{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,8,11]],"date-time":"2026-08-11T00:29:31Z","timestamp":1786408171225,"version":"build-2736575974"},"posted":{"date-parts":[[2026,8,7]]},"group-title":"Cell Biology","reference-count":40,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T00:00:00Z","timestamp":1786060800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,8,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Duchenne muscular dystrophy (DMD) cardiomyopathy is characterized by progressive microtubule remodeling, connexin-43 (Cx43) dysregulation, and ventricular arrhythmias. We previously demonstrated phospho-mimic knock-in of \u03b2III-tubulin S172E preserves microtubule organization and attenuates cardiac pathology in mdx mice. However, whether these protective effects can be reproduced using a clinically relevant gene-delivery strategy remains unknown.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods and Results<\/jats:title>\n                  <jats:p>We generated a cardiomyocyte-specific adeno-associated virus serotype 9 (AAV9) vector expressing phospho-mimic \u03b2III-tubulin (Tubb3-S172E) under the cardiac troponin T promoter and delivered it to 4-5-month-old wild-type and mdx mice. Cardiac Tubb3-S172E expression was confirmed by quantitative qPCR and immunoblotting. In mdx mice, AAV9-mediated Tubb3-S172E expression significantly reduced mononuclear inflammatory infiltration, restored Cx43 localization at intercalated discs, and attenuated isoproterenol-induced arrhythmia susceptibility. In contrast, cardiac fibrosis, Nav1.5 protein expression, and peak sodium current density were not significantly improved. Overexpression of wild-type \u03b2III-tubulin in healthy hearts increased Cx43 lateralization and arrhythmia susceptibility, indicating that \u03b2III-tubulin phosphorylation state rather than protein abundance determines its protective function.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Cardiomyocyte-targeted delivery of phospho-mimic \u03b2III-tubulin partially recapitulates the protective effects observed in the genetic S172E knock-in model. These findings identify \u03b2III-tubulin Ser172 phosphorylation as a critical regulator of microtubule-dependent electrical remodeling and support therapeutic modulation of this pathway in Duchenne muscular dystrophy cardiomyopathy.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Research Perspective<\/jats:title>\n                  <jats:list list-type=\"bullet\">\n                    <jats:list-item>\n                      <jats:p>Cardiomyocyte-targeted AAV9 delivery of phospho-mimic \u00e2III-tubulin improves Cx43 organization, inflammatory remodeling, and arrhythmia susceptibility in dystrophic hearts, demonstrating that therapeutic modulation of \u03b2III-tubulin Ser172 phosphorylation partially recapitulates the protective effects observed in the genetic S172E model.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>The dissociation between improved electrical remodeling and persistent Nav1.5 and fibrotic abnormalities suggests that \u03b2III-tubulin Ser172 phosphorylation selectively regulates specific microtubule-dependent pathological pathways in dystrophic cardiomyopathy.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>Future studies should define the molecular mechanisms linking \u03b2III-tubulin Ser172 phosphorylation to cardiomyocyte-immune cell communication and determine how this pathway coordinates electrical and inflammatory remodeling in dystrophic hearts.<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.64898\/2026.08.04.742904","type":"posted-content","created":{"date-parts":[[2026,8,7]],"date-time":"2026-08-07T16:50:19Z","timestamp":1786121419000},"source":"Crossref","is-referenced-by-count":0,"title":["AAV9-mediated \u03b2III-tubulin Ser172 phospho-mimic expression improves arrhythmic and inflammatory remodeling in dystrophic cardiomyopathy"],"prefix":"10.64898","author":[{"given":"Delong","family":"Zhou","sequence":"first","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasisht","family":"Yegneshwaran","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nehal Kamal","family":"Ali","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geovanni","family":"Geukgeuzian","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elam","family":"Mesa","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2600-2269","authenticated-orcid":false,"given":"Lai-Hua","family":"Xie","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5369-4446","authenticated-orcid":false,"given":"Diego","family":"Fraidenraich","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Molecular Medicine, New Jersey Medical School, Rutgers Biomedical and Health Sciences, Newark, NJ, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"issue":"5","key":"2026081016400794000_2026.08.04.742904v1.1","first-page":"00176","article-title":"Cardiomyopathy as cause of death in Duchenne muscular dystrophy: a longitudinal observational study","volume":"9","year":"2023","journal-title":"ERJ Open Res"},{"issue":"6","key":"2026081016400794000_2026.08.04.742904v1.2","first-page":"608","article-title":"Cardiovascular Disease in Duchenne Muscular Dystrophy","volume":"7","year":"2022","journal-title":"JACC: Basic to Translational 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