Deep phenotyping of testosterone-prompted fibrosis in arrhythmogenic right ventricular cardiomyopathy using iPSC-derived engineered cardiac spheroids  

睾酮加重遗传性心肌病心脏类器官球体纤维化

作　　者：Hongyi Cheng Xinrui Wang Sichong Qian Yike Zhang Jincheng Jiao Bingyu Zheng Yue Zhu Hua Xu Jia Song Feng Zhang Xiaohong Jiang Chang Cui Minglong Chen 程弘毅;王心睿;钱思翀;张易轲;焦锦程;郑冰玉;竺悦;徐华;宋佳;张峰;姜晓宏;崔畅;陈明龙(南京医科大学第一附属医院心内科,南京市210029;南京医科大学姑苏学院,苏州市215002;福建医科大学妇儿临床医学院,福州市350122;福建省妇幼保健院福建省妇儿重大疾病研究重点实验室,福州市350001;北京安贞医院心脏外科,北京市100029;东南大学生物科学与医学工程学院数字医学工程全国重点实验室,南京市210096;贝勒医学院,美国休斯顿770308;南京医科大学心血管病转化医学协同创新中心江苏省心血管病靶向干预研究重点实验室,南京市210029)

机构地区：[1]Department of Cardiology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China [2]Gusu School,Nanjing Medical University,Suzhou 215002,China [3]Medical Research Center,Fujian Maternity and Child Health Hospital,College of Clinical Medicine for Obstetrics&Gynecology and Pediatrics,Fujian Medical University,Fuzhou 350122,China [4]Fujian Key Laboratory of Women and Children’s Critical Diseases Research,Fujian Maternity and Child Health Hospital,Fuzhou 350001,China [5]Department of Cardiac Surgery,Anzhen Hospital,Beijing 100029,China [6]State Key Laboratory of Digital Medical Engineering,School of Biological Science and Medical Engineering,Southeast University,Nanjing 210096,China [7]Department of Medicine,Section of Cardiovascular Research,Baylor College of Medicine,Houston TX 77030,USA [8]Key Laboratory of Targeted Intervention of Cardiovascular Disease,Collaborative Innovation Center for Cardiovascular Disease Translational Medicine,Nanjing Medical University,Nanjing 210029,China

出　　处：《Bio-Design and Manufacturing》2025年第1期20-35,I0002-I0012,共27页生物设计与制造(英文)

基　　金：supported by the National Natural Science Foundation of China(Nos.82370322 to CC,82200352 to FZ,82300352 to YZ,22275034 to HX,and 82070343 to MLC);the Natural Science Foundation of Jiangsu Province of China(Nos.BK20220710 to FZ and BK20230733 to YZ);Postgraduate Research&Practice Innovation Program of Jiangsu Province(No.JX13414086 to HYC).

摘　　要：Arrhythmogenic right ventricular cardiomyopathy(ARVC)is a progressive disease characterized by adipose and fibrous replacement of the myocardium.While elevated testosterone levels have been implicated in the pathological process of ARVC,its exact contribution to cardiac fibrosis in ARVC remains unclear.In this study,we analyzed the potential contribution of gender-based differences on the distribution of the low-voltage area in an ARVC cohort undergoing an electrophysiological study,which was indicated by feature selection.Additionally,we established engineered cardiac spheroid models in vitro using patient-specific induced pluripotent stem cell(iPSC)-derived cardiomyocytes(iPSC-CMs)and iPSC-derived cardiac fibroblasts(icFBs).We elucidated the pathogenicity of abnormal splicing in the plakophilin-2(PKP2)gene caused by an intronic mutation.Additionally,pathogenic validation of the desmoglein-2(DSG2)point mutation further confirms the reliability of the models.Moreover,testosterone exacerbated the DNA damage in the mutated cardiomyocytes and further activated myofibroblasts in a chain reaction.In conclusion,we designed and constructed an in vitro three-dimensionally-engineered cardiac spheroid model of ARVC based on clinical findings and provided direct evidence of the fibrotic role of testosterone in ARVC.致心律失常性右室心肌病(ARVC)是一种遗传性心肌病,进行性脂肪纤维化是其独特特征。升高的睾酮水平可能会促进ARVC的纤维化病理过程。然而,因为缺乏良好的遗传性心肌病模型,睾酮对ARVC心脏纤维化的具体作用仍不明确。此研究首先在接受了临床电生理标测的ARVC患者中明确了纤维化低电压区分布的性别差异,接着使用患者特异性诱导多能干细胞来源的心肌细胞和心脏成纤维细胞构建了三维心脏类器官球体模型,进而在该模型上使用睾酮进行了药物试验。与动物模型比,体外的心脏类器官球体平台具有更好的代表性,观察疾病表型更加直观便捷。同时基于诱导多能干细胞分化的多种细胞类型构建的心脏类器官是更加生理化的多细胞共培养模型,可用于探究细胞间相互作用的机制,明确致病的靶细胞和靶分子,有利于遗传性心肌病的精准干预。总之,该研究建立了一种新的ARVC体外三维模型,并提供了直接证据表明睾酮在ARVC中起到了促纤维化的作用。

关 键 词：Arrhythmogenic right ventricular cardiomyopathy(ARVC) Gender difference Cardiac spheroids Testoste-rone FIBROSIS 

分 类 号：R542.2[医药卫生—心血管疾病]