Cells / Q000166:cytology / Q000201:enzymology*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D020935:MAP Kinase Signaling System / Q000502:physiology*; D021962:Membrane Microdomains / Q000201:enzymology; D018919:Neovascularization, Physiologic / Q000502:physiology*; D010766:Phosphorylation / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*; D055765:Shc Signaling Adaptor Proteins / Q000378:metabolism*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism
chemical_list: C107953:RAC1 protein, human; D017382:Reactive Oxygen Species; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D040301:Vascular Endothelial Growth Factor Receptor-2; D048051:p38 Mitogen-Activated Protein Kinases; D020830:rac1 GTP-Binding Protein
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