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Homology-directed repair of an MYBPC3 gene mutation in a rat model of hypertrophic cardiomyopathy

Gene Therapy volume 30, pages 520–527 (2023)Cite this article

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Abstract

Variants in myosin-binding protein C3 (MYBPC3) gene are a main cause of hypertrophic cardiomyopathy (HCM), accounting for 30% to 40% of the total number of HCM mutations. Gene editing represents a potential permanent cure for HCM. The aim of this study was to investigate whether genome editing of MYBPC3 using the CRISPR/Cas9 system in vivo could rescue the phenotype of rats with HCM. We generated a rat model of HCM (“1098hom”) that carried an Mybpc3 premature termination codon mutation (p.W1098x) discovered in a human HCM pedigree. On postnatal day 3, the CRISPR/Cas9 system was introduced into rat pups by a single dose of AAV9 particles to correct the variant using homology-directed repair (HDR). Analysis was performed 6 months after AAV9 injection. The 1098hom rats didn’t express MYBPC3 protein and developed an HCM phenotype with increased ventricular wall thickness and diminished cardiac function. Importantly, CRISPR HDR genome editing corrected 3.56% of total mutations, restored MYBPC3 protein expression by 2.12%, and normalized the HCM phenotype of 1098hom rats. Our work demonstrates that the HDR strategy is a promising approach for treating HCM associated with MYBPC3 mutation, and that CRISPR technology has great potential for treating hereditary heart diseases.

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Fig. 1: Generation of the 1098hom rat model of HCM.
Fig. 2: Designation of CRISPR/Cas9 HDR strategy to correct the p.W1098x mutation.
Fig. 3: HDR treatment alleviated cardiac hypertrophy in 1098hom rats.
Fig. 4: HDR treatment improved cardiac function in 1098hom rats.
Fig. 5: Evaluation of HDR editing efficiency.
Fig. 6: HDR treatment partially restored MYBPC3 protein expression.

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Data availability

The datasets analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We acknowledge and appreciate our colleagues for their valuable suggestions and technical assistance for this study.

Funding

This work was supported by National Natural Science Foundation of China [No. 82100401, 82070354, 81470519, 81630010] and Huazhong University of Science and Technology Academic Frontier Youth Team (No. 2019QYTD08).

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Author notes

  1. These authors contributed equally: Jiali Nie; Yu Han.

Authors and Affiliations

  1. Division of Cardiology, Department of Internal Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Jiali Nie, Yu Han, Weijian Hang, Hongyang Shu, Zheng Wen, Li Ni & Dao Wen Wang

  2. Hubei Key Laboratory of Genetics and Molecular Mechanisms of Cardiological Disorders, Wuhan, 430030, China

    Jiali Nie, Yu Han, Weijian Hang, Hongyang Shu, Zheng Wen, Li Ni & Dao Wen Wang

  3. Department of Cardiology, Xiamen Cardiovascular Hospital, Xiamen University, Xiamen, 361000, China

    Zhiyuan Jin

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Contributions

DWW, LN and JN designed the study. JN and YH conducted the experiments, analyzed data and completed the manuscript. ZJ, WH and HS conducted the animal experiments. ZW performed the echocardiography examination of the animals.

Corresponding authors

Correspondence to Li Ni or Dao Wen Wang.

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Competing interests

The authors declare no competing interests.

Ethics approval

The study was approved by the Ethics Review Board of Tongji Hospital and Tongji Medical College. It complied with the principles of the Declaration of Helsinki. Written informed consent was obtained from individual subjects of the HCM pedigree in this study. All animal experiments complied with the “Guide for the Care and Use of Laboratory Animals” published by the United States National Institutes of Health (NIH Publication No. 85-23, revised 1996). This study was approved by the Institutional Animal Research Committee of Tongji Medical College.

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Nie, J., Han, Y., Jin, Z. et al. Homology-directed repair of an MYBPC3 gene mutation in a rat model of hypertrophic cardiomyopathy. Gene Ther 30, 520–527 (2023). https://doi.org/10.1038/s41434-023-00384-3

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