Antisense correction of SMN2 splicing in the CNS rescues necrosis in a type III SMA mouse model
Antisense correction of SMN2 splicing in the CNS rescues necrosis in a type III SMA mouse model
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DOI：
10.1101/gad.1941310
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发表时间：
2010-08-01
影响因子：
10.5
通讯作者：
Krainer, Adrian R.
中科院分区：
文献类型：
--
作者：
Hua, Yimin;Sahashi, Kentaro;Krainer, Adrian R.
增加运动神经元2，着丝粒（SMN 2）外显子7包含的存活以在运动神经元中表达更多全长SMN蛋白是治疗脊髓性肌萎缩症（SMA）的一种有希望的方法。之前，我们确定了一种有效的2 '-O-（2-甲氧基乙基）（莫伊）硫代磷酸酯修饰的反义寡核苷酸（阿索），可阻断SMN 2内含子剪接沉默元件，并在全身给药后有效促进转基因小鼠外周组织中的外显子7包含。在这里，我们解决其在脊髓的疗效-疾病治疗的先决条件-和它的能力，以挽救轻度SMA小鼠模型，发展尾巴和耳朵坏死，类似于在一些SMA婴儿的远端组织坏死报告。使用微渗透泵，我们将阿索直接输注到表达人SMN 2转基因的成年小鼠的侧脑室中;在脊髓运动神经元的mRNA和蛋白质水平上测量，阿索使SMN 2外显子7包涵体稳健且持久地增加。单次胚胎或新生儿侧脑室注射阿索可显著挽救SMA小鼠的尾部和耳部坏死。我们的结论是，这种莫伊阿索是SMA治疗的一种有前途的候选药物，更一般地说，ASO可用于有效地重定向CNS中靶基因的选择性剪接。
Increasing survival of motor neuron 2, centromeric (SMN2) exon 7 inclusion to express more full-length SMN protein in motor neurons is a promising approach to treat spinal muscular atrophy (SMA), a genetic neurode-generative disease. Previously, we identified a potent 2'-O-(2-methoxyethyl) (MOE) phosphorothioate-modified antisense oligonucleotide (ASO) that blocks an SMN2 intronic splicing silencer element and efficiently promotes exon 7 inclusion in transgenic mouse peripheral tissues after systemic administration. Here we address its efficacy in the spinal cord-a prerequisite for disease treatment-and its ability to rescue a mild SMA mouse model that develops tail and ear necrosis, resembling the distal tissue necrosis reported in some SMA infants. Using a micro-osmotic pump, we directly infused the ASO into a lateral cerebral ventricle in adult mice expressing a human SMN2 transgene; the ASO gave a robust and long-lasting increase in SMN2 exon 7 inclusion measured at both the mRNA and protein levels in spinal cord motor neurons. A single embryonic or neonatal intracerebroventricular ASO injection strikingly rescued the tail and ear necrosis in SMA mice. We conclude that this MOE ASO is a promising drug candidate for SMA therapy, and, more generally, that ASOs can be used to efficiently redirect alternative splicing of target genes in the CNS.