Development of Human Disease Model of Arrhythmogenic Right Ventricular Cardiomyopathy for Personalized Medicine

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21602
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 53:Organ-based internal medicine and related fields
• Research Institution: Osaka University
• Principal Investigator: Sakata Yasushi 大阪大学, 医学系研究科, 教授 (00397671)
• Co-Investigator(Kenkyū-buntansha): 肥後 修一朗 大阪大学, 医学系研究科, 特任准教授(常勤) (00604034) ; 彦惣 俊吾 大阪大学, 医学系研究科, 准教授 (30423164)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: 不整脈原性心筋症 / 疾患iPS細胞 / ゲノム編集

Outline of Final Research Achievements

Loss-of-function mutations in PKP2, encoding plakophilin-2, cause arrhythmogenic cardiomyopathy (AC). 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 in the generated isogenic lines enabled real-time desmosome-imaging under an adjusted dose of plakophilin-2. AAV-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.

Free Research Field

循環器内科学

Academic Significance and Societal Importance of the Research Achievements

不整脈原性心筋症には既存の心不全治療薬が奏功せず、進行した場合、心臓移植しか治療方法が残されていません。本研究ではプラコフィリン2タンパク質が欠損したiPS細胞由来分化心筋細胞を用いて、その病態解明を行い、遺伝子補充の治療概念を実証しました。精密に遺伝子を改変した不整脈原性心筋症モデル細胞、及びデスモゾームを可視化するイメージング細胞は、難治性心筋症に対する今後の治療法開発において、有用なプラットフォームを提供すると考えられます。