2'-O-Methyl RNA/Ethylene-Bridged Nucleic Acid Chimera Antisense Oligonucleotides to Induce Dystrophin Exon 45 Skipping.
2'-O-Methyl RNA/Ethylene-Bridged Nucleic Acid Chimera Antisense Oligonucleotides to Induce Dystrophin Exon 45 Skipping.
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DOI：
10.3390/genes8020067
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发表时间：
2017-02-10
期刊：
影响因子：
3.5
通讯作者：
Matsuo M
中科院分区：
文献类型：
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作者：
Lee T;Awano H;Yagi M;Matsumoto M;Watanabe N;Goda R;Koizumi M;Takeshima Y;Matsuo M
杜氏肌营养不良症（DMD）是一种致命的肌肉萎缩疾病，其特征是由肌营养不良蛋白基因突变引起的肌营养不良蛋白缺乏。反义寡核苷酸（AO）介导的外显子跳变旨在恢复肌营养不良蛋白阅读框，从而允许产生内部缺失的肌营养不良蛋白，对框架外缺失的DMD患者具有功能益处。在美国加速批准eteplirsen （Exondys 51）后，现在的工作重点是靶向其他外显子。eteplirsen靶向肌营养不良蛋白51外显子。为了提高临床疗效，这一策略需要对核酸的递送方法和修饰进行更多的研究。我们研究了一种具有2 ' -O,4 ' - c -乙烯桥接核酸（ENA）的核苷酸，该核苷酸具有高核酸酶抗性和对互补RNA链的高亲和力。在这里，我们描述了开发2 ' - o -甲基RNA(2 ' -OMeRNA)/ENA嵌合体AO诱导肌营养不良蛋白外显子45跳变的过程。一个18-mer 2 ' -OMeRNA/ENA嵌合体（AO85）在培养的肌管中诱导45外显子跳跃的活性最强。AO85用于mdx小鼠，无明显副作用。将AO85转染到13例DMD患者的培养肌管中，所有样本的45外显子均有不同程度的跳跃，11例患者的肌营养不良蛋白表达。这些结果表明，AO介导的外显子跳变在个体患者中可能有效，并突出了2 ' -OMeRNA/ENA嵌合AO作为DMD潜在的基础治疗方法。
Duchenne muscular dystrophy (DMD) is a fatal muscle-wasting disease characterized by dystrophin deficiency from mutations in the dystrophin gene. Antisense oligonucleotide (AO)-mediated exon skipping targets restoration of the dystrophin reading frame to allow production of an internally deleted dystrophin protein with functional benefit for DMD patients who have out-of-frame deletions. After accelerated US approval of eteplirsen (Exondys 51), which targets dystrophin exon 51 for skipping, efforts are now focused on targeting other exons. For improved clinical benefits, this strategy requires more studies of the delivery method and modification of nucleic acids. We studied a nucleotide with a 2′-O,4′-C-ethylene-bridged nucleic acid (ENA), which shows high nuclease resistance and high affinity for complementary RNA strands. Here, we describe the process of developing a 2′-O-methyl RNA(2′-OMeRNA)/ENA chimera AO to induce dystrophin exon 45 skipping. One 18-mer 2′-OMeRNA/ENA chimera (AO85) had the most potent activity for inducing exon 45 skipping in cultured myotubes. AO85 was administered to mdx mice without significant side effects. AO85 transfection into cultured myotubes from 13 DMD patients induced exon 45 skipping in all samples at different levels and dystrophin expression in 11 patients. These results suggest the possible efficacy of AO-mediated exon skipping changes in individual patients and highlight the 2′-OMeRNA/ENA chimera AO as a potential fundamental treatment for DMD.