title: VEGF-A Regulates Cellular Localization of SR-BI as Well as Transendothelial Transport of HDL but Not LDL.
abstract: Low- and high-density lipoproteins (LDL and HDL) must pass the endothelial layer to exert pro- and antiatherogenic activities, respectively, within the vascular wall. However, the rate-limiting factors that mediate transendothelial transport of lipoproteins are yet little known. Therefore, we performed a high-throughput screen with kinase drug inhibitors to identify modulators of transendothelial LDL and HDL transport.
  
  
  
  Microscopy-based high-content screening was performed by incubating human aortic endothelial cells with 141 kinase-inhibiting drugs and fluorescent-labeled LDL or HDL. Inhibitors of vascular endothelial growth factor (VEGF) receptors (VEGFR) significantly decreased the uptake of HDL but not LDL. Silencing of VEGF receptor 2 significantly decreased cellular binding, association, and transendothelial transport of125I-HDL but not125I-LDL. RNA interference with VEGF receptor 1 or VEGF receptor 3 had no effect. Binding, uptake, and transport of HDL but not LDL were strongly reduced in the absence of VEGF-A from the cell culture medium and were restored by the addition of VEGF-A. The restoring effect of VEGF-A on endothelial binding, uptake, and transport of HDL was abrogated by pharmacological inhibition of phosphatidyl-inositol 3 kinase/protein kinase B or p38 mitogen-activated protein kinase, as well as silencing of scavenger receptor BI. Moreover, the presence of VEGF-A was found to be a prerequisite for the localization of scavenger receptor BI in the plasma membrane of endothelial cells.
  
  
  
  The identification of VEGF as a regulatory factor of transendothelial transport of HDL but not LDL supports the concept that the endothelium is a specific and, hence, druggable barrier for the entry of lipoproteins into the vascular wall.
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Srividya
authors.initials: S
authors.lastname: Velagapudi
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Mustafa
authors.initials: M
authors.lastname: Yalcinkaya
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Antonio
authors.initials: A
authors.lastname: Piemontese
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Roger
authors.initials: R
authors.lastname: Meier
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Simon Flyvbjerg
authors.initials: SF
authors.lastname: Nørrelykke
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Damir
authors.initials: D
authors.lastname: Perisa
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Andrzej
authors.initials: A
authors.lastname: Rzepiela
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Michael
authors.initials: M
authors.lastname: Stebler
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Szymon
authors.initials: S
authors.lastname: Stoma
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.).
authors.forename: Paolo
authors.initials: P
authors.lastname: Zanoni
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.). arnold.voneckardstein@usz.ch lucia.rohrer@usz.ch.
authors.forename: Lucia
authors.initials: L
authors.lastname: Rohrer
authors.affiliation: From the Institute of Clinical Chemistry, University and University Hospital of Zurich, Schlieren, Switzerland (S.V., M.Y., A.P., D.P., P.Z., L.R., A.v.E.); Competence Center for Integrated Human Physiology, University of Zurich, Switzerland (S.V., M.Y., D.P., P.Z., L.R., A.v.E.); Department of Pharmacy, University of Parma, Italy (A.P.); and Scientific Center for Optical and Electron Microscopy, ETH Zurich, Switzerland (R.M., S.F.N., A.R., M.S., S.S.). arnold.voneckardstein@usz.ch lucia.rohrer@usz.ch.
authors.forename: Arnold
authors.initials: A
authors.lastname: von Eckardstein
mesh_terms: D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D057166:High-Throughput Screening Assays / Q000379:methods; D006801:Humans; D008075:Lipoproteins, HDL / Q000378:metabolism*; D008077:Lipoproteins, LDL / Q000378:metabolism*; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D021381:Protein Transport; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D051122:Scavenger Receptors, Class B / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
chemical_list: D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; C495124:SCARB1 protein, human; D051122:Scavenger Receptors, Class B; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D058539:Phosphatidylinositol 3-Kinase; C000588866:KDR protein, human; D040301:Vascular Endothelial Growth Factor Receptor-2; D051057:Proto-Oncogene Proteins c-akt; D048051:p38 Mitogen-Activated Protein Kinases
keywords: HDL; SR-BI; VEGF-A; atherosclerosis; endothelial cells