Cyclin A2 Induces Human Adult Cardiomyocyte Cytokinesis and Elicits Cardiomyocyte Reprogramming and Dedifferentiation

Cyclin A2 Induces Human Adult Cardiomyocyte Cytokinesis and Elicits Cardiomyocyte Reprogramming and Dedifferentiation | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Cyclin A2 Induces Human Adult Cardiomyocyte Cytokinesis and Elicits Cardiomyocyte Reprogramming and Dedifferentiation Esmaa Bouhamida, Sangeetha Vadakke-Madathil, Prabhu Mathiyalagan, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6597490/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 03 Nov, 2025 Read the published version in npj Regenerative Medicine → Version 1 posted 9 You are reading this latest preprint version Abstract Cyclin A2 (CCNA2), a master cell cycle regulator, is silenced in postnatal mammalian cardiomyocytes. We have previously demonstrated its ability to promote cardiac repair in small and large animals when delivered to the heart via a viral vector. However, the effect of CCNA2 gene delivery on cytokinesis in isolated cardiomyocytes from adult human hearts has not been explored. We designed a human gene therapy vector featuring a replication-deficient adenovirus encoding human CCNA2 driven by the cardiac Troponin T promoter to enable the expression of CCNA2 in freshly isolated human cardiomyocytes. Time-lapse live imaging of adult human primary cardiomyocytes from a 21-year-old male, a 41-year-old female, and a 55-year-old male demonstrated the induction of complete cytokinesis in human adult cardiomyocytes with preservation of sarcomere integrity in the resulting daughter cells with active calcium mobilization in redifferentiated cardiomyocytes. To elucidate the transcriptional mechanisms underlying this response, we conducted single-nucleus transcriptomics analysis of hearts isolated from adult transgenic mice that constitutively express CCNA2 in cardiomyocytes (CCNA2-Tg) and non- transgenic mice (nTg). This revealed a cardiomyocyte subpopulation enriched with cytokinesis, proliferative, and reprogramming genes in hearts obtained from CCNA2-Tg mice as compared to41nTg mice. Ultra-deep bulk RNA sequencing of human adult and fetal hearts identified key reprogramming genes relevant to understanding the mechanisms of CCNA2-induced effects observed in our experimental models. These findings provide a promising path for the clinical development of CCNA2-based cardiac regenerative therapy. Health sciences/Medical research/Translational research Biological sciences/Biotechnology Biological sciences/Cell biology Biological sciences/Genetics Biological sciences/Stem cells Health sciences/Medical research Full Text Additional Declarations Competing interest reported. H.W.C. is the inventor of multiple patents regarding cyclin A2-mediated cardiac repair and caudal-related homeobox 2 cells for cardiac repair. She is in the midst of launching a new biotechnology company to advance such therapies to clinical development. Other authors declare no financial competing interests. Supplementary Files Therealtimeliveimagingmovieofhumanadultcardiomyocytes55yearoldmaleisdescribedinFigure1.avi Movie S1. The real-time, live-imaging movie of human adult cardiomyocytes (55-year-old male) as described in Figure 1. Therealtimeliveimagingmovieofhumanadultcardiomyocytes41yearoldFemaleisdescribedinFigure1.mov Movie S2. Real-time, live-imaging movie of human adult cardiomyocytes (41-year-old female) as described in Figure 1. Cite Share Download PDF Status: Published Journal Publication published 03 Nov, 2025 Read the published version in npj Regenerative Medicine → Version 1 posted Editorial decision: Revision requested 05 Jul, 2025 Reviews received at journal 27 Jun, 2025 Reviews received at journal 20 Jun, 2025 Reviewers agreed at journal 01 Jun, 2025 Reviewers agreed at journal 23 May, 2025 Reviewers invited by journal 13 May, 2025 Editor assigned by journal 13 May, 2025 Submission checks completed at journal 06 May, 2025 First submitted to journal 05 May, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6597490","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":456767706,"identity":"252e1c94-01fe-4d81-8730-c440f8ddccf6","order_by":0,"name":"Esmaa Bouhamida","email":"","orcid":"","institution":"Icahn School of Medicine at Mount Sinai","correspondingAuthor":false,"prefix":"","firstName":"Esmaa","middleName":"","lastName":"Bouhamida","suffix":""},{"id":456767707,"identity":"79a47548-131e-4d50-a1a6-d57097c26ec9","order_by":1,"name":"Sangeetha Vadakke-Madathil","email":"","orcid":"","institution":"Icahn School of Medicine at Mount Sinai","correspondingAuthor":false,"prefix":"","firstName":"Sangeetha","middleName":"","lastName":"Vadakke-Madathil","suffix":""},{"id":456767708,"identity":"1f25a4a3-4080-48e7-a6ff-9ad02f332b30","order_by":2,"name":"Prabhu Mathiyalagan","email":"","orcid":"","institution":"Benthos Prime Central, TX, USA","correspondingAuthor":false,"prefix":"","firstName":"Prabhu","middleName":"","lastName":"Mathiyalagan","suffix":""},{"id":456767709,"identity":"85e5d68d-a616-431e-b044-6e569ea2373b","order_by":3,"name":"Amaresh K. 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We have previously demonstrated its ability to promote cardiac repair in small and large animals when delivered to the heart via a viral vector. However, the effect of CCNA2 gene delivery on cytokinesis in isolated cardiomyocytes from adult human hearts has not been explored. We designed a human gene therapy vector featuring a replication-deficient adenovirus encoding human CCNA2 driven by the cardiac Troponin T promoter to enable the expression of CCNA2 in freshly isolated human cardiomyocytes. Time-lapse live imaging of adult human primary cardiomyocytes from a 21-year-old male, a 41-year-old female, and a 55-year-old male demonstrated the induction of complete cytokinesis in human adult cardiomyocytes with preservation of sarcomere integrity in the resulting daughter cells with active calcium mobilization in redifferentiated cardiomyocytes. 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