| Project/Area Number |
09670686
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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 |
Neurology
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| Research Institution | National Institute of Neuroscience, NCNP |
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Principal Investigator |
TAKEDA Shinichi Department of Molecular Genetics, National Institute of Neuroscience, NCNP, Section Chief, 神経研究所・遺伝子工学研究部, 室長 (90171644)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAMOTO Kanji Department of Molecular Genetics, National Institute of Neuroscience, NCNP, 神経センター・神経研究所・遺伝子工学研究部, 流動研究員
MIYAGOE Yuko Department of Molecular Genetics, National Institute of Neuroscience, NCNP, 神経センター・神経研究所・遺伝子工学研究部, 研究員
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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 |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1998: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | ceruloplasmin deficiency / ceruloplasmin / neuronal cell death / reactive oxygen / gene therapy / adenovirus vector / knock out mice / iron deposition / 活性化酵素 / 原発性セルロプラスミン欠損症 / アデノウイルスベク- |
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| Research Abstract |
Hereditary ceruloplasmin deficiency is an autosomal recessive disorder characterized by middle age onset of involuntary movement, cerebellar ataxia, dementia, and diabetes mellitus. Marked iron deposition is observed in liver, pancreas, and basal ganglia with neuronal cell death. We detected a point mutation in the ceruloplasmin gene on a family with hereditary ceruloplasmin deficiency. In this family, the mutaion causes an abnormal splicing in the ceruloplasmin mRNA, resulting in emergence of premature stop codon and truncation of the C-terminus of ceruloplasmin. It is possible that tissue iron deposition and neuronal cell death are involved in lack of ferroxidase activity of ceruloplasmin and its function as a free ragical scavenger. For the development of gene therapy of the disease, we have generated a recombinant adenovirus vector expressing ceruloplasmin cDNA.We have cloned the full-length ceruloplasmin cDNA (3.7 kb) by means of RT-PCR.Using COS-TPC method, we generated an adenovirus vector, incorporating the cDNA under the control of the CAG promoter. The ceruloplasmin expression was confirmed by the adenovirus vector infection into cultured fibroblasts (1OT1/2), COS cells, and myoblasts (C2), followed by Western-blot analysis and immunocytochemistry. On the other hand, we developed ceruloplasmin-deficient mice. We will perform in vivo gene transfer into the liver of ceruloplasmin-deficient mice and in vitro gene transfer into primary culture of astrocytes.
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