| Project/Area Number |
07670243
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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 |
Experimental pathology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HIGUCHI Keiichi Kyoto University, Chest Disease Research Institute, Lecturer, 胸部疾患研究所, 講師 (20173156)
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| Co-Investigator(Kenkyū-buntansha) |
HOSOKAWA Masanori Kyoto University, Chest Disease Research Institute, Associate Professor, 胸部疾患研究所, 助教授 (00127135)
國貞 隆弘 京都大学, 医学部, 講師 (30205108)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1996: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1995: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | Gene therapy / amyloidosis / adenovirus / mouse / CAG promoter / AApoAII / apoA-II / amyloid fibrilization / アデノウィルス / CAGプロモーター / コンジェニックマウス / apoA-II蛋白 / 老化促進モデルマウス |
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| Research Abstract |
Since, the effective treatment to amyloidosis is only the transplantation of the liver, we have done the basic studies for developing the gene therapeutic approach to amyloidosis. We used newly made congenic mice, R1.P1-Apoa2^c which are model mice of severe senile amyloidosis.
1.Genetic analysis of mouse senile amyloidosis : In the congenic mice, R1.P1-Apoa2^<b/c> which are heterozygous for wild type B and amyloidogenic type C apoA-II,amyloid deposition was significantly slight. This finding suggested the possible availability of gene therapeutic approach. 2.Primary culture of the hepatic cell of the amyloidogenic mouse strain : Hepatic cells were isolated and cultured from R1.P1-Apoa2^c mice. 3.Evaluation system for the effectiveness of the gene therapy : Since the onset of amyloid deposition is usually after 8-12 month of age, we tried to develop a new system for evaluation the effect of gene therapy in relatively short period. Intravenous injection of amyloid fibrils AApoAII in R1.P1-Apoa2^c induced amyloid deposition at one month after injection. In R1.P1-Apoa2^<b/c>, amyloid deposition was significantly inhibited. 4.Adeno-virus with wild type apoA-II cDNA : Cosmid cassette (pAdexCAwt) which has human type 5 adenovirus (Ad5) and strong promoter unit, CAG (cytomegarovirus enhancer, chicken-actin promoter rabbit-globin polyA) was kindly provided by Dr.I.Saito in Tokyo University. Wild type apoA-II cDNA (Apoa2^c) was subcloned into Swal site of the cosmid and co-transformed with parental adeno virus DNA-TCP into the 293 cell. Several adeno viruses with wild type apoA-II cDNA produced by homologous recombination was selected.
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