| Project/Area Number |
09671797
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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 |
Ophthalmology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
TAKAHASHI Masayo Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (80252443)
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| Co-Investigator(Kenkyū-buntansha) |
MANDAI Michiko Kyoto University, Graduate School of Medicine, Assistant Professor, 医学研究科, 助手 (80263086)
HONDA Yoshihito Kyoto University, Graduate School of Medicine, Professor, 医学研究科, 教授 (90026930)
谷原 秀信 京都大学, 医学研究科, 講師 (60217148)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1997: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | PVR / NF_<kappa>B / E2F / gene transfer / NFkBデコイ / 網膜芽細胞腫 / 変異型Rb遺伝子導入 / 遺伝子治療 / NFkB転写因子 |
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| Research Abstract |
Purpose. Previous studies suggested that proliferation and migration of retinal pigment epithelial cells are deeply involved in pathogenesis of proliferative vitreo retinopathy (PVR). Recently it is reported that decoys, double-stranded phosphorothioate oligonucleotides, have the same sequence of binding site of transcription factors. In this study we tested the effect of E2F decoys and NFkappaB decoys in the proliferation of cultured human retinal pigment epithelial (RPE) cells and in vivo animal model of PVR.Methods. HVJ cationic liposomes were prepared by replacing phosphatidylserine of HVJ liposomes (Hangai, et al, 1996) to DC cholesterol. Results. It was shown that E2F decoys highly combined to nuclear extracts from human fibroblast cells and NFkappaB decoys also combined to nuclear extracts from human RPE cells stimulated by interleukin 1beta (IL-1beta). RT-PCR indicated that E2F decoys introduced into human fibroblast decreased expression of important factor that is regulating cell cycle, it showed that NFkappaB decoys introduced into human RPE cells reduced transcripts of IL-1beta in IL-1beta stimulated RPE cells. BrdU labeling index and DNA synthesis (H-thymidine uptake) clarified that cell proliferation was highly inhibited with E2F decoys. Cells on culture dishes with NFkappaB decoys were significantly inhibited cell migrations compared with scrambled decoys. In vivo experiment NFkappaB decoys, transferred by HVJ cationic liposomes into human cultured fibroblast, were injected in the vitreous cavity of rabbit PVR model. The appearances of rabbit PVR were scored by a classification of Blumenkraz et al. The incidences of PVR were not significantly different between with NFkappaB decoys and with scrambled decoys. Conclusions. These results suggest that E2F decoys and NFkappaB decoys have the potential to treat proliferative vitreoretinopathy in vitro and it is needed more study to vitro use.
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