title: Heterogeneous susceptibility of valve endothelial cells to mesenchymal transformation in response to TNFα.
abstract: Lack of understanding of the early mechanisms of aortic valve stenosis and calcification hinders the development of diagnostic and therapeutic intervention strategies. Inflammation is a known component of early aortic valve disease and can induce mesenchymal transformation in a subset of aortic valve endothelial cells. Here we present a three-dimensional culture system that allows transforming and non-transforming cells to be independently isolated and analyzed. We have used the system to identify and characterize the dynamic invasion and phenotypic transition of two distinct subsets of endothelial cells: those that invade and transform under TNFα treatment, and those that resist mesenchymal transformation and remain endothelial. We determine that non-transformed cells maintain control levels of endothelial genes VE-cadherin and eNOS, while transformed cells lose these endothelial characteristics and upregulate α-smooth muscle actin. Both subsets of cells have an inflammatory phenotype marked by increased ICAM-1, but transformed cells have increased MMP-9, Notch1, TGF-β, and BMP-4, while non-transformed cells do not. Transformed cells also have distinct effects on alignment of collagen fibers as they invade the hydrogel system, which is not found in control endothelial or interstitial valve cells. Understanding the role of transforming and non-transforming endothelial cells in valve disease will provide an important pathological link between early inflammation and later stages of disease. Discovery of the molecular signature of transformation-resistant endothelial cells could inform development of treatment strategies that promote survival of the valve endothelium.
authors: Department of Biomedical Engineering, Cornell University, 304 Weill Hall, Ithaca, NY, 14853, USA.
authors: Emily J
authors: EJ
authors: Farrar
authors: Jonathan T
authors: JT
authors: Butcher
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis; D001021:Aortic Valve / Q000378:metabolism* / Q000473:pathology; D015820:Cadherins / Q000096:biosynthesis; D054338:Cell Transdifferentiation / Q000187:drug effects*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism* / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis; D051881:Receptor, Notch1 / Q000096:biosynthesis; D013552:Swine; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000494:pharmacology*; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D018799:Intercellular Adhesion Molecule-1; D020780:Matrix Metalloproteinase 9; D051881:Receptor, Notch1; D016212:Transforming Growth Factor beta; D014409:Tumor Necrosis Factor-alpha; D000099152:Cadherin 5
references: J Clin Invest. 2009 Jun;119(6):1420-8
references: 19487818
references: Cardiovasc Pathol. 2000 Sep-Oct;9(5):281-6
references: 11064275
references: Arterioscler Thromb Vasc Biol. 2010 Feb;30(2):225-31
references: 19926833
references: J Clin Invest. 1993 Apr;91(4):1800-9
references: 8473518
references: Circulation. 2013 Jan 1;127(1):e6-e245
references: 23239837
references: Cancer Res. 2013 Mar 15;73(6):1981-92
references: 23378344
references: J Vis Exp. 2010 Dec 29;(46):
references: 21248687
references: Arterioscler Thromb Vasc Biol. 2011 Mar;31(3):598-607
references: 21164078
references: Eur Heart J. 2010 Aug;31(16):1975-84
references: 20601388
references: Circ Res. 2005 Apr 15;96(7):792-9
references: 15761200
references: Tissue Eng. 2006 Apr;12(4):905-15
references: 16674302
references: Am J Pathol. 2001 Oct;159(4):1335-43
references: 11583961
references: Fibrogenesis Tissue Repair. 2011 Apr 06;4:11
references: 21470408
references: Eur Heart J. 2005 Jul;26(13):1333-41
references: 15827062
references: Biochem Biophys Res Commun. 2008 Sep 26;374(3):512-6
references: 18647596
references: Circulation. 2007 Jan 23;115(3):377-86
references: 17224478
references: J Cardiovasc Transl Res. 2011 Dec;4(6):710-9
references: 21789724
references: Proc Natl Acad Sci U S A. 2011 Dec 13;108(50):19943-8
references: 22123981
references: Nature. 2005 Sep 8;437(7056):270-4
references: 16025100
references: Arterioscler Thromb Vasc Biol. 2009 Feb;29(2):254-60
references: 19023092
references: Nat Cell Biol. 2000 Feb;2(2):76-83
references: 10655586
references: Am J Pathol. 2008 Nov;173(5):1275-85
references: 18832581
references: J Am Coll Cardiol. 2007 Sep 25;50(13):1205-13
references: 17888836
references: Dev Biol. 2007 Nov 1;311(1):172-84
references: 17916348
references: PLoS One. 2011;6(6):e20969
references: 21698246
references: Circ Res. 2004 Aug 6;95(3):253-60
references: 15217906
references: Arterioscler Thromb Vasc Biol. 2013 Jan;33(1):121-30
references: 23104848
references: Am J Pathol. 2013 May;182(5):1922-31
references: 23499458
references: Dev Cell. 2011 Aug 16;21(2):288-300
references: 21839921
references: J Heart Valve Dis. 1999 May;8(3):254-60
references: 10399657
references: J Am Coll Cardiol. 2000 Dec;36(7):2257-62
references: 11127470
references: Ann Thorac Surg. 2003 Feb;75(2):457-65; discussion 465-6
references: 12607654