1991 Fiscal Year Final Research Report Summary
Trial of the expression vector DNA injection to the eye
| Project/Area Number |
02670796
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Ophthalmology
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| Research Institution | Juntendo University School of Medicine |
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Principal Investigator |
HOTTA Yoshihiro Juntendo University, Dept. of Ophthalmol., Assistant, 医学部・眼科, 助手 (90173608)
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| Co-Investigator(Kenkyū-buntansha) |
AKIYAMA Shuichi Juntendo University, Dept. of Ophthalmol., Assistant, 医学部・眼科, 助手 (10192915)
YOKOYAMA Toshiyuki Juntendo University, Dept. of Ophthalmol., Assistant, 医学部・眼科, 助手 (00191528)
FUJIKI Keiko Juntendo University, Dept. of Ophthalmol., Assist. Pro, 医学部・眼科, 講師
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| Project Period (FY) |
1990 – 1991
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| Keywords | vector / gene transfer / gene expression / gene therapy / vitreous / molecular biology / RNA |
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| Research Abstract |
Recent advance in molecular biology clarified the cause of the hereditary eye diseases which lead to blindness. A gene therapy for this disease may be possible if a cloned defective gene can be introduced into and expressed in the defective tissue. We studied the expression of defective gene after gene transfer in cultured cells and after the injection to the vitreous in the rabbit.
Structural and expression defects of the OAT (ornithine aminotransferase) gene have been demonstrated in gyrate atrophy which is an degenerative disease of the retina and choroid. We used OAT (-) Chinese hamster ovary (CHO) cells, which have negligible OAT activity, and fibroblasts from a gyrate atrophy patient (GA35 cell), which have negligible OAT mRNA and enzyme. Incorporation of vector pcDHOAT and synthesis of human OAT mRNAs and active enzyme were demonstrated in both cell types. The level of expression of human OAT was low in the GA35 cells in comparison to the CHO cells. Despite the limited. success, the ability to express active OAT in these OAT-deficient cells using an expression vector offers possibility of replacement gene therapy for gyrate atrophy.
We injected pcDHOAT DNA into the vitreous eavity in the rabbit. We isolated retinal tissue after three month later and extracted RNA. Unfortunately, northern blot analysis showed no human OAT mRNA in the rabbit retina. We will plan to use another vector and method of the injection. We also investigated Japanese patients with retinitis pigmentosa and Leber hereditary optic neuropathy using the molecular biological techniques, because these patients seem to be the candidate for the gene therapy.
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