Phenotypic recapitulation and correction of desmoglein-2-deficient cardiomyopathy using human-induced pluripotent stem cell-derived cardiomyocytes.
Phenotypic recapitulation and correction of desmoglein-2-deficient cardiomyopathy using human-induced pluripotent stem cell-derived cardiomyocytes.
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DOI：
10.1093/hmg/ddab127
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发表时间：
2021-07-09
影响因子：
3.5
通讯作者：
Sakata Y
中科院分区：
文献类型：
--
作者：
Shiba M;Higo S;Kondo T;Li J;Liu L;Ikeda Y;Kohama Y;Kameda S;Tabata T;Inoue H;Nakamura S;Takeda M;Ito E;Takashima S;Miyagawa S;Sawa Y;Hikoso S;Sakata Y
桥粒蛋白-2由DSG2编码，是维持包括心脏在内的组织结构完整性的桥粒蛋白之一。DSG2基因突变主要以常染色体显性方式引起致心律失常的心肌病。在这里，我们鉴定了DSG2(c.C355T，p.R119X)的纯合子停止-增益突变，该突变导致严重双室心衰患者的桥粒芯糖蛋白-2完全缺乏。组织学分析显示心肌中桥粒蛋白异常沉积，破坏间盘结构。从患者(R119X-IPSC)中获得诱导多能干细胞(IPSC)，并通过同源定向修复(HDR-IPSC)将突变的DSG2基因位点杂合校正为正常等位基因。两种同基因IPSC均分化为心肌细胞[诱导多能干细胞来源的心肌细胞(IPSC-CMS)]。多电极阵列分析在R119X-IPSC-CMS中检测到异常激发，而在HDR-IPSC-CMS中未发现异常激发。三维自组织组织环的微力测试显示，R119X-IPSC-CMS的组织脆性和组织环的最大力较弱。值得注意的是，这些表型在HDR-IPSC-CMS中显著恢复。R119X-iPSC-CMS的心肌纤维结构严重异常，电子显微镜分析证实这些细胞内桥粒断裂。出乎意料的是，R119X-IPSC-CMS中桥粒蛋白-2的缺失导致桥粒蛋白-2的表达减少，但没有其他桥粒蛋白的表达。腺相关病毒介导的DSG2替换显著恢复了R119X-IPSC-CMS产生的SOTRs的收缩力量。我们的发现证实了在临床诊断的扩张型心肌病中存在桥粒芯糖蛋白-2缺乏的心肌病。IPSC-CMS对疾病表型的概括和纠正为精确医学的发展提供了证据，并为这种心肌病的基因替代治疗提供了概念验证。
Desmoglein-2, encoded by DSG2, is one of the desmosome proteins that maintain the structural integrity of tissues, including heart. Genetic mutations in DSG2 cause arrhythmogenic cardiomyopathy, mainly in an autosomal dominant manner. Here, we identified a homozygous stop-gain mutations in DSG2 (c.C355T, p.R119X) that led to complete desmoglein-2 deficiency in a patient with severe biventricular heart failure. Histological analysis revealed abnormal deposition of desmosome proteins, disrupted intercalated disk structures in the myocardium. Induced pluripotent stem cells (iPSCs) were generated from the patient (R119X-iPSC), and the mutated DSG2 gene locus was heterozygously corrected to a normal allele via homology-directed repair (HDR-iPSC). Both isogenic iPSCs were differentiated into cardiomyocytes [induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs)]. Multielectrode array analysis detected abnormal excitation in R119X-iPSC-CMs but not in HDR-iPSC-CMs. Micro-force testing of three-dimensional self-organized tissue rings (SOTRs) revealed tissue fragility and a weak maximum force in SOTRs from R119X-iPSC-CMs. Notably, these phenotypes were significantly recovered in HDR-iPSC-CMs. Myocardial fiber structures in R119X-iPSC-CMs were severely aberrant, and electron microscopic analysis confirmed that desmosomes were disrupted in these cells. Unexpectedly, the absence of desmoglein-2 in R119X-iPSC-CMs led to decreased expression of desmocollin-2 but no other desmosome proteins. Adeno-associated virus-mediated replacement of DSG2 significantly recovered the contraction force in SOTRs generated from R119X-iPSC-CMs. Our findings confirm the presence of a desmoglein-2-deficient cardiomyopathy among clinically diagnosed dilated cardiomyopathies. Recapitulation and correction of the disease phenotype using iPSC-CMs provide evidence to support the development of precision medicine and the proof of concept for gene replacement therapy for this cardiomyopathy.
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影响因子：
4.6
作者：
Kohama Y;Higo S;Masumura Y;Shiba M;Kondo T;Ishizu T;Higo T;Nakamura S;Kameda S;Tabata T;Inoue H;Motooka D;Okuzaki D;Takashima S;Miyagawa S;Sawa Y;Hikoso S;Sakata Y
通讯作者：
Sakata Y
影响因子：
9.5
作者：
Krusche CA;Holthöfer B;Hofe V;van de Sandt AM;Eshkind L;Bockamp E;Merx MW;Kant S;Windoffer R;Leube RE
通讯作者：
Leube RE
影响因子：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
8
作者：
Chelko, Stephen P.;Asimaki, Angeliki;Judge, Daniel P.
通讯作者：
Judge, Daniel P.
影响因子：
39.3
作者：
Basso, Cristina;Czarnowska, Elzbieta;Rampazzo, Alessandra
通讯作者：
Rampazzo, Alessandra