title: Hic-5 Mediates TGFβ-Induced Adhesion in Vascular Smooth Muscle Cells by a Nox4-Dependent Mechanism.
abstract: OBJECTIVE
  Focal adhesions (FAs) link the cytoskeleton to the extracellular matrix and as such play important roles in growth, migration, and contractile properties of vascular smooth muscle cells. Recently, it has been shown that downregulation of Nox4, a transforming growth factor (TGF) β-inducible, hydrogen peroxide (H2O2)-producing enzyme, affects the number of FAs. However, the effectors downstream of Nox4 that mediate FA regulation are unknown. The FA resident protein H2O2-inducible clone (Hic)-5 is H2O2 and TGFβ inducible, and a binding partner of the heat shock protein (Hsp) 27. The objective of this study was to elucidate the mechanism, by which Hic-5 and Hsp27 participate in TGFβ-induced, Nox4-mediated vascular smooth muscle cell adhesion and migration.
  
  
  RESULTS
  Through a combination of molecular biology and biochemistry techniques, we found that TGFβ, by a Nox4-dependent mechanism, induces the expression and interaction of Hic-5 and Hsp27, which is essential for Hic-5 localization to FAs. Importantly, we found that Hic-5 expression is required for the TGFβ-mediated increase in FA number, adhesive forces and migration. Mechanistically, Nox4 downregulation impedes Smad (small body size and mothers against decapentaplegic) signaling by TGFβ, and Hsp27 and Hic-5 upregulation by TGFβ is blocked in small body size and mothers against decapentaplegic 4-deficient cells.
  
  
  CONCLUSIONS
  Hic-5 and Hsp27 are effectors of Nox4 required for TGFβ-stimulated FA formation, adhesion strength and migration in vascular smooth muscle cell.
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Isabel
authors: I
authors: Fernandez
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Abel
authors: A
authors: Martin-Garrido
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Dennis W
authors: DW
authors: Zhou
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Roza E
authors: RE
authors: Clempus
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Bonnie
authors: B
authors: Seidel-Rogol
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Alejandra
authors: A
authors: Valdivia
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Bernard
authors: B
authors: Lassègue
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Andrés J
authors: AJ
authors: García
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.). kgriend@emory.edu.
authors: Kathy K
authors: KK
authors: Griendling
authors: From the Division of Cardiology, Department of Medicine, Emory University, Atlanta, GA (I.F., A.M.-G., R.E.C., B.S.-R., A.V., B.L., K.K.G., A.S.M.); and Woodruff School of Mechanical Engineering and Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta (D.W.Z., A.J.G.).
authors: Alejandra
authors: A
authors: San Martin
mesh_terms: D002448:Cell Adhesion / Q000235:genetics / Q000502:physiology; D002465:Cell Movement / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D022001:Focal Adhesions / Q000235:genetics / Q000502:physiology; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D060588:LIM Domain Proteins / Q000235:genetics / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D055551:HSP27 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; C490351:TGFB1I1 protein, human; D016212:Transforming Growth Factor beta; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498910:NOX4 protein, human
keywords: Hic5 protein; Hsp27 protein; Nox4 protein; cell migration assay; focal adhesions; vascular smooth muscle