1999 Fiscal Year Final Research Report Summary
A novel gene transfer method for prostate-specific and potent expression and its utilization in gene therapy
| Project/Area Number |
10470337
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | KYOTO UNIVERSITY |
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Principal Investigator |
KAKEHI Yoshiyuki Kyoto University, Graduate School of Medicine, Associate Professor, 医学研究科, 助教授 (20214273)
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| Co-Investigator(Kenkyū-buntansha) |
TERAI Akito Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 講師 (50243019)
TERACHI Toshiro Kyoto University, Graduate School of Medicine, Associate Professor, 医学研究科, 助教授 (50207487)
KAKIZUKA Akira Osaka Biosience Institute, The 4<@D1th@>D1 Department, Director, 第4研究部, 部長 (80204329)
MIZUTANI Yoichi Kyoto University, Graduate School of Medicine, Instructer, 医学研究科, 助手 (10243031)
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| Project Period (FY) |
1998 – 1999
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| Keywords | Prostate cancer / PSA / gene therapy / LNCaP cell / androgen responsiveness / TSTA system / GFP / adenovirus vector |
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| Research Abstract |
We showed that an approximately 5.3-kb promoter region of the prostate-specific antigen (PSA) gene can replicate the endogenous expression pattern in an androgen-sensitive human prostate cancer LNCaP cells. We then developed a novel two-step transcriptional activation system (TSTA system) in which the PSA promoter drives an artificial transcriptional activator, GAL4-VP16 fusion protein, and it in turn activates transgene expressions under the control of GAL4-responsive element. By using this system, transgene expressions can be greatly augmented while maintaining prostate-specific expression. We applied this system to drive an expanded polyglutamine (ex-poly-Q), a potent proapoptotic molecule, to induce apoptosis selectively in PSA-positive prostate cancer cells. In order to confirm this system in vivo, we tried to construct two adenovirus vectors carrying PSA-promoter plus GAL4-VP16 or GAL4-resposive element plus ex-poly Q. We have, however, not yet isolated the adenovirus in which PSA-promoter plus GAL4-VP16 is transfected.
To-establish an animal model for the assessment of gene therapy, we isolated LNCaP cells that showed stable expression of green fluorescence protein(GFP) gene. When we inoculated this LNCaP-GFP cells orthotopically into the prostate of SCID mice, pulmonary metastatic foci could be detected much earlier than the conventional method that ever reported (4 weeks vs. 12 weeks). This system thus is considered to be utilized as a tactile in vivo model for the assessment of anti-metastatic therapy.
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