{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T04:09:01Z","timestamp":1792642141865,"version":"4.5.0"},"reference-count":45,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2021,3,2]],"date-time":"2021-03-02T00:00:00Z","timestamp":1614643200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01HL126705, R01HL145064, R01HL145064-01S1"],"award-info":[{"award-number":["R01HL126705, R01HL145064, R01HL145064-01S1"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100011410","name":"School of Medicine, University of South Carolina","doi-asserted-by":"publisher","award":["Bob Price - Instrumentation Resource Facility Endowment Fund"],"award-info":[{"award-number":["Bob Price - Instrumentation Resource Facility Endowment Fund"]}],"id":[{"id":"10.13039\/100011410","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JCDD"],"abstract":"<jats:p>Among the three transforming growth factor beta (TGF\u03b2) ligands, TGF\u03b22 is essential for heart development and is produced by multiple cell types, including myocardium. Heterozygous mutations in TGFB2 in patients of connective tissue disorders result in congenital heart defects and adult valve malformations, including mitral valve prolapse (MVP) with or without regurgitation. Tgfb2 germline knockout fetuses exhibit multiple cardiac defects but the role of myocardial-TGF\u03b22 in heart development is yet to be elucidated. Here, myocardial Tgfb2 conditional knockout (CKO) embryos were generated by crossing Tgfb2flox mice with Tgfb2+\/\u2212; cTntCre mice. Tgfb2flox\/\u2212 embryos were normal, viable. Cell fate mapping was done using dual-fluorescent mT\/mG+\/\u2212 mice. Cre-mediated Tgfb2 deletion was assessed by genomic PCR. RNAscope in situ hybridization was used to detect the loss of myocardial Tgfb2 expression. Histological, morphometric, immunohistochemical, and in situ hybridization analyses of CKOs and littermate controls at different stages of heart development (E12.5\u2013E18.5) were used to determine the role of myocardium-derived TGF\u03b22 in atrioventricular (AV) cushion remodeling and myocardial development. CKOs exhibit a thin ventricular myocardium, AV cushion remodeling defects and developed incomplete AV septation defects. The loss of myocardial Tgfb2 resulted in impaired cushion maturation and dysregulated cell death. Phosphorylated SMAD2, a surrogate for TGF\u03b2 signaling, was \u201cparadoxically\u201d increased in both AV cushion mesenchyme and ventricular myocardium in the CKOs. Our results indicate that TGF\u03b22 produced by cardiomyocytes acting as cells autonomously on myocardium and via paracrine signaling on AV cushions are required for heart development.<\/jats:p>","DOI":"10.3390\/jcdd8030026","type":"journal-article","created":{"date-parts":[[2021,3,2]],"date-time":"2021-03-02T10:36:37Z","timestamp":1614681397000},"page":"26","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Myocardial TGF\u03b22 Is Required for Atrioventricular Cushion Remodeling and Myocardial Development"],"prefix":"10.3390","volume":"8","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6479-3981","authenticated-orcid":false,"given":"Aniket","family":"Bhattacharya","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadia","family":"Al-Sammarraie","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1513-9844","authenticated-orcid":false,"given":"Mengistu G.","family":"Gebere","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Johnson","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1053-9045","authenticated-orcid":false,"given":"John F.","family":"Eberth","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9228-8096","authenticated-orcid":false,"given":"Mohamad","family":"Azhar","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Anatomy, University of South Carolina School of Medicine, Columbia, SC 29209, USA"},{"name":"William Jennings Bryan Dorn VA Medical Center, Dorn Research Institute, Columbia, SC 29209, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,3,2]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"eaav5183","DOI":"10.1126\/scisignal.aav5183","article-title":"Specificity, versatility, and control of TGF-beta family signaling","volume":"12","author":"Derynck","year":"2019","journal-title":"Sci. Signal."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1111\/cga.12048","article-title":"Epicardium is required for sarcomeric maturation and cardiomyocyte growth in the ventricular compact layer mediated by transforming growth factor beta and fibroblast growth factor before the onset of coronary circulation","volume":"54","author":"Takahashi","year":"2014","journal-title":"Congenit. Anom. Kyoto"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1214","DOI":"10.1161\/CIRCRESAHA.118.314438","article-title":"Protective Effects of Activated Myofibroblasts in the Pressure-Overloaded Myocardium Are Mediated Through Smad-Dependent Activation of a Matrix-Preserving Program","volume":"124","author":"Russo","year":"2019","journal-title":"Circ. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"S52","DOI":"10.21037\/jtd.2016.11.19","article-title":"The role of transforming growth factor (TGF)-beta in the infarcted myocardium","volume":"9","author":"Frangogiannis","year":"2017","journal-title":"J. Thorac. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1007\/s00441-011-1241-3","article-title":"Transforming growth factor beta signaling in adult cardiovascular diseases and repair","volume":"347","author":"Doetschman","year":"2012","journal-title":"Cell Tissue Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1002\/dvdy.10314","article-title":"Expression patterns of Tgfbeta1-3 associate with myocardialisation of the outflow tract and the development of the epicardium and the fibrous heart skeleton","volume":"227","author":"Molin","year":"2003","journal-title":"Dev. Dyn."