Generation of a PKP2 heterozygous knockout pig model of arrhythmogenic cardiomyopathyOA
Arrhythmogenic cardiomyopathy(ACM)confers elevated risk of ventricular arrhythmias and sudden cardiac death,yet limitations in early lesion sampling and model development continue to hinder mechanistic and translational research.Clinical,pathological,and mutational profiles were examined in 24 individuals with ACM harboring PKP2 variants.Among these,a patient carrying the c.1132C>T mutation exhibited the earliest onset and presented both structural cardiac abnormalities and major adverse cardiovascular events.To facilitate disease modeling,the c.1147C>T variant—a previously reported pathogenic substitution located proximal to position c.1132 in PKP2—was selected to enhance the feasibility of generating a porcine model.The BE3 gene editing system was used to induce C>T mutation.Two single guide RNAs targeting the PKP2 gene were designed(sg RNA1 for c.1132C>T and sg RNA2 for c.1147C>T),yielding editing efficiencies of 42.9% and 25.9%,respectively.Sg RNA1 was used to generate PKP2^(+/-)porcine fetal fibroblasts.A total of 14 cloned piglets were produced,including 11 viable and three stillborn PKP2^(+/-)individuals.By 24 months of age,PKP2^(+/-)pigs developed premature ventricular contractions and right ventricular dilatation.Histological analysis revealed adipocyte infiltration within the right ventricular wall,and electron microscopy demonstrated reduced desmosomal length and electron density consistent with desmosomal dysfunction.Transcriptomic profiling showed high expression of genes associated with lipid catabolic processes.This study established the first PKP2^(+/-)porcine model of ACM using BE3-mediated base editing,providing a valuable platform for elucidating early pathogenic mechanisms and evaluating therapeutic interventions.
Hong-Hui Li;Yuan Chang;Tai-Yun Wei;Jian-Xiong Guo;De-Ling Jiao;Yi-Xuan Sheng;Hang Zhang;Han Mo;Pei-Yuan Li;Hai-Zhen Wang;Kai-Xiang Xu;Yu-Bo Qing;Heng Zhao;Bao-Yu Jia;Yan-Hua Su;Xiao Chen;Meng-Da Xu;Xiu-Meng Hua;Hao Cui;Hao Jia;Hong-Ye Zhao;Jiang-Ping Song;Hong-Jiang Wei
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Animal Science and Technology,Yunnan Agricultural University,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCollege of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,ChinaCardiomyopathy Research Group at Fuwai Hospital,State Key Laboratory of Cardiovascular Disease,National Center for Cardiovascular Diseases,Beijing 100037,China Beijing Key Laboratory of Preclinical Research and Evaluation for Cardiovascular Implant Materials,Animal Experimental Centre,Fuwai Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100037,China Fuwai Yunnan Hospital,Chinese Academy of Medical Sciences/Affiliated Cardiovascular Hospital of Kunming Medical University,Kunming,Yunnan 650102,ChinaState Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Yunnan Agricultural University,Kunming,Yunnan 650201,China Yunnan Key Laboratory of Porcine Gene Editing and Xenotransplantation,Kunming,Yunnan 650201,China College of Veterinary Medicine,Yunnan Agricultural University,Kunming,Yunnan 650201,China Kunming Institute of Zoology,Chinese Academy of Sciences,Kunming,Yunnan 650201,China
医药卫生
Arrhythmogenic cardiomyopathyPig modelBE3 gene-edit system
《Zoological Research》 2026 (1)
P.139-154,16
supported by the National Natural Science Foundation for Distinguished Young Scholars of China(82125004 to J.P.S.)Key Project of National Key R&D Program of the Ministry of Science and Technology of China(2023YFF0724700 to X.C.)Major Science and Technology Project of Yunnan Province(202102AA100054)Spring City Plan:High-level Talent Promotion and Training Project of Kunming(2022SCP001)。
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