Attenuation of Cardiomyocyte Hypertrophy via Depletion Myh7 using CASAAV
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Attenuation of Cardiomyocyte Hypertrophy via Depletion Myh7 using CASAAV Peng Yue1 · Shutao Xia2 · Gang Wu1 · Lei Liu1 · Kaiyu Zhou1 · Hongyu Liao1 · Jiawen Li1 · Xiaolan Zheng1 · Yuxuan Guo3 · Yimin Hua1 · Donghui Zhang2 · Yifei Li1 Received: 10 June 2020 / Accepted: 16 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Myh7 is a classic biomarker for cardiac remodeling and a potential target to attenuate cardiomyocyte (CM) hypertrophy. This study aimed to identify the dominant function of Myh7 after birth and determine whether its removal would affect CM maturation or contribute to reversal of pathological hypertrophy phenotypes. The CASAAV (CRISPR/Cas9-AAV9based somatic mutagenesis) technique was used to deplete Myh6 and Myh7, and an AAV dosage of 5 × 109 vg/g was used to generate a mosaic CM depletion model to explore the function of Myh7 in adulthood. CM hypertrophy was induced by transverse aortic constriction (TAC) in Rosa26Cas9-P2A-GFP mice at postnatal day 28 (PND28). Heart function was measured by echocardiography. Isolated CMs and in situ imaging were used to analyze the structure and morphology of CM. We discovered that CASAAV successfully silenced Myh6 and Myh7 in CMs, and early depletion of Myh7 led to mild adulthood lethality. However, the Myh7 PND28-knockout mice had normal heart phenotype and function, with normal cellular size and normal organization of sarcomeres and T-tubules. The TAC mice also received AAV-Myh7-Cre to produce Myh7-knockout CMs, which were also of normal size, and echocardiography demonstrated a reversal of cardiac hypertrophy. In conclusion, Myh7 has a role during the maturation period but rarely functions in adulthood. Thus, the therapeutic time should exceed the period of maturation. These results confirm Myh7 as a potential therapeutic target and indicate that its inhibition could help reverse CM hypertrophy. Keywords Myh7 · Myh6 · CASAAV · Cardiomyocyte hypertrophy
Introduction
Communicated by Shazina Saeed. Peng Yue and Shutao Xia contributed equally to this work. * Donghui Zhang [email protected] * Yifei Li [email protected] 1
Key Laboratory of Birth Defects and Related Diseases of Women and Children of MOE, Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu 610041, Sichuan, China
2
State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Science, Hubei University, Wuhan 430062, Hubei, China
3
Department of Cardiology, Boston Children’s Hospital, Boston, MA 02115, USA
Cardiomyocyte (CM) hypertrophy is a common pathophysiological process occurring in several cardiovascular diseases [1]. Cardiac hypertrophy is the fundamental organic change that precedes functional changes in the heart, and its main manifestation is pathological remodeling [2]. During pathological remodeling in heart disease, multiple genes related to heart function show re-coding. These include genes encoding contractile phenotype proteins, which are directly related to cardiac
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