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"916","DOI":"10.1038\/ng.2348","article-title":"TGFB2 mutations cause familial thoracic aortic aneurysms and dissections associated with mild systemic features of Marfan syndrome","volume":"44","author":"Boileau","year":"2012","journal-title":"Nat. Genet."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"922","DOI":"10.1038\/ng.2349","article-title":"Loss-of-function mutations in TGFB2 cause a syndromic presentation of thoracic aortic aneurysm","volume":"44","author":"Lindsay","year":"2012","journal-title":"Nat. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1111\/j.1749-6632.2009.05098.x","article-title":"Developmental basis of adult cardiovascular diseases: Valvular heart diseases","volume":"1188","author":"Markwald","year":"2010","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1038\/nature14670","article-title":"Mutations in DCHS1 cause mitral valve prolapse","volume":"525","author":"Durst","year":"2015","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.clp.2015.11.002","article-title":"Genetic and Developmental Basis of Cardiovascular Malformations","volume":"43","author":"Azhar","year":"2016","journal-title":"Clin. Perinatol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1002\/dvdy.21854","article-title":"Ligand-specific function of transforming growth factor beta in epithelial-mesenchymal transition in heart development","volume":"238","author":"Azhar","year":"2009","journal-title":"Dev. Dyn."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1002\/dvdy.22702","article-title":"Transforming growth factor Beta2 is required for valve remodeling during heart development","volume":"240","author":"Azhar","year":"2011","journal-title":"Dev. Dyn."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.tcm.2005.09.006","article-title":"Transforming growth factor beta-SMAD2 signaling and aortic arch development","volume":"16","author":"Azhar","year":"2006","journal-title":"Trends Cardiovasc. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1242\/dev.098632","article-title":"High-resolution imaging of cardiomyocyte behavior reveals two distinct steps in ventricular trabeculation","volume":"141","author":"Staudt","year":"2014","journal-title":"Development"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1159\/000146455","article-title":"Morphologic study of ventricular trabeculation in the embryonic chick heart","volume":"130","author":"Icardo","year":"1987","journal-title":"Acta Anat. (Basel)"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1038\/s41586-018-0110-6","article-title":"Control of cardiac jelly dynamics by NOTCH1 and NRG1 defines the building plan for trabeculation","volume":"557","author":"Ramialison","year":"2018","journal-title":"Nature"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1387\/ijdb.130302bk","article-title":"Remodeling of the myocardium in early trabeculation and cardiac valve formation; a role for TGFbeta2","volume":"57","author":"Kruithof","year":"2013","journal-title":"Int. J. Dev. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1136\/hrt.2005.082271","article-title":"Isolated ventricular non-compaction of the myocardium in adults","volume":"93","author":"Jenni","year":"2007","journal-title":"Heart"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1002\/dvg.22795","article-title":"Generation of mice carrying a knockout-first and conditional-ready allele of transforming growth factor beta2 gene","volume":"52","author":"Bose","year":"2014","journal-title":"Genesis"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1161\/01.RES.73.5.783","article-title":"Transforming growth factor-beta in cardiac ontogeny and adaptation","volume":"73","author":"MacLellan","year":"1993","journal-title":"Circ. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2745","DOI":"10.1161\/01.CIR.103.22.2745","article-title":"Double-outlet right ventricle and overriding tricuspid valve reflect disturbances of looping, myocardialization, endocardial cushion differentiation, and apoptosis in TGF-beta(2)-knockout mice","volume":"103","author":"Bartram","year":"2001","journal-title":"Circulation"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"van den Hoff, M.J.B., and Wessels, A. (2020). Muscularization of the Mesenchymal Outlet Septum during Cardiac Development. J. Cardiovasc. Dev. Dis., 7.","DOI":"10.3390\/jcdd7040051"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2362","DOI":"10.1101\/gad.1124803","article-title":"An essential role of Bmp4 in the atrioventricular septation of the mouse heart","volume":"17","author":"Jiao","year":"2003","journal-title":"Genes Dev."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1242\/dev.124.13.2659","article-title":"TGFbeta2 knockout mice have multiple developmental defects that are non-overlapping with other TGFbeta knockout phenotypes","volume":"124","author":"Sanford","year":"1997","journal-title":"Development"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"593","DOI":"10.1002\/dvg.20335","article-title":"A global double-fluorescent Cre reporter mouse","volume":"45","author":"Muzumdar","year":"2007","journal-title":"Genesis"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"200","DOI":"10.1016\/j.ydbio.2008.01.003","article-title":"Periostin regulates atrioventricular valve maturation","volume":"316","author":"Norris","year":"2008","journal-title":"Dev. