Transduction of full-length dystrophin to multiple skeletal muscles improves motor performance and life span in utrophin/dystrophin double knockout mice
Transduction of full-length dystrophin to multiple skeletal muscles improves motor performance and life span in utrophin/dystrophin double knockout mice
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DOI：
10.1038/mt.2008.23
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发表时间：
2008-05-01
影响因子：
12.4
通讯作者：
Uchino, Makoto
中科院分区：
文献类型：
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作者：
Kawano, Ryoko;Ishizaki, Masatoshi;Uchino, Makoto
杜氏肌营养不良症（DMD）是一种致命的、进行性的、肌肉萎缩性疾病，由肌营养不良蛋白缺陷引起。除了辅助病毒依赖性腺病毒载体（HDAdv）之外，没有病毒载体可以包装14-乙酰化酶（kb）全长肌营养不良蛋白互补DNA（cDNA），并且HDAdv由于其基因组中的大尺寸缺失而比老一代腺病毒载体安全得多。我们已经产生了携带myc标记的鼠全长肌营养不良蛋白cDNA（HDAdv-myc-mFLdys）的HDAdv。我们将其注射到7天大的utrophin/dystrophin双敲除小鼠（dko小鼠）的多个近端肌肉中（这些小鼠通常表现出与人类DMD非常相似的症状），因为DMD患者的近端肌肉受到影响。注射8周后，转导的肌营养不良蛋白广泛表达，我们发现中央有核肌纤维显著减少，营养不良相关蛋白β-肌营养不良蛋白聚糖（β-DG）和α-肌聚糖（α-SG）以及神经元一氧化氮合酶（nNOS）恢复。被注射的dko小鼠还显示出体重增加、运动性能改善和寿命延长。使用HDAdv，我们甚至可以通过将治疗基因转移到多个骨骼肌中来治疗DMD模型小鼠。我们的研究结果表明，多次肌内注射携带全长肌营养不良蛋白cDNA的HDAdv可以减轻DMD患者的症状并补偿功能丧失。
Duchenne muscular dystrophy (DMD) is a fatal, progressive, muscle-wasting disease caused by defects in the dystrophin. No viral vector except the helper-dependent adenovirus vector ( HDAdv) can package 14-kilobase (kb) full-length dystrophin complementary DNA ( cDNA), and HDAdv is considerably safer than old-generation adenovirus vectors because of the large-size deletion in its genome. We have generated HDAdv that carries myc-tagged murine full-length dystrophin cDNA ( HDAdv-myc-mFLdys). We injected it into multiple proximal muscles of 7-day-old utrophin/dystrophin double knockout mice (dko mice) ( which typically show symptoms quite similar to human DMD) because the proximal muscles are affected in DMD patients. Eight weeks after the injections, the transduced dystrophin was widely expressed, and we found a significant reduction in centrally nucleated myofibers and the restoration of the dys-trophin-associated proteins, beta-dystroglycan ( beta-DG) and alpha-sarcoglycan (alpha-SG), as well as neuronal nitric oxide synthase ( nNOS). The injected dko mice also showed an increase in body weight, an improvement in motor performance, and a prolongation of life span. Using HDAdv, we could treat DMD model mice even by transferring the therapeutic gene into multiple skeletal muscles. Our results suggest that multiple intramuscular administrations of HDAdv carrying full-length dystrophin cDNA may reduce symptoms and compensate for lost functions in DMD patients.