Pigs with δ-sarcoglycan deficiency exhibit traits of genetic cardiomyopathy

Hitomi Matsunari,Michiyo Honda,Masahito Watanabe,Satsuki Fukushima,Kouta Suzuki,Shigeru Miyagawa,Kazuaki Nakano,Kazuhiro Umeyama,Ayuko Uchikura,Kazutoshi Okamoto,Masaki Nagaya,Teruhiko Toyo-Oka,Yoshiki Sawa,Hiroshi Nagashima

doi:10.1038/s41374-020-0406-7

Abstract

Genetic cardiomyopathy is a group of intractable cardiovascular disorders involving heterogeneous genetic contribution. This heterogeneity has hindered the development of life-saving therapies for this serious disease. Genetic mutations in dystrophin and its associated glycoproteins cause cardiomuscular dysfunction. Large animal models incorporating these genetic defects are crucial for developing effective medical treatments, such as tissue regeneration and gene therapy. In the present study, we knocked out the δ-sarcoglycan (δ-SG) gene (SGCD) in domestic pig by using a combination of efficient de novo gene editing and somatic cell nuclear transfer. Loss of δ-SG expression in the SGCD knockout pigs caused a concomitant reduction in the levels of α-, β-, and γ-SG in the cardiac and skeletal sarcolemma, resulting in systolic dysfunction, myocardial tissue degeneration, and sudden death. These animals exhibited symptoms resembling human genetic cardiomyopathy and are thus promising for use in preclinical studies of next-generation therapies.

Highlights

Idiopathic cardiomyopathy is a serious cardiac disorder manifesting life-threatening regressive lesions of the heart muscle [1]
We knocked out SGCD in porcine cells by using transcription activator-like effector nucleases (TALENs) and produced genetically modified cloned animals by using somatic cell nuclear transfer (SCNT)
We found that homozygous KO of the SGCD in pigs induces a severe phenotype of genetic cardiomyopathy

Summary

Introduction

Idiopathic cardiomyopathy is a serious cardiac disorder manifesting life-threatening regressive lesions of the heart muscle [1]. The burden of this disease globally affects populations across all ages, sexes, and ethnic groups [2]. In hamsters of the TO-2 strain carrying the spontaneous biallelic mutation of the δ-SG gene (SGCD), DCM is highly prevalent [22, 23]. In contrast to the wealth of mutant laboratory rodents, the number of pig strains with targeted mutations is still inadequate [26] mainly due to technical limitations, the lack of a gene-knockout (KO) strategy involving the use of embryonic stem cells. We found that homozygous KO of the SGCD in pigs induces a severe phenotype of genetic cardiomyopathy

Methods

Results

Conclusion