title: Valve Endothelial Cell-Derived Tgfβ1 Signaling Promotes Nuclear Localization of Sox9 in Interstitial Cells Associated With Attenuated Calcification.
abstract: OBJECTIVE
  Aortic valve disease, including calcification, affects >2% of the human population and is caused by complex interactions between multiple risk factors, including genetic mutations, the environment, and biomechanics. At present, there are no effective treatments other than surgery, and this is because of the limited understanding of the mechanisms that underlie the condition. Previous work has shown that valve interstitial cells within the aortic valve cusps differentiate toward an osteoblast-like cell and deposit bone-like matrix that leads to leaflet stiffening and calcific aortic valve stenosis. However, the mechanisms that promote pathological phenotypes in valve interstitial cells are unknown.
  
  
  RESULTS
  Using a combination of in vitro and in vivo tools with mouse, porcine, and human tissue, we show that in valve interstitial cells, reduced Sox9 expression and nuclear localization precedes the onset of calcification. In vitro, Sox9 nuclear export and calcific nodule formation is prevented by valve endothelial cells. However, in vivo, loss of Tgfβ1 in the endothelium leads to reduced Sox9 expression and calcific aortic valve disease.
  
  
  CONCLUSIONS
  Together, these findings suggest that reduced nuclear localization of Sox9 in valve interstitial cells is an early indicator of calcification, and therefore, pharmacological targeting to prevent nuclear export could serve as a novel therapeutic tool in the prevention of calcification and stenosis.
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: Danielle J
authors.initials: DJ
authors.lastname: Huk
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: Blair F
authors.initials: BF
authors.lastname: Austin
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: Tori E
authors.initials: TE
authors.lastname: Horne
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: Robert B
authors.initials: RB
authors.lastname: Hinton
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: William C
authors.initials: WC
authors.lastname: Ray
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.).
authors.forename: Donald D
authors.initials: DD
authors.lastname: Heistad
authors.affiliation: From the Molecular and Cellular Pharmacology Graduate Program, Leonard M. Miller School of Medicine, Miami, FL (D.J.H.); Center for Cardiovascular Research and The Heart Center at Nationwide Children's Hospital Research Institute, Columbus, OH (D.J.H., B.F.A., T.E.H., J.L.); Division of Cardiology, The Heart Institute, Cincinnati Children's Hospital Medical Center, Cincinnati, OH (R.B.H.); Battelle Center for Mathematical Medicine, Nationwide Children's Hospital Research Institute, Columbus, OH (W.C.R.); The Ohio State University Interdisciplinary Graduate Program in Biophysics, Columbus, OH (W.C.R.); Department of Pediatrics, The Ohio State University, Columbus, OH (W.C.R., J.L.); and Division of Cardiovascular Medicine and Department of Pharmacology, University of Iowa Carver College of Medicine, Iowa City, IA (D.D.H.). joy.lincoln@nationwidechildrens.org.
authors.forename: Joy
authors.initials: J
authors.lastname: Lincoln
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D001021:Aortic Valve / Q000378:metabolism* / Q000473:pathology*; D001024:Aortic Valve Stenosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000517:prevention & control; D002114:Calcinosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000517:prevention & control; D002478:Cells, Cultured; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D019899:Paracrine Communication; D055758:SOX9 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013552:Swine; D013997:Time Factors; D046509:Tissue Culture Techniques; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000235:genetics / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000378:metabolism
chemical_list: C447413:Col2a1 protein, mouse; D024043:Collagen Type II; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; C527933:Sox9 protein, mouse; C508960:TGFB1 protein, human; C508961:Tgfb1 protein, mouse; D053773:Transforming Growth Factor beta1; D054460:rho-Associated Kinases
keywords: animal model cardiovascular disease; aortic valve; endothelial cell; heart valve; signaling pathways