title: Nucleoside diphosphate kinase B regulates angiogenic responses in the endothelium via caveolae formation and c-Src-mediated caveolin-1 phosphorylation.
abstract: Nucleoside diphosphate kinase B (NDPK-B) is an enzyme required for nucleoside triphosphate homeostasis, which has been shown to interact with caveolin-1 (Cav-1). In endothelial cells (ECs), NDPK-B contributes to the regulation of angiogenesis and adherens junction (AJ) integrity. We therefore investigated whether an interaction of NDPK-B with Cav-1 in ECs is required for this regulation and the involvement of VEGF signaling herein. We report that simultaneous depletion of NDPK-B/Cav-1 in HUVECs synergistically impaired sprouting angiogenesis. NDPK-B depletion alone impaired caveolae formation, VEGF-induced phosphorylation of c-Src/Cav-1 but not of ERK1/2/AKT/eNOS. In vivo, Cav-1-/-mice showed impaired retinal vascularization at postnatal-day five, whereas NDPK-B-/-mice did not. Primary mouse brain ECs (MBMECs) from NDPK-B-/-mice showed no change in caveolae content and transendothelial-electrical resistance upon VEGF stimulation. Interestingly, NDPK-B-/-MBMECs displayed an accumulation of intracellular vesicles and increased Cav-1 levels. Dextran tracer analysis showed increased vascular permeability in the brain of NDPK-B-/-mice compared to wild type. In conclusion, our data indicate that NDPK-B is required for the correct localization of Cav-1 at the plasma membrane and the formation of caveolae. The genetic ablation of NDPK-B could partially be compensated by an increased Cav-1 content, which restored caveolae formation and some endothelial functions.
authors.affiliation: 1 Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
authors.forename: Shalini
authors.initials: S
authors.lastname: Gross
authors.affiliation: 2 Institute of Neurology (Edinger-Institute), Goethe University, Frankfurt, Germany.
authors.forename: Kavi
authors.initials: K
authors.lastname: Devraj
authors.affiliation: 1 Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
authors.forename: Yuxi
authors.initials: Y
authors.lastname: Feng
authors.affiliation: 2 Institute of Neurology (Edinger-Institute), Goethe University, Frankfurt, Germany.
authors.forename: Jadranka
authors.initials: J
authors.lastname: Macas
authors.affiliation: 2 Institute of Neurology (Edinger-Institute), Goethe University, Frankfurt, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Liebner
authors.affiliation: 1 Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Wieland
mesh_terms: D000818:Animals; D001921:Brain / Q000098:blood supply; D000081247:CSK Tyrosine-Protein Kinase; D021941:Caveolae / Q000378:metabolism* / Q000648:ultrastructure; D051242:Caveolin 1 / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D002463:Cell Membrane Permeability; D042783:Endothelial Cells; D004730:Endothelium, Vascular / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D061307:Human Umbilical Vein Endothelial Cells; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008854:Microscopy, Electron; D055806:Microvessels / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D054778:NM23 Nucleoside Diphosphate Kinases / Q000235:genetics / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000235:genetics / Q000502:physiology*; D010766:Phosphorylation; D012171:Retinal Vessels / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D051242:Caveolin 1; D054778:NM23 Nucleoside Diphosphate Kinases; D042461:Vascular Endothelial Growth Factor A; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human
keywords: Angiogenesis; basic science; blood–brain barrier; endothelium; vascular biology