| Project/Area Number |
11671764
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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 | Kansai Medical University |
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Principal Investigator |
YAMAMOTO Chikako Kansai Medical University School of Medicine, Assistant, 医学部, 助手 (80288844)
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| Co-Investigator(Kenkyū-buntansha) |
KANEDA Yasufumi Osaka University, School of Medicine, Professor, 医学部, 教授 (10177537)
OGATA Nahoko Kansai Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (60204062)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | choroidal neovascularization / transcriptional factor / decoy / HVJ liposome / gene transfection |
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| Research Abstract |
(1) Expression of transcriptional factors in experimental choroidal neovascularization.
Intense laser burns were applied to the posterior retina of pigmented rats to induce choroidal neovascularization (CNV). Following administration, the eyes were removed and fixed, and then frozen and cut into thin sections. Immunohistochemistrical examination was done to these sections by anti-transcriptional factors (NFkB, Ets-1, HIF-1). In result, the expression of NFkB and Ets-1 were seen on early stage of CNV developing.
(2) Transfection of cis element "decoy" against transcriptional factor bindng site into choroidal neovascularization by HVj liposome method.
HVJ loposomes including fluorescein isothiocyanate (FITC) labeled couble stranded phosphorothioate oligodeoxynucleotides (S-ODNs) "decoy" against NFkB binding site (NFkB "decoy" S-ODNs) were prepared. Intense laser burns were applied to the posterior retina of pigmented rats to induce CNV.Immediately after photocoagulation, IVJ liposome suspension was injected into the vitreous or subretina. Following administration, fluorescein angiography (FAG) was performed and the eye s were removed. Frozen sections were observed fluorescence of FITC under a confocal scanning lase microscope. Paraffin sections were observed to evaluate the development of CNV, histopathologically. Eye administrated random controlled double-stranded S-ODNs as sontrals.
FAG showed hyperfluorescence at laser lesion in NFkB decoy S-ODNs injected group. However the leakage of dye was seaker in decoy S-ODNs injected group than that was observed in controls. FITC labeling was seen in CNV lesion. Histopathologically, the development of CNV was suppressed in NFkB "decoy" S-ODNs transfected eyes at early stage.
Inconclusion, we demonstrated that NFkB decoy S-ODNs were transferred into the choroidal neovascularization by HVJ liposome method. NFkB decoy S-ODNs would be an effective therapeutic agent for choroidal neovascularization treatment in clinical field.
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