{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T17:03:05Z","timestamp":1778864585111,"version":"3.51.4"},"reference-count":34,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T00:00:00Z","timestamp":1706140800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Institutes of Health","award":["K01OD028205"],"award-info":[{"award-number":["K01OD028205"]}]},{"name":"National Institutes of Health","award":["D19CA-325"],"award-info":[{"award-number":["D19CA-325"]}]},{"DOI":"10.13039\/100001250","name":"Morris Animal Foundation","doi-asserted-by":"publisher","award":["K01OD028205"],"award-info":[{"award-number":["K01OD028205"]}],"id":[{"id":"10.13039\/100001250","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100001250","name":"Morris Animal Foundation","doi-asserted-by":"publisher","award":["D19CA-325"],"award-info":[{"award-number":["D19CA-325"]}],"id":[{"id":"10.13039\/100001250","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Mitral valve prolapse (MVP) is a common valvular disease, affecting 2\u20133% of the adult human population and is a degenerative condition. A total of 5\u201310% of the afflicted will develop severe mitral regurgitation, cardiac dysfunction, congestive heart failure, and sudden cardiac death. Naturally occurring myxomatous MVP in dogs closely resembles MVP in humans structurally, and functional consequences are similar. In both species, valvular interstitial cells (VICs) in affected valves exhibit phenotype consistent with activated myofibroblasts with increased alpha-smooth muscle actin (\u03b1SMA) expression. Using VICs collected from normal and MVP-affected valves of dogs, we analyzed the miRNA expression profile of the cells and their associated small extracellular vesicles (sEV) using RNA sequencing to understand the role of non-coding RNAs and sEV in MVP pathogenesis. miR-145 was shown to be upregulated in both the affected VICs and sEV, and overexpression of miR-145 by mimic transfection in quiescent VIC recapitulates the activated myofibroblastic phenotype. Concurrently, KLF4 expression was noted to be suppressed by miR-145, confirming the miR-145\u2014KLF4\u2014\u03b1SMA axis. Targeting this axis may serve as a potential therapy in controlling pathologic abnormalities found in MVP valves.<\/jats:p>","DOI":"10.3390\/ijms25031468","type":"journal-article","created":{"date-parts":[[2024,1,25]],"date-time":"2024-01-25T03:59:42Z","timestamp":1706155182000},"page":"1468","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Defining the Role of the miR-145\u2014KLF4\u2014\u03b1SMA Axis in Mitral Valvular Interstitial Cell Activation in Myxomatous Mitral Valve Prolapse Using the Canine Model"],"prefix":"10.3390","volume":"25","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1281-4250","authenticated-orcid":false,"given":"Vicky K.","family":"Yang","sequence":"first","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole","family":"Moyer","sequence":"additional","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Runzi","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sally Z.","family":"Carnevale","sequence":"additional","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dawn M.","family":"Meola","sequence":"additional","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4711-1667","authenticated-orcid":false,"given":"Sally R.","family":"Robinson","sequence":"additional","affiliation":[{"name":"Department of Clinical Science, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA 01536, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3874-0744","authenticated-orcid":false,"given":"Guoping","family":"Li","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saumya","family":"Das","sequence":"additional","affiliation":[{"name":"Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA 02114, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101724","DOI":"10.1016\/j.cpcardiol.2023.101724","article-title":"Arrhythmogenic Mitral Valve Prolapse and Sudden Cardiac Death: An Update and Current Perspectives","volume":"48","author":"Battaglia","year":"2023","journal-title":"Curr. Probl. Cardiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1077788","DOI":"10.3389\/fcvm.2023.1077788","article-title":"Genetics and pathophysiology of mitral valve prolapse","volume":"10","author":"Delwarde","year":"2023","journal-title":"Front. Cardiovasc. Med."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Alenazy, A., Eltayeb, A., Alotaibi, M.K., Anwar, M.K., Mulafikh, N., Aladmawi, M., and Vriz, O. (2022). Diagnosis of Mitral Valve Prolapse: Much More than Simple Prolapse. Multimodality Approach to Risk Stratification and Therapeutic Management. J. Clin. Med., 11.","DOI":"10.3390\/jcm11020455"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1016\/j.jacc.2008.05.007","article-title":"2008 focused update incorporated into the ACC\/AHA 2006 guidelines for the management of patients with valvular heart disease: A report of the American College of Cardiology\/American Heart Association Task Force on Practice Guidelines (Writing Committee to revise the 1998 guidelines for the management of patients with valvular heart disease). Endorsed by the Society of Cardiovascular Anesthesiologists, Society for Cardiovascular Angiography and Interventions, and Society of Thoracic Surgeons","volume":"52","author":"Bonow","year":"2008","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1206","DOI":"10.1038\/ng.3383","article-title":"Genetic association analyses highlight biological pathways underlying mitral valve prolapse","volume":"47","author":"Dina","year":"2015","journal-title":"Nat. Genet."},{"key":"ref_6","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_7","first-page":"233","article-title":"MVP-Associated Filamin A Mutations Affect FlnA-PTPN12 (PTP-PEST) Interactions","volume":"2","author":"Duval","year":"2015","journal-title":"J. Cardiovasc. Dev. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.jvc.2012.01.011","article-title":"Historical review, epidemiology and natural history of degenerative mitral valve disease","volume":"14","author":"Borgarelli","year":"2012","journal-title":"J. Vet. Cardiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1111\/j.1748-5827.1974.tb06529.x","article-title":"Observations on the effect of age on the severity of heart valve lesions in the dog","volume":"15","author":"Whitney","year":"1974","journal-title":"J. Small Anim. Pract."