Single Intramuscular Injection of AAV-shRNA Reduces DNM2 and Prevents Myotubular Myopathy in Mice
Single Intramuscular Injection of AAV-shRNA Reduces DNM2 and Prevents Myotubular Myopathy in Mice
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DOI：
10.1016/j.ymthe.2018.02.008
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发表时间：
2018-04-04
影响因子：
12.4
通讯作者：
Cowling, Belinda S.
中科院分区：
文献类型：
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作者：
Tasfaout, Hichem;Lionello, Valentina M.;Cowling, Belinda S.
肌小管肌病，或x连锁核中心肌病，是一种严重的肌肉疾病，对患者及其家属来说是一个重大的负担。它的临床特征是新生儿和严重的肌肉无力和萎缩。肌小管蛋白（MTM1）基因突变可引起肌小管肌病，目前尚无特异性治疗方法。我们之前发现，在Mtm1敲除和患者肌肉样本中，动力蛋白2 （DNM2）都上调，而通过反义寡核苷酸减少动力蛋白2 (DNM2)，可以挽救小鼠这种肌病的临床和组织病理学特征。在这里，我们提出了一种针对Dnm2 mRNA的新方法。我们在体外和体内筛选并验证了几种有效靶向Dnm2 mRNA的短发夹RNA （shRNA）序列。单次肌内注射AAV-shDnm2可长期降低DNM2蛋白水平，恢复肌肉力量、质量、组织学和肌纤维超微结构，并预防与疾病相关的分子缺陷。我们的研究结果证明了DNM2的强大敲除，并提供了一种基于DNM2减少治疗肌小管肌病的替代策略。
Myotubular myopathy, or X-linked centronuclear myopathy, is a severe muscle disorder representing a significant burden for patients and their families. It is clinically characterized by neonatal and severe muscle weakness and atrophy. Mutations in the myotubularin (MTM1) gene cause myotubular myopathy, and no specific curative treatment is available. We previously found that dynamin 2 (DNM2) is upregulated in both Mtm1 knockout and patient muscle samples, whereas its reduction through antisense oligonucleotides rescues the clinical and histopathological features of this myopathy in mice. Here, we propose a novel approach targeting Dnm2 mRNA. We screened and validated in vitro and in vivo several short hairpin RNA (shRNA) sequences that efficiently target Dnm2 mRNA. A single intramuscular injection of AAV-shDnm2 resulted in long-term reduction of DNM2 protein level and restored muscle force, mass, histology, and myofiber ultrastructure and prevented molecular defects linked to the disease. Our results demonstrate a robust DNM2 knockdown and provide an alternative strategy based on reduction of DNM2 to treat myotubular myopathy.