1997 Fiscal Year Final Research Report Summary
In vivo gene transfer methods into bladder without viral vectors
| Project/Area Number |
08671833
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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 |
Urology
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| Research Institution | OSAKA CITY UNIVERSITY |
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Principal Investigator |
KISHIMOTO Taketoshi Osaka City Univ.Med.School, (Dept.Urology) Professor, 医学部, 教授 (00047078)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Seiji Osaka City Univ.Med.School, (Dept. Urology), Assistant Professor, 医学部, 講師 (20158695)
SUGIMURA Kazunobu Osaka CIty Univ.Med.School, (Dept.Uroligy), Assistant Professor, 医学部, 講師 (90187659)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Gene transfer / Bladder / HVJ-liposome / Particle gun / Electroporation |
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| Research Abstract |
objective GEne therapy has been adapted as a new strategy for treating cancers. When considering this application to bladder cancer, safe, efficient and repeatable in vivo geno transfer metods are essential. Here, we examined three in vivo gene transfer methods without viral vectors.
Materials and methods Hemagglutinating virus of Japan (HVJ)-liposomes possessing the membrane fusion activity were intraluminally injected into rat bladders. Using a particle gun, rabbit bladder mucosa was bombarded with DNA-Coated gold microcarriers. Electrotransfection was also examined in rabbit bladder by pulse direct currents (0.15-0.2 A, 50msec, repeated 8 times) generated between needle electrodes after submucous injection of DNA solution. beta-galactosidase gene and chloramphenicol acetyl-transferase (CAT) gene were used as marker genes.
Results HVJ-liposomes efficiently transfected superficial layrs of urothelium with a peak of expression on day 5. The particle gun produced non-uniform but efficient transfection in deeper layrs of the urothelim. By electransfection, submucous interstitial cells were transfected as well as urothelim. No major complications were observed in these three metheds.
Conclusion HVJ-liposomes are potentially useful for treating carcinoma in situ. With further refinement, the latter two methods may be suitable for adjuvant therapies in treating localized bladder tumors.
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