Ex vivo gene transfer to mature skeletal muscle by using adenovirus helper cells.
Ex vivo gene transfer to mature skeletal muscle by using adenovirus helper cells.
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使用腺病毒辅助细胞将基因离体转移至成熟骨骼肌。
DOI：
10.1002/jgm.480
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发表时间：
2004
期刊：
影响因子：
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通讯作者：
Suzuki,Satoru
中科院分区：
文献类型：
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作者：
Kimura,Shigemi;Ikezawa,Makoto;Cao,Baohong;Kanda,Yasunari;Pruchnic,Ryan;Cummins,James;Huard,Johnny;Miike,Teruhisa;Suzuki,Satoru
背景腺病毒基因转移到成人骨骼肌受到几个主要限制，包括宿主免疫反应和成熟依赖的肌纤维感染力的丧失。在克服成熟依赖的腺病毒转导方面，经活体注射比直接注射病毒更有效。在这里，我们研究了利用辅助细胞来提高Fex vivogene转移到成年小鼠骨骼肌的效率。方法用mdx鼠的原代成肌细胞培养携带5型腺病毒E1基因的新的产生细胞(E32细胞)。将携带LacZ基因的E1缺失的第一代腺病毒感染E32细胞和293细胞。结果注射5d后，成年小鼠骨骼肌中可检测到LacZ阳性成熟肌纤维。293细胞和E32细胞的基因转导效率分别是成肌细胞介导的基因转导效率的6.2倍和3.6倍。结论293和E32辅助细胞转导的腺病毒基因转导效率高于成肌细胞基因转导和病毒直接注射。这些辅助细胞也能将腺病毒基因转移到成熟的肌纤维中。这种方法促进腺病毒基因转移到成熟肌纤维的机制尚不清楚；然而，我们假设，在转导的293和E32辅助细胞群中，细胞病变效应(CPE)的活体发生促进了腺病毒对肌纤维的转导。版权所有©2004 John Wiley&Sons，Ltd.
BackgroundAdenoviral gene transfer to adult skeletal muscle is hindered by several major limitations, including host immune responses and maturation‐dependent loss of myofiber infectivity.Ex vivogene delivery is more efficient than direct viral injection in surmounting maturation‐dependent adenoviral transduction. Here we investigated the use of helper cells to improve the efficiency ofex vivogene transfer to adult mouse skeletal muscle.MethodsNew producer cells carrying the E1 gene of adenovirus type 5 (E32 cells) were developed using primary myoblasts from mdx mice. The E32 cells and 293 cells were infected with an E1‐deleted first‐generation adenovirus carrying the LacZ gene. These transduced helper cells were injected into the skeletal muscle of adult mdx and SCID mice.ResultsLacZ‐positive mature myofibers were detected in the skeletal muscle of adult mice sacrificed 5 days post‐injection. The gene transfer efficiency using 293 cells and E32 cells was 6.2 and 3.6 times higher than myoblast‐mediated gene transfer, respectively.Ex vivogene transfer of these cell types led to a better outcome than did direct adenoviral injection.ConclusionsWe achieved more efficient adenoviral gene transduction by using 293 and E32 helper cells than by myoblast‐mediated gene transfer and direct viral injection. These helper cells also enabled adenoviral gene transfer to mature myofibers. The mechanisms by which this method facilitated adenoviral gene transfer to mature myofibers remains unclear; however, we hypothesize that thein vivooccurrence of cytopathic effects (CPE) in the transduced 293 and E32 helper cell populations facilitated the improved adenoviral transduction of myofibers. Copyright © 2004 John Wiley & Sons, Ltd.