Modeling Duchenne muscular dystrophy using patient-derived iPS cells

使用患者来源的 iPS 细胞模拟杜氏肌营养不良症

• 批准号: 24790383
• 负责人: SAKURAI Hidetoshi
• 金额: $ 2.91万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2012
• 资助国家: 日本
• 起止时间: 2012-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-24790383/
• 关键词: iPS細胞; 疾患再現; 筋ジストロフィー; デュシェンヌ型筋ジストロフィー; 細胞内カルシウムイメージング; 疾患特異的iPS細胞; 病態再現; 骨格筋細胞; 創薬スクリーニング

In this study, we demonstrated the early pathogenesis of Duchenne muscular dystrophy (DMD) using patient-derived iPS cells. DMD-muscles differentiated from DMD-iPSCs exhibited similar gene expression profile and similar maturity as control iPSC-derived muscles, and no differentiation delay was observed in DMD-muscles. When DMD-muscles were exposed to electric stimulation, higher intracellular Ca2+ influx was observed in DMD-muscles than control-muscles. Larger cell damage was observed in DMD-muscles than control-muscles through overloading Ca2+ by ionomycin. Since these phenotype could be ameliorated by restored dystrophin expression by exon skip technics, we conclude that excessive Ca2+ influx is the trigger for the early pathogenesis of DMD caused by absence of dystrophin. According to our findings, we are going to develop new drugs targeting excessive Ca2+ influx by analyzing the mechanism which is caused by absence of dystrophin.

在这项研究中，我们证明了杜氏肌营养不良症（DMD）的早期发病机制，使用患者来源的iPS细胞。从DMD-iPSC分化的DMD肌肉表现出与对照iPSC衍生的肌肉相似的基因表达谱和相似的成熟度，并且在DMD肌肉中没有观察到分化延迟。当DMD肌肉暴露于电刺激时，DMD肌肉中观察到比对照肌肉更高的细胞内Ca 2+内流。离子霉素使DMD肌细胞钙超载，细胞损伤程度明显大于对照肌。由于这些表型可以通过外显子跳跃技术恢复肌营养不良蛋白的表达来改善，我们得出结论，过量的Ca 2+内流是肌营养不良蛋白缺失引起的DMD早期发病的触发因素。根据我们的研究结果，我们将通过分析肌营养不良蛋白缺乏引起的机制来开发针对过度Ca 2+内流的新药。

项目成果

デュシェンヌ型筋ジストロフィー患者由来iPS細胞を用いての病態再現

使用源自杜氏肌营养不良症患者的 iPS 细胞再现病理状况

• 作者: 庄子栄美;櫻井英俊;中畑龍俊;田中章仁;ウォルツェン クヌート;藤井宣晴;平家敏男;真鍋康子;瀬原-藤沢淳子
• 通讯作者: 瀬原-藤沢淳子