Ex vivo gene transfer using adenovirus-mediated full-length dystrophin delivery to dystrophic muscles
Ex vivo gene transfer using adenovirus-mediated full-length dystrophin delivery to dystrophic muscles
复制标题
DOI：
10.1038/sj.gt.3300549
复制
发表时间：
1998-01-01
期刊：
影响因子：
5.1
通讯作者：
Huard, J
中科院分区：
文献类型：
--
作者：
Floyd, SS;Clemens, PR;Huard, J
杜氏肌营养不良症（DMD）是一种X连锁隐性肌肉疾病，其特征在于缺乏肌营养不良蛋白表达。成肌细胞移植和基因治疗具有恢复肌营养不良蛋白的潜力，从而减少与疾病相关的肌无力。在这项研究中，我们提出的数据成肌细胞介导的全长肌营养不良蛋白的mdx（肌营养不良蛋白缺陷）小鼠肌肉作为自体成肌细胞转移的模型离体基因转移。用腺病毒载体转导同基因原代mdx成肌细胞和永生化mdx细胞系，所述腺病毒载体具有缺失的所有病毒编码序列并编码β-半乳糖苷酶和全长肌营养不良蛋白。随后，将这些转导的成肌细胞注射到营养不良的最大肌肉中，在那里注射的细胞恢复肌营养不良蛋白以及肌营养不良蛋白相关蛋白。从过表达肌营养不良蛋白的转基因小鼠中分离的mdx成肌细胞移植后，mdx肌肉中发生了更大量的肌营养不良蛋白替代，这表明工程化自体成肌细胞表达大量肌营养不良蛋白可能是有益的。离体方法具有使其可用于基因转移至骨骼肌的属性，包括：（1）产生能够再生肌营养不良蛋白并将其恢复至营养不良肌肉的成肌细胞库;和（2）与腺病毒介导的直接基因递送相比，实现更高水平的基因转移至营养不良肌肉。然而，如在直接基因转移研究中所观察到的，离体方法也引发细胞免疫应答，这限制了转基因表达的持续时间。
Duchenne muscular dystrophy (DMD) is an X-linked recessive muscle disease characterized by a lack of dystrophin expression. Myoblast transplantation and gene therapy have the potential of restoring dystrophin, thus decreasing the muscle weakness associated with the disease. In this study we present data on the myoblast mediated ex vivo gene transfer of full-length dystrophin to mdx (dystrophin deficient) mouse muscle as a model for autologous myoblast transfer. Both isogenic primary mdx myoblasts and an immortalized mdx cell line were transduced with an adenoviral vector that has all viral coding sequences deleted and encodes beta-galactosidase and full-length dystrophin. Subsequently, these transduced myoblasts were injected into dystrophic max muscle, where the injected cells restored dystrophin, as well as dystrophin-associated proteins. A greater amount of dystrophin replacement occurred in mdx muscle following transplantation of mdx myoblasts isolated from a transgenic mouse overexpressing dystrophin suggesting that engineering autologous myoblasts to express high amounts of dystrophin might be beneficial. The ex vivo approach possesses attributes that make it useful for gene transfer to skeletal muscle including: (1) creating a reservoir of myoblasts capable of regenerating and restoring dystrophin to dystrophic muscle; and (2) achieving a higher level of gene transfer to dystrophic muscle compared with adenovirus-mediated direct gene delivery. However, as observed in direct gene transfer studies, the ex vivo approach also triggers a cellular immune response which limits the duration of transgene expression.