1995 Fiscal Year Final Research Report Summary
Basic Research for Ex Vivo Gene Tharapy using Gene Targeting Method
| Project/Area Number |
06670771
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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 |
Pediatrics
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| Research Institution | The University of Tokyo |
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Principal Investigator |
NARITAKA Shinichi The University of Tokyo, Dept.of Pediatrics, 医学部・附属病院, 助手 (60179314)
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| Co-Investigator(Kenkyū-buntansha) |
HATA Kensaku The University of Tokyo, Dept.of Pediatrics, (Sanraku Hospital), 医学部・附属病院, 科長 (70189546)
HAYASHI Yasuhide The University of Tokyo, Dept.of Pediatrics, 医学部, 講師 (30238133)
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| Project Period (FY) |
1994 – 1995
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| Keywords | gene targeting / ex vivo gene therapy / in vivo gene therapy / Wilson disease / LEC rats / primary culture of hepatocytes / recombinant adenovirus vector / Lac Z gene of E.coli |
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| Research Abstract |
The aim of this study was to establish the method of ex vivo gene therapy.
wilson's disase is a genetic disease and this disease gene-copper transporting ATPase (ATP-7B) has been cloning recently.Long Evans Cinnamon (LEC) rat is the model of Wilson's disease.Genetically, the disease gene (Atp-7b) of LEC rat is 70% homology of Wilson's disease.Successful gene therapy of LEC rat is the beginning of gene correction of Wilson's disease.
We performed partial hepatectomy and the isolation of hepatocytes using Wister rats.Partial hepatectomy was successful, if we avoided bleeding from resected region.The isolation and primary cuture of hepatocytes was complicated and needed for special technique such as very careful handling and optical culture condition.
The LacZ gene of E.coli was inserted into normal hepatocytes by using recombinant adenovirus as vector.The expression of this gene was confirmed on the cultured hepatocytes.However, in vivo introduction of this gene into rats was not successful.This phenomenon suggested that either technical problem or essential problem like inserted gene or vector itself have related to the failure.Several problems still remains unknown.We needed to resolve these problems in future.
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