Engraftment of human induced pluripotent stem cell-derived myogenic progenitors restores dystrophin in mice with duchenne muscular dystrophy
Engraftment of human induced pluripotent stem cell-derived myogenic progenitors restores dystrophin in mice with duchenne muscular dystrophy
复制标题
人类诱导多能干细胞来源的肌源性祖细胞的植入可恢复杜氏肌营养不良症小鼠的肌营养不良蛋白
DOI：
10.1186/s40659-020-00288-1
复制
发表时间：
2020-01-01
影响因子：
6.7
通讯作者：
Zhang, Cheng
中科院分区：
文献类型：
--
作者：
He, Ruojie;Li, Huan;Zhang, Cheng
研究背景杜氏肌营养不良症（DMD）是一种遗传性肌肉疾病，由于dystrophin基因的缺失，目前尚无有效的治疗方法。人类诱导多能干细胞（hiPSC）为肌营养不良症的细胞疗法提供了有前途的无限资源。然而，它们的临床应用受到低效的肌源性分化的阻碍，此外，尚未在DMD的mdx小鼠模型中检查非转基因hiPSC衍生的肌源性祖细胞的植入。方法研究mdx小鼠hiPSCs来源的肌源性祖细胞的肌肉再生能力。用增强型绿色荧光蛋白（EGFP）载体转染hiPSCs，并将其定义为EGFP hiPSCs。在碱性成纤维细胞生长因子、毛喉素、6-溴靛玉红-3 '-肟以及马血清的补充下，在EGFP hiPSCs上进行成肌分化。通过肌内和静脉内注射将EGFP hiPSC衍生的肌源性祖细胞移植到mdx小鼠中。观察肌内和全身移植后肌营养不良蛋白表达的恢复情况、中央核肌纤维的比例以及移植细胞来源的卫星细胞。结果我们报道了用这三种小分子处理后，hiPSCs可以产生大量的肌源性祖细胞，随后在体外终末分化产生成熟的肌管。肌内或全身移植到mdx小鼠后，这些肌源性祖细胞移植并促进宿主肌肉中的人源性肌纤维再生，恢复肌营养不良蛋白表达，改善病理损伤，并在营养不良的肌肉中接种卫星细胞区室。结论非转基因诱导hiPSCs来源的肌源性祖细胞具有肌肉再生潜能。hiPSC衍生的肌源性祖细胞的移植可能是在临床环境中治疗DMD的潜在未来治疗策略。
Background Duchenne muscular dystrophy (DMD) is a devastating genetic muscular disorder with no effective treatment that is caused by the loss of dystrophin. Human induced pluripotent stem cells (hiPSCs) offer a promising unlimited resource for cell-based therapies of muscular dystrophy. However, their clinical applications are hindered by inefficient myogenic differentiation, and moreover, the engraftment of non-transgene hiPSC-derived myogenic progenitors has not been examined in the mdx mouse model of DMD. Methods We investigated the muscle regenerative potential of myogenic progenitors derived from hiPSCs in mdx mice. The hiPSCs were transfected with enhanced green fluorescent protein (EGFP) vector and defined as EGFP hiPSCs. Myogenic differentiation was performed on EGFP hiPSCs with supplementary of basic fibroblast growth factor, forskolin, 6-bromoindirubin-3 '-oxime as well as horse serum. EGFP hiPSCs-derived myogenic progenitors were engrafted into mdx mice via both intramuscular and intravenous injection. The restoration of dystrophin expression, the ratio of central nuclear myofibers, and the transplanted cells-derived satellite cells were accessed after intramuscular and systemic transplantation. Results We report that abundant myogenic progenitors can be generated from hiPSCs after treatment with these three small molecules, with consequent terminal differentiation giving rise to mature myotubes in vitro. Upon intramuscular or systemic transplantation into mdx mice, these myogenic progenitors engrafted and contributed to human-derived myofiber regeneration in host muscles, restored dystrophin expression, ameliorated pathological lesions, and seeded the satellite cell compartment in dystrophic muscles. Conclusions This study demonstrates the muscle regeneration potential of myogenic progenitors derived from hiPSCs using non-transgenic induction methods. Engraftment of hiPSC-derived myogenic progenitors could be a potential future therapeutic strategy to treat DMD in a clinical setting.