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"616","DOI":"10.1038\/nrm3434","article-title":"TGFbeta signalling in context","volume":"13","author":"Massague","year":"2012","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1609","DOI":"10.1038\/onc.2012.191","article-title":"Genome-wide mechanisms of Smad binding","volume":"32","author":"Morikawa","year":"2013","journal-title":"Oncogene"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1080\/1354750X.2016.1217932","article-title":"Genetic variance of transforming growth factor beta2 gene in conotruncal heart defects","volume":"22","author":"Chen","year":"2017","journal-title":"Biomarkers"},{"key":"ref_31","first-page":"210","article-title":"Association between rs6658835 polymorphism of transforming growth factor beta 2 gene and congenital heart diseases in Chinese Han population","volume":"29","author":"Xie","year":"2012","journal-title":"Zhonghua Yi Xue Yi Chuan Xue Za Zhi"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"4332","DOI":"10.1093\/eurheartj\/ehaa156","article-title":"Atrial myxomas arise from multipotent cardiac stem cells","volume":"41","author":"Scalise","year":"2020","journal-title":"Eur. Heart J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1016\/S1359-6101(03)00044-3","article-title":"Transforming growth factor beta in cardiovascular development and function","volume":"14","author":"Azhar","year":"2003","journal-title":"Cytokine Growth Factor Rev."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"659","DOI":"10.1172\/JCI123547","article-title":"Lineage-specific events underlie aortic root aneurysm pathogenesis in Loeys-Dietz syndrome","volume":"129","author":"Macfarlane","year":"2019","journal-title":"J. Clin. Investig."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2878","DOI":"10.1073\/pnas.042390499","article-title":"Endocardial cushion and myocardial defects after cardiac myocyte-specific conditional deletion of the bone morphogenetic protein receptor ALK3","volume":"99","author":"Gaussin","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Burns, T., Yang, Y., Hiriart, E., and Wessels, A. (2016). The Dorsal Mesenchymal Protrusion and the Pathogenesis of Atrioventricular Septal Defects. J. Cardiovasc. Dev. Dis., 3.","DOI":"10.3390\/jcdd3040029"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1080\/15384101.2018.1464852","article-title":"Molecular basis of functional myogenic specification of Bona Fide multipotent adult cardiac stem cells","volume":"17","author":"Cianflone","year":"2018","journal-title":"Cell Cycle."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.ydbio.2005.08.041","article-title":"The right ventricle, outflow tract, and ventricular septum comprise a restricted expression domain within the secondary\/anterior heart field","volume":"287","author":"Verzi","year":"2005","journal-title":"Dev. Biol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"208","DOI":"10.1016\/j.ydbio.2008.07.038","article-title":"Signaling via the Tgf-beta type I receptor Alk5 in heart development","volume":"322","author":"Sridurongrit","year":"2008","journal-title":"Dev. Biol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4585","DOI":"10.1242\/dev.02597","article-title":"Tgfbeta signaling is required for atrioventricular cushion mesenchyme remodeling during in vivo cardiac development","volume":"133","author":"Jiao","year":"2006","journal-title":"Development"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"e34110","DOI":"10.7554\/eLife.34110","article-title":"A novel source of arterial valve cells linked to bicuspid aortic valve without raphe in mice","volume":"7","author":"Eley","year":"2018","journal-title":"eLife"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1005","DOI":"10.1002\/dvdy.24641","article-title":"Intercalated cushion cells within the cardiac outflow tract are derived from the myocardial troponin T type 2 (Tnnt2) Cre lineage","volume":"247","author":"Mifflin","year":"2018","journal-title":"Dev. Dyn."},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Chakrabarti, M., Al-Sammarraie, N., Gebere, M.G., Bhattacharya, A., Chopra, S., Johnson, J., Pe\u00f1a, E.A., Eberth, J.F., Poelmann, R.E., and Groot, A.C.G.-D. (2020). Transforming Growth Factor Beta3 is Required for Cardiovascular Development. J. Cardiovasc. Dev. Dis., 7.","DOI":"10.3390\/jcdd7020019"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1586","DOI":"10.1172\/JCI200422715","article-title":"TGF-beta-dependent pathogenesis of mitral valve prolapse in a mouse model of Marfan syndrome","volume":"114","author":"Ng","year":"2004","journal-title":"J. Clin. Investig."},{"key":"ref_45","unstructured":"Adam, M.P., Ardinger, H.H., Pagon, R.A., Wallace, S.E., Bean, L.J.H., Ghayda Mirzaa, G., and Anne Amemiya, A. (1993). Loeys-Dietz Syndrome. GeneReviews\u00ae, University of Washington."}],"container-title":["Journal of Cardiovascular Development and Disease"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2308-3425\/8\/3\/26\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:31:31Z","timestamp":1760160691000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2308-3425\/8\/3\/26"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,3,2]]},"references-count":45,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["jcdd8030026"],"URL":"https:\/\/doi.org\/10.3390\/jcdd8030026","relation":{},"ISSN":["2308-3425"],"issn-type":[{"value":"2308-3425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,3,2]]}}}