| Project/Area Number |
16591619
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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 | Kitasato University |
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Principal Investigator |
SATOH Takefumi Kitasato University, Medicine, Assistant Professor, 医学部, 講師 (50286332)
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| Co-Investigator(Kenkyū-buntansha) |
IRIE Akira Kitasato University, Medicine, Assistant Professor, 医学部, 講師 (50193694)
IWAMURA Masatsugu Kitasato University, Medicine, Assistant Professor, 医学部, 講師 (20176564)
FUJITA Tetsuo Kitasato University, Medicine, 医学部, 助手 (00306599)
BABA Shiro Kitasato University, Medicine, Professor, 医学部, 教授 (00051889)
田畑 健一 北里大学, 医学部, 助手 (20327414)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 2005: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2004: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | Gene therapy / Interleukin-12 / Renal cell carcinorma / 国際研究者交流 / 米国 |
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| Research Abstract |
We have documented previously that adenovirus-mediated IL-12 gene therapy is effective for orthotopic tumor control and suppression of pre-established metastases in a preclinical prostate cancer model. The observation of multiple immunocyte activities that potentially could develop into a systemic antitumor immune response involving the generation of memory T cells was evident, and the results of an analysis of distant antimetastatic activity in response to local injection of AdmIL-12 further supported this. To further explore this, we evaluated efficacy of AdmIL in situ gene therapy for preclinical renal cell carcinoma.
We have evaluated CAR expression in urinary tumor cell lines, because renal cell carcinoma is notorious for less expressing the CAR. However, the expression level of CAR (CAR/beta-action ratio) in Renca cells is same as DU145 and infection efficacy of AdLac-Z in Renca cells was 40%. According to these results, AdLac-Z can infect in Renca cells and Ad5 may useful for RCC model.
Based on these results, Renca cells were infected with increasing dose of adenoviral vectors containing mIL-12 (AdmIL-12, 0-400 PFU/cell) or〓-gal (Adv/CMV/〓-gal) in vitro to determine cytotoxicity of AdmIL-12 transfection, and to mesure the level of IL-12 production by capture ELISA. At 24 and 48 h following viral infection, there was no significant differences in cell number per well at any MOI compared to control wells (MOI of 0). Transfection of with increasing numbers of AdmIL-12 PFU resulted in a dose-dependent increase in IL-12 secreted into the medium.
For orthotopic tumor inoculation, BALB/c mice were anesthetized with sodium pentobarbital. Injection of 5000 cells (Renca) in 10 〓l directly into the right subcapsule of the kidney resulted in efficient and producible orthotopic tumor formation. We are still investigating to establish pre-clinical RCC animal model for this study. Hopefully, this therapeutic approach will be useful for specific clinical applications.
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