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The drug-elicitable alternative splicing module for tunable vector expression in the heart

  • 1Institute of Cardiovascular Sciences, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
  • 2State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, China.
  • 3Department of Biomedical Informatics, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
  • 4Vituner Therapeutics, Nantong, China.
  • 5Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China.
  • 6State Key Laboratory of Natural and Biomimetic Drugs, Beijing Key Laboratory of Molecular Pharmaceutics, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing, China.
  • 7Ministry of Education Key Laboratory of Birth Defects and Related Diseases of Women and Children, Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, China.
  • 8Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing, China.
  • 9Department of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital, Beijing, China.
  • 10Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, China.
  • 11Department of Cardiology, Boston Children's Hospital, Boston, MA, USA.
  • 12Zhuhai Hengqin SBCVC Xinchuang Equity Investment Management Enterprise (Limited Partnership), Zhuhai, China.
  • 13Institute of Cardiovascular Sciences, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China. donged@bjmu.edu.cn.
  • 14State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, China. donged@bjmu.edu.cn.
  • 15Department of Cardiology and Institute of Vascular Medicine, Peking University Third Hospital, Beijing, China. donged@bjmu.edu.cn.
  • 16Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, China. donged@bjmu.edu.cn.
  • 17Research Center for Cardiopulmonary Rehabilitation, University of Health and Rehabilitation Sciences Qingdao Hospital (Qingdao Municipal Hospital), School of Health and Life Sciences, University of Health and Rehabilitation Sciences, Qingdao, China. donged@bjmu.edu.cn.
  • 18Institute of Cardiovascular Sciences, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, China. guo@bjmu.edu.cn.
  • 19State Key Laboratory of Vascular Homeostasis and Remodeling, Peking University, Beijing, China. guo@bjmu.edu.cn.
  • 20Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing, China. guo@bjmu.edu.cn.
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June 13, 2025

Abstract

Adeno-associated viruses (AAVs) are commonly used for gene therapy, but a clinically relevant method to fine-tune AAV expression is lacking, restricting their therapeutic efficacy and safety. Here we develop the drug-elicitable alternative splicing module (DreAM), which is responsive to risdiplam, a Food and Drug Administration-approved alternative splicing modulator. Risdiplam activated DreAM-regulated AAV expression in a dose-dependent manner with a 2,000-fold inducible change, depending on the dose of risdiplam and the organ of interest. With a temporal resolution of 2 days, DreAM could transiently, reversibly and repeatedly activate AAV expression according to the frequency and duration of risdiplam administration. In this proof-of-concept study, we incorporated DreAM into the cardiomyocyte-specific, liver-detargeted AAV9-Tnnt2-miR122TS vector to transiently activate the cardiomyocyte regeneration factor YAP5SA. A dedifferentiation-proliferation-redifferentiation cycle was established in adult cardiomyocytes, improving cardiac regeneration after myocardial infarction while limiting animal death, AAV9-Tnnt2 expression in the liver and hepatic tumorigenesis. Therefore, DreAM may enhance the efficacy, safety and scope of gene therapy.