Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN 55455, USA.
Stem Cell Institute, University of Minnesota, Minneapolis, MN 55455, USA.
Cells. 2024 Jun 4;13(11):972. doi: 10.3390/cells13110972.
Mutations in the gene cause fatal Duchenne Muscular Dystrophy (DMD). An attractive therapeutic approach is autologous cell transplantation utilizing myogenic progenitors derived from induced pluripotent stem cells (iPSCs). Given that a significant number of DMD mutations occur between exons 45 and 55, we developed a gene knock-in approach to correct any mutations downstream of exon 44. We applied this approach to two DMD patient-specific iPSC lines carrying mutations in exons 45 and 51 and confirmed mini-DYSTROPHIN (mini-DYS) protein expression in corrected myotubes by western blot and immunofluorescence staining. Transplantation of gene-edited DMD iPSC-derived myogenic progenitors into NSG/mdx mice produced donor-derived myofibers, as shown by the dual expression of human DYSTROPHIN and LAMIN A/C. These findings further provide proof-of-concept for the use of programmable nucleases for the development of autologous iPSC-based therapy for muscular dystrophies.
基因中的突变会导致致命的杜氏肌营养不良症（DMD）。一种有吸引力的治疗方法是利用诱导多能干细胞（iPSC）衍生的成肌祖细胞进行自体细胞移植。鉴于大量的 DMD 突变发生在 45 号和 55 号外显子之间，我们开发了一种基因敲入方法来纠正 44 号外显子下游的任何突变。我们将这种方法应用于携带 45 号和 51 号外显子突变的两个 DMD 患者特异性 iPSC 系，并通过 Western blot 和免疫荧光染色证实校正后的肌管中存在 mini-DYSTROPHIN（mini-DYS）蛋白表达。将基因编辑的 DMD iPSC 衍生的成肌祖细胞移植到 NSG/mdx 小鼠中，产生了供体来源的肌纤维，如人 DYSTROPHIN 和 LAMIN A/C 的双重表达所示。这些发现进一步为使用可编程核酸酶开发用于肌肉营养不良的自体 iPSC 为基础的治疗提供了概念验证。
