title: Laminar shear stress inhibits endothelial cell metabolism via KLF2-mediated repression of PFKFB3.
abstract: OBJECTIVE
  Cellular metabolism was recently shown to regulate endothelial cell phenotype profoundly. Whether the atheroprotective biomechanical stimulus elicited by laminar shear stress modulates endothelial cell metabolism is not known.
  
  
  RESULTS
  Here, we show that laminar flow exposure reduced glucose uptake and mitochondrial content in endothelium. Shear stress-mediated reduction of endothelial metabolism was reversed by silencing the flow-sensitive transcription factor Krüppel-like factor 2 (KLF2). Endothelial-specific deletion of KLF2 in mice induced glucose uptake in endothelial cells of perfused hearts. KLF2 overexpression recapitulates the inhibitory effects on endothelial glycolysis elicited by laminar flow, as measured by Seahorse flux analysis and glucose uptake measurements. RNA sequencing showed that shear stress reduced the expression of key glycolytic enzymes, such as 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase-3 (PFKFB3), phosphofructokinase-1, and hexokinase 2 in a KLF2-dependent manner. Moreover, KLF2 represses PFKFB3 promoter activity. PFKFB3 knockdown reduced glycolysis, and overexpression increased glycolysis and partially reversed the KLF2-mediated reduction in glycolysis. Furthermore, PFKFB3 overexpression reversed KLF2-mediated reduction in angiogenic sprouting and network formation.
  
  
  CONCLUSIONS
  Our data demonstrate that shear stress-mediated repression of endothelial cell metabolism via KLF2 and PFKFB3 controls endothelial cell phenotype.
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Anuradha
authors.initials: A
authors.lastname: Doddaballapur
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Katharina M
authors.initials: KM
authors.lastname: Michalik
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Yosif
authors.initials: Y
authors.lastname: Manavski
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Tina
authors.initials: T
authors.lastname: Lucas
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Riekelt H
authors.initials: RH
authors.lastname: Houtkooper
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Xintian
authors.initials: X
authors.lastname: You
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Andreas M
authors.initials: AM
authors.lastname: Zeiher
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Michael
authors.initials: M
authors.lastname: Potente
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.).
authors.forename: Stefanie
authors.initials: S
authors.lastname: Dimmeler
authors.affiliation: From the Institute for Cardiovascular Regeneration, Center of Molecular Medicine, Goethe University, Frankfurt am Main, Germany (A.D., K.M.M., Y.M., T.L., S.D., R.A.B.); The Laboratory Genetic Metabolic Diseases, Academic Medical Center, Amsterdam, The Netherlands (R.H.H.); The Max-Delbrück-Center, Berlin, Germany (X.Y., W.C.); Department of Cardiology, Internal Medicine III, Goethe University Hospital Frankfurt, Frankfurt am Main, Germany (A.M.Z.); Angiogenesis and Metabolism Laboratory, Max Planck Institute for Heart and Lung Research, Bad Nauheim, Germany (M.P.); and German Center for Cardiovascular Research (DZHK), Partner Site RheinMain, Frankfurt am Main, Germany (A.M.Z., S.D.). boon@med.uni-frankfurt.de.
authors.forename: Reinier A
authors.initials: RA
authors.lastname: Boon
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002478:Cells, Cultured; D015536:Down-Regulation; D042783:Endothelial Cells / Q000201:enzymology*; D004734:Energy Metabolism; D005947:Glucose / Q000378:metabolism; D006019:Glycolysis; D061307:Human Umbilical Vein Endothelial Cells / Q000201:enzymology; D051741:Kruppel-Like Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D040542:Mechanotransduction, Cellular; D018345:Mice, Knockout; D008928:Mitochondria / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D018919:Neovascularization, Physiologic; D010641:Phenotype; D025481:Phosphofructokinase-2 / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012039:Regional Blood Flow; D013314:Stress, Mechanical; D013997:Time Factors; D014162:Transfection
chemical_list: C096161:KLF2 protein, human; C498314:Klf2 protein, mouse; D051741:Kruppel-Like Transcription Factors; C107345:PFKFB3 protein, human; C000595376:PFKFB3 protein, mouse; D025481:Phosphofructokinase-2; D005947:Glucose
keywords: angiogenesis; endothelium; glycolysis; hemodynamics; metabolism; shear stress down regulated gene-1 protein, human