{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T03:16:02Z","timestamp":1775618162239,"version":"3.50.1"},"reference-count":82,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2022,8,17]],"date-time":"2022-08-17T00:00:00Z","timestamp":1660694400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council of Australia Project","doi-asserted-by":"publisher","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council of Australia Project","doi-asserted-by":"publisher","award":["100463"],"award-info":[{"award-number":["100463"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001030","name":"National Heart Foundation of Australia (JJHC)","doi-asserted-by":"publisher","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}],"id":[{"id":"10.13039\/501100001030","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001030","name":"National Heart Foundation of Australia (JJHC)","doi-asserted-by":"publisher","award":["100463"],"award-info":[{"award-number":["100463"]}],"id":[{"id":"10.13039\/501100001030","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Sydney Medical School Foundation Fellowship (JJHC)","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}]},{"name":"Sydney Medical School Foundation Fellowship (JJHC)","award":["100463"],"award-info":[{"award-number":["100463"]}]},{"name":"MyWestmead Early Career Research Scholarship","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}]},{"name":"MyWestmead Early Career Research Scholarship","award":["100463"],"award-info":[{"award-number":["100463"]}]},{"name":"Arab Bank Postgraduate Research Scholarship","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}]},{"name":"Arab Bank Postgraduate Research Scholarship","award":["100463"],"award-info":[{"award-number":["100463"]}]},{"name":"Westmead Medical Research Foundation","award":["APP1128864"],"award-info":[{"award-number":["APP1128864"]}]},{"name":"Westmead Medical Research Foundation","award":["100463"],"award-info":[{"award-number":["100463"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Sinoatrial node dysfunction can manifest as bradycardia, leading to symptoms of syncope and sudden cardiac death. Electronic pacemakers are the current standard of care but are limited due to a lack of biological chronotropic control, cost of revision surgeries, and risk of lead- and device-related complications. We therefore aimed to develop a biological alternative to electronic devices by using a clinically relevant gene therapy vector to demonstrate conversion of cardiomyocytes into sinoatrial node-like cells in an in vitro context. Neonatal rat ventricular myocytes were transduced with recombinant adeno-associated virus vector 6 encoding either hTBX18 or green fluorescent protein and maintained for 3 weeks. At the endpoint, qPCR, Western blot analysis and immunocytochemistry were used to assess for reprogramming into pacemaker cells. Cell morphology and Arclight action potentials were imaged via confocal microscopy. Compared to GFP, hTBX18-transduced cells showed that hTBX18, HCN4 and Cx45 were upregulated. Cx43 was significantly downregulated, while sarcomeric \u03b1-actinin remained unchanged. Cardiomyocytes transduced with hTBX18 acquired the tapering morphology of native pacemaker cells, as compared to the block-like, striated appearance of ventricular cardiomyocytes. Analysis of the action potentials showed phase 4 depolarization and a significant decrease in the APD50 of the hTBX18-transduced cells. We have demonstrated that rAAV-hTBX18 gene transfer to ventricular myocytes results in morphological, molecular, physiological, and functional changes, recapitulating the pacemaker phenotype in an in vitro setting. The generation of these induced pacemaker-like cells using a clinically relevant vector opens new prospects for biological pacemaker development.<\/jats:p>","DOI":"10.3390\/ijms23169230","type":"journal-article","created":{"date-parts":[[2022,8,17]],"date-time":"2022-08-17T21:23:56Z","timestamp":1660771436000},"page":"9230","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Recombinant Adeno-Associated Viral Vector-Mediated Gene Transfer of hTBX18 Generates Pacemaker Cells from Ventricular Cardiomyocytes"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0656-2816","authenticated-orcid":false,"given":"Melad","family":"Farraha","sequence":"first","affiliation":[{"name":"Sydney Medical School, the University of Sydney, Sydney 2006, Australia"},{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9437-3761","authenticated-orcid":false,"given":"Renuka","family":"Rao","sequence":"additional","affiliation":[{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sindhu","family":"Igoor","sequence":"additional","affiliation":[{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thi Y. L.","family":"Le","sequence":"additional","affiliation":[{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael A.","family":"Barry","sequence":"additional","affiliation":[{"name":"Department of Cardiology, Westmead Hospital, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Davey","sequence":"additional","affiliation":[{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"},{"name":"School of Physics, the University of Sydney, Sydney 2006, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cindy","family":"Kok","sequence":"additional","affiliation":[{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James J.H.","family":"Chong","sequence":"additional","affiliation":[{"name":"Sydney Medical School, the University of Sydney, Sydney 2006, Australia"},{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"},{"name":"Department of Cardiology, Westmead Hospital, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Eddy","family":"Kizana","sequence":"additional","affiliation":[{"name":"Sydney Medical School, the University of Sydney, Sydney 2006, Australia"},{"name":"Centre for Heart Research, the Westmead Institute for Medical Research, Sydney 2145, Australia"},{"name":"Department of Cardiology, Westmead Hospital, Sydney 2145, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1038\/nbt.2465","article-title":"Transcription factor-driven conversion of quiescent cardiomyocytes to pacemaker cells","volume":"31","author":"Kapoor","year":"2013","journal-title":"Nat. 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