Adeno-associated virus (AAV)-TBX18 does not generate biological pacemaker activity, unlike AAV-Hcn2
bioRxiv (Cold Spring Harbor Laboratory)
Abstract
Abstract Gene therapy-based biological pacemakers have been proposed as an alternative to their hardware-based counterparts. In this context, short-term ectopic expression of the T-box transcription factor 18 (TBX18) in the ventricle reportedly generated potent short-term pacemaker function in various animal models. Here, we investigated the effect of adeno-associated virus (AAV)-mediated long-term expression of TBX18, and compared the outcome to that of the pacemaker ion channel Hcn2. Our findings revealed that CMV-driven ectopic TBX18 expression in mouse hearts led to severe cardiac fibrosis. At lower, non-fibrogenic levels, TBX18 maintained its transcriptional function but failed to induce pacemaker phenotypes. TBX18-expressing cells showed suppressed expression of key working myocardial genes, but the pacemaker gene program was not induced. Electrophysiological studies showed abnormal automaticity in TBX18-expressing cells, combined with prolonged repolarization and various current changes. However, no hyperpolarization-activated funny current was detected. In a complete AV-block rat model, AAV-mediated Hcn2 expression induced robust ectopic pacemaker activity in the presence of isoproterenol, whereas TBX18 expression neither generated such activities, nor augmented Hcn2-mediated pacing. In conclusion, at functional non-fibrogenic levels, TBX18 is neither sufficient nor necessary to induce pacemaker activity. In contrast, Hcn2 generates reliable pacing, making it a more viable candidate for biological pacemaker development.
Authors 19
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Mathilde R. Rivaud Aachen Laboratory of Experimental Cardiology Department of Cardiology, Leiden University Medical Center
Leiden University Medical Center · Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Laboratory of Experimental Cardiology, Department of Cardiology, Leiden University Medical Center (LUMC), Albinusdreef 2, 2300 RC, Leiden, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
Laboratory of Experimental Cardiology, Department of Cardiology, Leiden University Medical Center (LUMC), Albinusdreef , 300 RC, Leiden, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 35, 1018 WB, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 5, 1018 WB, Amsterdam, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Affiliation as printed
Naason Science Inc, 123 Saengmyung-ro, Cheongju, Korea
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Revvity Gene Delivery (Germany)
Affiliation as printed
Naason Science Inc, 123 Saengmyung-ro, Cheongju, Korea
Revvity Gene Delivery GmbH, Am Haag 6, 82166 Graefelfing, Germany
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Witten/Herdecke University · Revvity Gene Delivery (Germany)
Affiliation as printed
Revvity Gene Delivery GmbH, Am Haag 6, 82166 Graefelfing, Germany
Virology and Microbiology, Center for Biomedical Education and Research, Department Human Medicine, Faculty of Health, University Witten/Herdecke, Stockumer Stre 10, 58455 Witten, Germany
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Witten/Herdecke University · Revvity Gene Delivery (Germany)
Affiliation as printed
Revvity Gene Delivery GmbH, Am Haag 6, 82166 Graefelfing, Germany
Virology and Microbiology, Center for Biomedical Education and Research, Department Human Medicine, Faculty of Health, University Witten/Herdecke, Stockumer Stre 10, 58455 Witten, Germany
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Witten/Herdecke University · Amsterdam University Medical Centers
Affiliation as printed
Virology and Microbiology, Center for Biomedical Education and Research, Department Human Medicine, Faculty of Health, University Witten/Herdecke, Stockumer Stre 10, 58455 Witten, Germany
Department of Anaesthesiology, Amsterdam University Medical Centers, De Boelelaan 111, 1081 HV, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 5, 1018 WB, Amsterdam, the Netherlands
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Witten/Herdecke University · Netherlands Heart Institute
Affiliation as printed
Virology and Microbiology, Center for Biomedical Education and Research, Department Human Medicine, Faculty of Health, University Witten/Herdecke, Stockumer Stre 10, 58455 Witten, Germany
Netherlands Heart Institute, Moreelsepark 1, 3511 EP, Utrecht, the Netherlands
PacingCure BV, Roetersstraat 5, 1018 WB, Amsterdam, the Netherlands
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Netherlands Heart Institute · Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Anaesthesiology, Amsterdam University Medical Centers, De Boelelaan 1117, 1081 HV, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 35, 1018 WB, Amsterdam, the Netherlands
Department of Clinical and Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Netherlands Heart Institute, Moreelsepark 1, 3511 EP, Utrecht, the Netherlands
PacingCure BV, Roetersstraat 5, 1018 WB, Amsterdam, the Netherlands
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Netherlands Heart Institute · Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Netherlands Heart Institute, Moreelsepark 1, 3511 EP, Utrecht, the Netherlands
PacingCure BV, Roetersstraat 35, 1018 WB, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Netherlands Heart Institute · Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Clinical and Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Netherlands Heart Institute, Moreelsepark 1, 3511 EP, Utrecht, the Netherlands
PacingCure BV, Roetersstraat 35, 1018 WB, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 5, 05 AZ, Amsterdam, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Clinical and Experimental Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
Department of Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 9, 15 AZ, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 5, 1018 WB, Amsterdam, the Netherlands
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Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
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Gerard J.J. Boink corresponding
Amsterdam University Medical Centers · Amsterdam Cardiovascular Sciences · University of Amsterdam
Affiliation as printed
Department of Cardiology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 9, 1105 AZ, Amsterdam, the Netherlands
Department of Medical Biology, Amsterdam Cardiovascular Sciences, Amsterdam University Medical Centers, University of Amsterdam, Meibergdreef 15, 1105 AZ, Amsterdam, the Netherlands
PacingCure BV, Roetersstraat 35, 1018 WB, Amsterdam, the Netherlands
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