Modeling reduced contractility and impaired desmosome assembly due to plakophilin-2 deficiency using isogenic iPS cell-derived cardiomyocytes.
Modeling reduced contractility and impaired desmosome assembly due to plakophilin-2 deficiency using isogenic iPS cell-derived cardiomyocytes.
复制标题
DOI：
10.1016/j.stemcr.2021.12.016
复制
发表时间：
2022-02-08
影响因子：
5.9
通讯作者：
Sakata Y
中科院分区：
文献类型：
--
作者：
Inoue H;Nakamura S;Higo S;Shiba M;Kohama Y;Kondo T;Kameda S;Tabata T;Okuno S;Ikeda Y;Li J;Liu L;Yamazaki S;Takeda M;Ito E;Takashima S;Miyagawa S;Sawa Y;Hikoso S;Sakata Y
PKP 2（编码斑嗜蛋白-2）的功能缺失突变可导致致突变性心肌病（AC）。缺陷分子的恢复可以用作上游疗法，从而需要一种人类模型，其重现疾病病理学并在表型分析中提供不同的读数，用于基因替代疗法的概念证明。在这里，我们从携带杂合移码PKP 2突变的AC患者中产生了具有精确调节的plakophilin-2表达的同基因诱导多能干细胞衍生的心肌细胞（iPSC-CM）。在单层分化后，plakophilin-2缺乏导致iPSC-CM中收缩性降低、闰盘结构破坏和桥粒组装受损。等位基因特异性荧光标记的内源性DSG 2编码桥粒芯糖蛋白-2在所产生的等基因系，使实时桥粒成像下调整剂量的斑嗜蛋白-2。腺相关病毒介导的PKP 2基因置换恢复了收缩性并恢复了桥粒组装，这在plakophilin-2缺陷的iPSC-CM中通过桥粒成像顺序捕获。我们的iPSC-CM的同基因组概括了AC病理学，并为治疗开发提供了快速方便的细胞平台。使用精确剂量的plakophilin-2生成等基因iPSC-CM使用iPSC-CM建模收缩性降低和桥粒组装受损等基因iPSC-CM用于桥粒成像的生成等基因iPSC-CM使用等基因plakophilin-2缺陷iPSC-CM替代PKP 2的概念验证在这篇文章中，Higo及其同事生成了等基因iPS细胞衍生的心肌细胞在具有精确调节的PKP 2表达的iPSC-CM中，建模由于斑嗜蛋白-2缺乏而降低的收缩性和受损的桥粒组装。PKP 2基因置换恢复了收缩性并恢复了桥粒组装，这被桥粒成像顺序地捕获。同基因iPSC-CM概括了致瘤性心肌病的病理学，并为治疗开发提供了快速方便的细胞平台。
Loss-of-function mutations in PKP2, which encodes plakophilin-2, cause arrhythmogenic cardiomyopathy (AC). Restoration of deficient molecules can serve as upstream therapy, thereby requiring a human model that recapitulates disease pathology and provides distinct readouts in phenotypic analysis for proof of concept for gene replacement therapy. Here, we generated isogenic induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) with precisely adjusted expression of plakophilin-2 from a patient with AC carrying a heterozygous frameshift PKP2 mutation. After monolayer differentiation, plakophilin-2 deficiency led to reduced contractility, disrupted intercalated disc structures, and impaired desmosome assembly in iPSC-CMs. Allele-specific fluorescent labeling of endogenous DSG2 encoding desmoglein-2 in the generated isogenic lines enabled real-time desmosome-imaging under an adjusted dose of plakophilin-2. Adeno-associated virus-mediated gene replacement of PKP2 recovered contractility and restored desmosome assembly, which was sequentially captured by desmosome-imaging in plakophilin-2-deficient iPSC-CMs. Our isogenic set of iPSC-CMs recapitulates AC pathology and provides a rapid and convenient cellular platform for therapeutic development. Generation of isogenic iPSC-CMs with a precise dose of plakophilin-2 Modeling reduced contractility and impaired desmosome assembly using iPSC-CMs Generation of isogenic iPSC-CMs for desmosome-imaging Proof of concept of PKP2 replacement using isogenic plakophilin-2-deficient iPSC-CMs In this article, Higo and colleagues generate isogenic iPS cell-derived cardiomyocytes (iPSC-CMs) with precisely adjusted expression of PKP2, modeling reduced contractility and impaired desmosome assembly due to plakophilin-2 deficiency. PKP2 gene replacement recovered contractility and restored desmosome assembly, which was sequentially captured by desmosome imaging. The isogenic iPSC-CMs recapitulate the pathology of arrhythmogenic cardiomyopathy and provide a rapid and convenient cellular platform for therapeutic development.
登录
查看更多内容
通过非分裂心肌细胞中的同源指导修复靶向基因组替代。
影响因子：
4.6
作者：
Ishizu T;Higo S;Masumura Y;Kohama Y;Shiba M;Higo T;Shibamoto M;Nakagawa A;Morimoto S;Takashima S;Hikoso S;Sakata Y
通讯作者：
Sakata Y
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：
64.8
作者：
通讯作者：
--
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：
3.5
作者：
Ramond, F.;Janin, A.;Millat, G.
通讯作者：
Millat, G.