Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells.
Early pathogenesis of Duchenne muscular dystrophy modelled in patient-derived human induced pluripotent stem cells.
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DOI：
10.1038/srep12831
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发表时间：
2015-08-20
影响因子：
4.6
通讯作者：
Sehara-Fujisawa A
中科院分区：
文献类型：
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作者：
Shoji E;Sakurai H;Nishino T;Nakahata T;Heike T;Awaya T;Fujii N;Manabe Y;Matsuo M;Sehara-Fujisawa A
Duchenne肌营养不良症(DMD)是一种由肌营养不良蛋白缺乏引起的进行性和致命性肌肉退行性疾病。有效抑制DMD的原发病理是治疗的关键。患者来源的人诱导多能干细胞(HiPSCs)是一种很有前途的药物发现工具。在这里，我们报告了一种使用HiPSCs的DMD治疗的体外评估系统，该系统概括了DMD的主要病理，并可用于DMD药物的筛选。由HiPSCs产生的骨骼肌管是完整的，这使得它们可以用于在体外模拟DMD的初始病理。诱导对照和DMD肌管在形态和生理上具有可比性。然而，电刺激这些肌管进行体外收缩，只在DMD肌细胞中引起明显的钙离子(Ca~(2+))内流。通过外显子跳跃技术恢复dystrophin可抑制DMD肌管的这种钙溢出，并减少肌酸激酶(CK)的分泌。这些结果表明，DMD的早期发病机制可以在患者来源的IPSCs诱导的骨骼肌管中有效地模拟，从而使新药的开发和评估成为可能。
Duchenne muscular dystrophy (DMD) is a progressive and fatal muscle degenerating disease caused by a dystrophin deficiency. Effective suppression of the primary pathology observed in DMD is critical for treatment. Patient-derived human induced pluripotent stem cells (hiPSCs) are a promising tool for drug discovery. Here, we report an in vitro evaluation system for a DMD therapy using hiPSCs that recapitulate the primary pathology and can be used for DMD drug screening. Skeletal myotubes generated from hiPSCs are intact, which allows them to be used to model the initial pathology of DMD in vitro. Induced control and DMD myotubes were morphologically and physiologically comparable. However, electric stimulation of these myotubes for in vitro contraction caused pronounced calcium ion (Ca2+) influx only in DMD myocytes. Restoration of dystrophin by the exon-skipping technique suppressed this Ca2+ overflow and reduced the secretion of creatine kinase (CK) in DMD myotubes. These results suggest that the early pathogenesis of DMD can be effectively modelled in skeletal myotubes induced from patient-derived iPSCs, thereby enabling the development and evaluation of novel drugs.