Intravenous infusion of an antisense oligonucleotide results in exon skipping in muscle dystrophin mRNA of Duchenne muscular dystrophy
Intravenous infusion of an antisense oligonucleotide results in exon skipping in muscle dystrophin mRNA of Duchenne muscular dystrophy
复制标题
DOI：
10.1203/01.pdr.0000215047.51278.7c
复制
发表时间：
2006-05-01
影响因子：
3.6
通讯作者：
Matsuo, M
中科院分区：
文献类型：
--
作者：
Takeshima, Y;Yagi, M;Matsuo, M
杜氏肌营养不良症（DMD）是一种以肌营养不良蛋白缺乏为特征的致命性肌肉萎缩性疾病。我们报告，静脉（IV）输注的反义寡核苷酸创建一个框内肌营养不良蛋白mRNA从框外DMD突变（通过外显子跳跃），导致肌肉肌营养不良蛋白的表达。一名10岁的DMD患者，其具有肌营养不良蛋白基因的框外外显子20缺失，接受0.5 mg/kg IV输注针对外显子19的剪接增强子序列的反义31-mer硫代磷酸酯寡核苷酸。该反义构建体以一周的间隔施用4周。未观察到归因于输注的副作用。输注后，外周淋巴细胞中抗肌萎缩蛋白mRNA的一部分出现外显子19跳跃。在最后一次输注后一周的肌肉活组织检查中，鉴定了缺乏外显子19和20的新型框内mRNA，发现其约占总逆转录PCR产物的6%。肌营养不良蛋白的寡核苷酸处理后的肌细胞的肌膜中的组织化学鉴定。这些发现表明，硫代磷酸寡核苷酸可以安全地给予DMD儿童，并且简单的IV输注是导致DMD骨骼肌中外显子跳跃的寡核苷酸的有效递送机制。
Duchenne Muscular dystrophy (DMD) is a fatal muscle wasting disease that is characterized by muscle dystrophin deficiency. We report that intravenous (IV) infusion of an antisense oligonucleotide created an in-frame dystrophin mRNA from an out-of-frame DMD mutation (via exon skipping) which led to muscle dystrophin expression. A 10-year-old DMD patient possessing an out-of-frame, exon 20 deletion of the dystrophin gene received a 0.5 mg/kg IV infusion of an antisense 31-mer phosphorothioate oligonucleotide against the splicing enhancer sequence of exon 19. This antisense construct was administered at one-week intervals for 4 wk. No side effects attributable to infusion were observed. Exon 19 skipping appeared in a portion of the dystrophin mRNA in peripheral lymphocytes after the infusion. In a muscle biopsy one week after the final infusion, the novel in-frame mRNA lacking both exons 19 and 20 was identified and found to represent approximately 6% of the total reverse transcription PCR product. Dystrophin was identified histochemically in the sarcolemma of muscle cells after oligonucleotide treatment. These findings demonstrate that phosphorothioate oligonucleotides may be administered safely to children with DMD, and that a simple IV infusion is an effective delivery mechanism for oligonucleotides that lead to exon skipping in DMD skeletal muscles.