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.jvc.2012.01.005","article-title":"Pathology, protein expression and signaling in myxomatous mitral valve degeneration: Comparison of dogs and humans","volume":"14","author":"Aupperle","year":"2012","journal-title":"J. Vet. Cardiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/j.rvsc.2009.01.009","article-title":"Immunohistochemical characterization of the extracellular matrix in normal mitral valves and in chronic valve disease (endocardiosis) in dogs","volume":"87","author":"Aupperle","year":"2009","journal-title":"Res. Vet. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"451","DOI":"10.1042\/bj2190451","article-title":"Collagen composition of normal and myxomatous human mitral heart valves","volume":"219","author":"Cole","year":"1984","journal-title":"Biochem. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1093\/cvr\/cvt083","article-title":"TGF-beta signalling and reactive oxygen species drive fibrosis and matrix remodelling in myxomatous mitral valves","volume":"99","author":"Hagler","year":"2013","journal-title":"Cardiovasc. Res."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Yang, V.K., Tai, A.K., Huh, T.P., Meola, D.M., Juhr, C.M., Robinson, N.A., and Hoffman, A.M. (2018). Dysregulation of valvular interstitial cell let-7c, miR-17, miR-20a, and miR-30d in naturally occurring canine myxomatous mitral valve disease. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0188617"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/j.prp.2005.02.001","article-title":"CD34+ fibrocytes in normal mitral valves and myxomatous mitral valve degeneration","volume":"201","author":"Barth","year":"2005","journal-title":"Pathol. Res. Pract."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"763","DOI":"10.2460\/ajvr.69.6.763","article-title":"Distribution of myofibroblasts, smooth muscle\u2013like cells, macrophages, and mast cells in mitral valve leaflets of dogs with myxomatous mitral valve disease","volume":"69","author":"Han","year":"2008","journal-title":"Am. J. Vet. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"347","DOI":"10.1038\/nrd3978","article-title":"Extracellular vesicles: Biology and emerging therapeutic opportunities","volume":"12","author":"Breakefield","year":"2013","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_18","first-page":"1372","article-title":"Roles of exosomes in cardioprotection","volume":"38","author":"Barile","year":"2016","journal-title":"Eur. Heart J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1350088","DOI":"10.1080\/20013078.2017.1350088","article-title":"Circulating exosome microRNA associated with heart failure secondary to myxomatous mitral valve disease in a naturally occurring canine model","volume":"6","author":"Yang","year":"2017","journal-title":"J. Extracell. Vesicles"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"eaav1741","DOI":"10.1126\/science.aav1741","article-title":"The biochemical basis of microRNA targeting efficacy","volume":"366","author":"McGeary","year":"2019","journal-title":"Science"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1038\/cr.2008.328","article-title":"Non-Smad pathways in TGF-beta signaling","volume":"19","author":"Zhang","year":"2009","journal-title":"Cell Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1161\/CIRCRESAHA.109.197517","article-title":"MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation","volume":"105","author":"Cheng","year":"2009","journal-title":"Circ. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1038\/nature08195","article-title":"miR-145 and miR-143 regulate smooth muscle cell fate and plasticity","volume":"460","author":"Cordes","year":"2009","journal-title":"Nature"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"28097","DOI":"10.1074\/jbc.M111.236950","article-title":"down-regulation of Kruppel-like factor-4 (KLF4) by microRNA-143\/145 is critical for modulation of vascular smooth muscle cell phenotype by transforming growth factor-beta and bone morphogenetic protein 4","volume":"286","author":"Chan","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1161\/CIRCGENETICS.110.958702","article-title":"miR-143 and miR-145: Molecular keys to switch the phenotype of vascular smooth muscle cells","volume":"4","author":"Rangrez","year":"2011","journal-title":"Circ. Cardiovasc. Genet."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1016\/j.yjmcc.2013.08.007","article-title":"Role of miR-145 in cardiac myofibroblast differentiation","volume":"66","author":"Wang","year":"2013","journal-title":"J. Mol. Cell. Cardiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2382","DOI":"10.1096\/fj.12-219493","article-title":"miR-145 regulates myofibroblast differentiation and lung fibrosis","volume":"27","author":"Yang","year":"2013","journal-title":"FASEB J."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Colombo, F., Norton, E.G., and Cocucci, E. (2022). Extracellular Vesicle exchange is favored by cell proximity. bioRxiv.","DOI":"10.1101\/2022.05.10.491399"},{"key":"ref_29","first-page":"E1","article-title":"Mir-30d Regulates Cardiac Remodeling by Intracellular and Paracrine Signaling","volume":"128","author":"Li","year":"2020","journal-title":"Circ. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1089\/scd.2018.0097","article-title":"Extracellular Vesicles from Wharton\u2019s Jelly Mesenchymal Stem Cells Suppress CD4 Expressing T Cells Through Transforming Growth Factor Beta and Adenosine Signaling in a Canine Model","volume":"28","author":"Crain","year":"2019","journal-title":"Stem Cells Dev."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5245","DOI":"10.1158\/0008-5472.CAN-04-0496","article-title":"Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets","volume":"64","author":"Andersen","year":"2004","journal-title":"Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"D155","DOI":"10.1093\/nar\/gky1141","article-title":"miRBase: From microRNA sequences to function","volume":"47","author":"Kozomara","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Love, M.I., Huber, W., and Anders, S. (2014). Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol., 15.","DOI":"10.1186\/s13059-014-0550-8"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH Image to ImageJ: 25 years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/3\/1468\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:49:05Z","timestamp":1760104145000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/3\/1468"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,1,25]]},"references-count":34,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2024,2]]}},"alternative-id":["ijms25031468"],"URL":"https:\/\/doi.org\/10.3390\/ijms25031468","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,1,25]]}}}