Autor: |
Hiroyuki Inoue, Satoki Nakamura, Shuichiro Higo, Mikio Shiba, Yasuaki Kohama, Takumi Kondo, Satoshi Kameda, Tomoka Tabata, Shota Okuno, Yoshihiko Ikeda, Junjun Li, Li Liu, Satoru Yamazaki, Maki Takeda, Emiko Ito, Seiji Takashima, Shigeru Miyagawa, Yoshiki Sawa, Shungo Hikoso, Yasushi Sakata |
Rok vydání: |
2020 |
Předmět: |
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Zdroj: |
Stem cell reports. 17(2) |
ISSN: |
2213-6711 |
Popis: |
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. |
Databáze: |
OpenAIRE |
Externí odkaz: |
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