Autologous transplantation of SM/C-2.6+ satellite cells transduced with micro-dystrophin CS1 cDNA by lentiviral vector into mdx mice
Autologous transplantation of SM/C-2.6+ satellite cells transduced with micro-dystrophin CS1 cDNA by lentiviral vector into mdx mice
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DOI：
10.1038/sj.mt.6300295
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发表时间：
2007-12-01
影响因子：
12.4
通讯作者：
Takeda, Shin'ichi
中科院分区：
文献类型：
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作者：
Ikemoto, Madoka;Fukada, So-ichiro;Takeda, Shin'ichi
杜氏肌营养不良症（DMD）是由肌营养不良蛋白基因突变引起的致死性肌肉疾病。移植自体肌源性细胞遗传校正离体是一种可能的治疗这种疾病。为了测试新鲜分离的卫星细胞的再生效率，我们使用SM/C-2.6（最近开发的单克隆抗体）和流式细胞术从8-12周龄的绿色荧光蛋白转基因（GFP-Tg）小鼠的四肢肌肉中纯化静止的卫星细胞。当移植到8-12周龄心脏毒素注射的C57 BL/6小鼠和5周龄肌营养不良蛋白缺陷型mdx小鼠的胫骨前肌（TA）肌肉中时，新鲜分离的卫星细胞显示出比卫星细胞衍生的成肌细胞更有效地参与肌肉再生，并在注射后4周进行分析。重要的是，在体外扩增新鲜分离的卫星细胞而不传代对其再生能力没有不利影响。因此，我们使用SM/C-2.6抗体直接从5周龄mdx小鼠中分离卫星细胞，并将其与表达微小肌养蛋白CS 1的慢病毒载体一起培养。将转导的细胞注射到5周龄mdx小鼠的TA肌肉中。移植后4周，移植细胞有效地促进mdx营养不良肌肉的再生，并在肌膜上表达微肌营养不良蛋白。这些结果表明，慢病毒载体介导的DMD离体基因治疗是有潜力的。
Duchenne muscular dystrophy (DMD) is a lethal muscle disorder caused by mutations in the dystrophin gene. Transplantation of autologous myogenic cells genetically corrected ex vivo is a possible treatment for this disorder. In order to test the regenerative efficiency of freshly isolated satellite cells, we purified quiescent satellite cells from limb muscles of 8-12-week-old green fluorescent protein-transgenic (GFP-Tg) mice using SM/C-2.6 (a recently developed monoclonal antibody) and flow cytometry. Freshly isolated satellite cells were shown to participate in muscle regeneration more efficiently than satellite cell-derived myoblasts passaged in vitro do, when transplanted into tibialis anterior (TA) muscles of 8-12-week-old cardiotoxin-injected C57BL/6 mice and 5-week-old dystrophin-deficient mdx mice, and analyzed at 4 weeks after injection. Importantly, expansion of freshly isolated satellite cells in vitro without passaging had no detrimental effects on their regenerative capacity. Therefore we directly isolated satellite cells from 5-week-old mdx mice using SM/C-2.6 antibody and cultured them with lentiviral vectors expressing micro-dystrophin CS1. The transduced cells were injected into TA muscles of 5-week-old mdx mice. At 4 weeks after transplantation, the grafted cells efficiently contributed to regeneration of mdx dystrophic muscles and expressed micro-dystrophin at the sarcolemma. These results suggest that there is potential for lentiviral vector-mediated ex vivo gene therapy for DMD.