| Project/Area Number |
14580752
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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 |
Neurochemistry/Neuropharmacology
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| Research Institution | Fujita Health University |
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Principal Investigator |
NAKASHIMA Akira Fujita Health University, School of Medicine, Associate Professor, 医学部, 助教授 (20180276)
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| Co-Investigator(Kenkyū-buntansha) |
OTA Akira Fujita Health University, School of Medicine, Professor, 医学部, 教授 (10247637)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | tyrosine hydroxylase / mutant / Parkinson's disease / gene therapy / feedback inhibition |
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| Research Abstract |
Several studies have suggested that the direct virus vector-mediated delivery of transgenes of tyrosine hydroxylase (TH) involved in atecholamine synthesis in vivo may be available for gene therapy of Parkinson's disease (PD). However, TH expressed in mammalian cells cannot produce efficiently L-dopa, because the catalytic activity of TH is inhibited by the end-products catecholamines accumulated in the cells. The purpose of our study is to produce TH with high capability of L-dopa synthesis in the mammalian cells for an effective gene therapy of PD. We produced the mutant enzymes of human TH type1 (hTH1) and then examined the characteristic of them. 1)The replacement of N-terminal residues 30-40 of hTH1 by neutral or negatively charged residues decreased the inhibitory effect of dopamine on the catalytic activity. 2)Especially, the replacement of Arg^<37> -Arg^<38> and Ser^<40> of hTH1 by negatively charged Glu or Asp gave an efficient production of DA in vitro. 3)The efficient production of DA by the mutant enzymes was detected in AtT-20 neuroendcrine cells. 4)Moreover, the mutant enzymes of hTH1 revealed a high stability in other mammalian cell lines in addition to AtT-20 cells. Collectively, our study provides useful information for the refinement of the gene-therapy approach to obtain increased production of DA in vivo.
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