2002 Fiscal Year Final Research Report Summary
High efficiency gene transfer to skeletal muscle inadult mouse for Duchenne muscular dystrophy using adenovirus helper cells.
| Project/Area Number |
12670761
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pediatrics
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| Research Institution | Kumamoto University |
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Principal Investigator |
KIMURA Shigemi Kumamoto University, Child Development, Assistant professor, 医学部附属病院, 助手 (60284767)
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| Project Period (FY) |
2000 – 2002
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| Keywords | adult skeletal muscle / adenovirus / helper cells / 293 cells / CPE / Duchenne muscular dystrophy |
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| Research Abstract |
We have developed a strategy for achieving an efficient gene transduction of E1-deleted adenovirus to skeletal muscle in adult mice (>2 months old). New producer cells carrying the E1 gene of adenovirus type 5 (E32 cell) were developed using primary myoblasts from mdx mouse. The new helper cells and 293 cells were infected with E1-deleted adenovirus carrying the gene-encoding lacZ and were injected into skeletal muscle of adult mdx mice. Many lacZ-positive fibers were detected in the skeletal muscle of adult mdx mice, which were sacrificed at 5 days post-injectiorL This new method was compared with regular myoblast-mediated gene transfer and direct injection of adenovirus. The efficiency of gene transduction, using 293 cells and E32 cells, were 6.2 times and 3.6 times as high as myoblast-mediated gene transfer, respectively. We hypothesize that the adenovirus-infected 293 cells release some type of factor that breaks down the mature myofibers' barrier to viral infection, thereby increasing transduction. In conclusion, the efficiencies of gene transduction using 293 cells and E32 cells were higher than myoblast-mediated gene transfer and direct viral injection. More importantly, those helper cells enabled efficient gene transduction of adult skeletal muscle.
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