Elucidation of molecular mechanism of utrophin expression in dystrophic skeletal muscle and its application to molecular therapy

营养不良性骨骼肌中utropin表达的分子机制的阐明及其在分子治疗中的应用

基本信息

批准号：
11470153
负责人：
TAKEDA Shin'ichi
金额：
$ 9.54万
依托单位：
National Institute of Neuroscience, NCNP
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470153/
关键词：
utrophin dystrophin mdx mice DMD adenovirus vector stabilization of mRNA cytokines IL-6 免疫応答炎症反応免疫反応

项目摘要

Duchenne muscular dystrophy (DMD) is an X-linked lethal disorder caused by a defect in the DMD gene, which encodes the cytoskeletal protein dystrophin. Utrophin is an autosomal homologue of the DMD gene product dystrophin, and augmented expression of endogenous utrophin is expected to provide an alternative therapeutic approach for DMD. We previously reported that immune response against a (β-galactosidase-expressing adenovirus vector, AxCALacZ, resulted in accumulation of endogenous utrophin expression on the extrasynaptic sarcolemma in dystrophin-deficient mdx mice (Hum Gene Ther 11 : 669-680, 2000). In order to determine which cytokine is involved in the regulation of utrophin expression, we directly injected several cytokines separately into neonatal mdx muscles and tested whether the expression of utrophin is increased on the sarcolemma or not. Importantly, among the cytokines tested, solely IL-6 successfully increased expression of utrophin. Moreover, increase of utrophin mRNA was detected from rIL-6-injected mdx muscles by the quantitative real-time RT-PCR study. Further, IL-6 expression was elevated in AxCALacZ-infected mdx muscle in the early stage, and anti-IL-6R antibody treatment blocked enhanced utrophin expression in AxCALacZinfected mdx muscle. We should point out, however, that over-expression of utrophin due to recombinant IL-6 treatment lasted only a week. In addition, expression of utrophin was not evident in normal C57BL/10 neonatal muscles injected with IL-6 (Hum Gene Ther 13 : 509-518, 2002). Taken together, these results suggest that IL6 can induce over-expression of utrophin on the extrasynaptic sarcolemma but requires pre-existing factors in neonatal mdx muscle to fully regulate utrophin expression.

Duchenne肌肉营养不良症（DMD）是由DMD基因缺陷引起的X连锁致命疾病，该缺陷编码了细胞骨架蛋白肌营养不良蛋白。 utrophin是DMD基因产物肌营养不良蛋白的常染色体同源物，预计内源性肌营养不良蛋白的表达增强将为DMD提供另一种治疗方法。 We previously reported that immune response against a (β-galactosidase-expressing adenovirus vector, AxCALacZ, resulted in accumulation of endogenous sarcolemma in dystrophin-deficient mdx mice (Hum Gene Ther 11: 669-680, 2000). In order to determine which cytokine is involved in the regulation of utrophin expression, we directly injected several cytokines separately新生儿MDX的肌肉和肌膜上的表达是否在肌膜上增加，在测试的细胞因子中，IL-6成功地增加了utrophin的表达。在早期的Axcalacz感染的MDX肌肉，抗IL-6R抗体处理在轴心唱的MDX肌肉中阻止了乌托蛋白的表达，我们应该指出，由于IL-6治疗持续了10次，我们的表达效果不佳。注射IL-6（Hum Gene ther 13：509-518，2002），这些结果表明，IL6可以诱导乌托蛋白过度表达在外鼻刺激性的肌膜上，但需要在新生儿MDX肌肉中固定的因素才能完全调节utrophin的表达。