Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs.
Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs.
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DOI：
10.1002/ana.21627
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发表时间：
2009-06
影响因子：
11.2
通讯作者：
Hoffman E
中科院分区：
文献类型：
--
作者：
Yokota T;Lu QL;Partridge T;Kobayashi M;Nakamura A;Takeda S;Hoffman E
Duchenne肌营养不良症(DMD)是由于肌纤维膜不能产生肌营养不良蛋白引起的。一种被称为‘外显子跳跃’的反义寡核苷酸挽救Dstrophin产生的方法已被报道用于MDX小鼠和4名DMD患者的局部肌肉注射。我们试图在DMD犬模型上测试静脉注射寡核苷酸(吗啡)诱导的外显子跳跃的有效性和毒性。我们在原代细胞培养和两种体内给药方法(肌肉注射和全身静脉注射)中分别测试了一系列反义药物和鸡尾酒。多外显子跳跃(外显子6-9)的效率和有效性从mRNA、蛋白质、组织学和临床水平进行检测。在5-22周的过程中，每周或每两周一次的全身静脉注射三种吗啡诺鸡尾酒可以诱导整个身体的抗肌营养不良蛋白表达达到治疗性水平，平均约为正常水平的26%。伴随而来的是MRI和组织学检查的炎症信号减少，改善或稳定了定时跑步试验和临床症状。血液测试表明没有毒性的证据。这是首次报道在DMD的犬模型中广泛地将dystrophin的表达拯救到治疗水平。本研究也为系统性多外显子跳跃治疗提供了概念证明。使用鸡尾酒的吗啡，如图所示，允许这种方法在更大比例的DMD患者(90%)中得到更广泛的应用，并提供了选择缺失以优化dystrophin蛋白功能的前景。
Duchenne muscular dystrophy (DMD) is caused by the inability to produce dystrophin protein at the myofiber membrane. A method to rescue dystrophin production by antisense oligonucleotides, termed `exon-skipping', has been reported for the mdx mouse and in four DMD patients by local intramuscular injection. We sought to test efficacy and toxicity of intravenous oligonucleotide (morpholino) induced exon skipping in the DMD dog model. We tested a series of antisense drugs singly and as cocktails, both in primary cell culture, and two in vivo delivery methods (intramuscular injection, and systemic intravenous injection). The efficiency and efficacy of multi-exon skipping (exons 6-9) was tested at the mRNA, protein, histological, and clinical levels. Weekly or biweekly systemic intravenous injections with a three morpholino cocktail over the course of 5–22 weeks induced therapeutic levels of dystrophin expression throughout the body, with an average of about 26% normal levels. This was accompanied by reduced inflammatory signals examined by MRI and histology, improved or stabilized timed running tests and clinical symptoms. Blood tests indicated no evidence of toxicity. This is the first report of widespread rescue of dystrophin expression to therapeutic levels in the dog model of DMD. This study also provides a proof of concept for systemic multi-exon skipping therapy. Use of cocktails of morpholinos, as shown here, allows broader application of this approach to a greater proportion of DMD patients (90%), and also offers the prospect of selecting deletions that optimize the functionality of the dystrophin protein.
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影响因子：
5.1
作者：
McClorey, G.;Moulton, H. M.;Wilton, S. D.
通讯作者：
Wilton, S. D.
影响因子：
3.5
作者：
Dunckley, MG;Manoharan, M;Dickson, G
通讯作者：
Dickson, G
影响因子：
82.9
作者：
Alter, J;Lou, F;Lu, QL
通讯作者：
Lu, QL
影响因子：
2
作者：
Nakamura, Akinorl;Yoshida, Kunihiro;Ikeda, Shu-ichi
通讯作者：
Ikeda, Shu-ichi
DOI：
10.1186/1479-0556-4-3
发表时间：
2006-05-24
期刊：
Genetic vaccines and therapy
影响因子：
--
作者：
Fall, Abbie M;Johnsen, Russell;Wilton, Stephen D
通讯作者：
Wilton, Stephen D