Abstract
Dystrophin deficiency due to genetic mutations causes cardiac abnormalities in Duchenne’s muscular dystrophy. Dystrophin is also shown to be downregulated in conventional failing hearts. Whether restoration of dystrophin expression possesses any therapeutic potential for conventional heart failure (HF) remains to be examined. HF mouse model was generated by transverse aortic constriction (TAC). In vivo activation of dystrophin transcription was achieved by tail-vein injection of adeno-associated virus 9 carrying CRISPR/dCas system for dystrophin. We found that activation of dystrophin expression in TAC mice significantly reduced the susceptibility to arrhythmia of TAC mice and the mortality rate. We further demonstrated that over-expression of dystrophin increased cardiac conduction of hearts in TAC mice by optical mapping evaluation. Activation of dystrophin expression also increased peak sodium current in isolated ventricular myocytes from hearts of TAC mice as recorded by the patch-clamp technique. Immunoblotting and immunofluorescence showed that increased dystrophin transcription restored the membrane distribution of Nav1.5 in the hearts of TAC mice. In summary, correction of dystrophin downregulation by the CRISPR-dCas9 system reduced the susceptibility to arrhythmia of conventional HF mice through restoring Nav1.5 membrane distribution. This study paved the way to develop a new therapeutic strategy for HF-related ventricular arrhythmia.
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Data availability
The data that support the findings of this study are available from the corresponding authors upon reasonable request.
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Funding
This work was supported by National Natural Science Foundation of China (82070344, 81870295 to ZP) and HMU Marshal Initiative Funding (HMUMIF-21017 to ZP).
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RZ, JL, and GX performed experiments, analyzed data, and prepared the manuscript. JY, DL, TT, XZ, and KG helped perform experiments and collect data. ZP designed the project, oversaw the experiments, and prepared the manuscript.
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Zhang, R., Liu, J., Xue, G. et al. Forced activation of dystrophin transcription by CRISPR/dCas9 reduced arrhythmia susceptibility via restoring membrane Nav1.5 distribution. Gene Ther 30, 142–149 (2023). https://doi.org/10.1038/s41434-022-00348-z
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DOI: https://doi.org/10.1038/s41434-022-00348-z