Dose-dependent restoration of dystrophin expression in cardiac muscle of dystrophic mice by systemically delivered morpholino
Dose-dependent restoration of dystrophin expression in cardiac muscle of dystrophic mice by systemically delivered morpholino
复制标题
DOI：
10.1038/gt.2009.120
复制
发表时间：
2010-01-01
期刊：
影响因子：
5.1
通讯作者：
Lu, Q. L.
中科院分区：
文献类型：
--
作者：
Wu, B.;Lu, P.;Lu, Q. L.
我们之前已经证明反义morpholino寡聚物能够通过全身递送在营养不良的mdx小鼠的全身骨骼肌中恢复肌营养不良蛋白的表达。然而，肌营养不良蛋白的表达水平差异很大，更重要的是，心肌中没有肌营养不良蛋白的表达。在这项研究中，我们通过剂量递增的方法研究了morpholino诱导的外显子跳跃在体外心肌细胞和心肌细胞中的效率。我们发现，morpholino在骨骼肌成肌细胞和心肌细胞中同样有效地诱导靶向外显子跳跃。在培养的心肌细胞中实现了有效的外显子跳跃。在mdx小鼠中，morpholino对骨骼肌和心肌中肌营养不良蛋白的表达均有剂量依赖性。心肌中的肌营养不良蛋白达到了治疗水平，尽管其剂量高于骨骼肌。单次全身递送3 g kg(-1) morpholino在骨骼肌和心肌中分别诱导高达50%和30%正常水平的肌营养不良蛋白。高剂量的morpholino治疗降低了血清肌酸激酶的水平，但没有明显的毒性。这些结果表明，在杜氏肌营养不良的治疗中，morpholino可以有效地挽救心肌中的肌营养不良蛋白。基因治疗（2010）17,132-140；doi: 10.1038 / gt.2009.120;2009年9月17日在线发布
We have earlier shown that antisense morpholino oligomers are able to restore dystrophin expression by systemic delivery in body-wide skeletal muscles of dystrophic mdx mice. However, the levels of dystrophin expression vary considerably and, more importantly, no dystrophin expression has been achieved in cardiac muscle. In this study, we investigate the efficiency of morpholino-induced exon skipping in cardiomyoblasts and myocytes in vitro, and in cardiac muscle in vivo by dose escalation. We showed that morpholino induces targeted exon skipping equally effectively in both skeletal muscle myoblasts and cardiomyoblasts. Effective exon skipping was achieved in cardiomyocytes in culture. In the mdx mice, morpholino rescues dystrophin expression dose dependently in both skeletal and cardiac muscles. Therapeutic levels of dystrophin were achieved in cardiac muscle albeit at higher doses than in skeletal muscles. Up to 50 and 30% normal levels of dystrophin were induced by single systemic delivery of 3 g kg(-1) of morpholino in skeletal and cardiac muscles, respectively. High doses of morpholino treatment reduced the serum levels of creatine kinase without clear toxicity. These findings suggest that effective rescue of dystrophin in cardiac muscles can be achieved by morpholino for the treatment of Duchenne muscular dystrophy. Gene Therapy (2010) 17, 132-140; doi: 10.1038/gt.2009.120; published online 17 September 2009