1997 Fiscal Year Final Research Report Summary
Development of animal models with hepatitis C and hepatocellular carcinoma by transgenic techniques
| Project/Area Number |
08457077
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | Nihon University |
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Principal Investigator |
ESUMI Mariko Nihon University, School of Medicine Associate Professor, 医学部, 助教授 (30167291)
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| Co-Investigator(Kenkyū-buntansha) |
KATO Tamami Nihon University School of Medicine, Research Associate, 医学部, 助手 (70224516)
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| Project Period (FY) |
1996 – 1997
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| Keywords | Hepatitis C virus / Hepatocellular carcinoma / Traansgenic mouse / Methylation / 5-Azacytidine |
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| Research Abstract |
1. Production of transgenic mice
To establish an animal model of hepatitis C and hepatocellular carcinoma, we generated 31 transgenic mice carrying hepatitis C virus (HCV) genome : 3 lines of transgenic mice carrying a partial genome including the structural protein region under the control of the albumin promoter/enhancer (Alb-HN3.8), 19 lines with the whole genome of HCV (Alb-HN2) and 9 lines with the whole genome under the control of the SRalpha promoter (ME-HN2). HCV-specific RNA was little expressed within 8 weeks after birth.
2. Induction of transgene expression
To analyze the mechanism of transgene inactivation, we examined methylation inactivation of the transgene. Methylation-sensitive restriction enzyme fragment analysis and mapping of methylated cytosine by bisulfite-genome sequencing showed that the transgene was extensively methylated. A demethylating agent, 5-azacytidine, induced HCV gene expression in 4 of 6 lines examined. These results suggest that methylation of HCV cDNA is a cause of its suppressive expression in transgenic mice.
3. Histopathological changes of transgenic liver
We investigated long-term pathological changes in liver of transgenic mice. Hepatitis such as semi-acute phase of fulminant hepatitis and liver atrophy with lymphocyte infiltration and ascitic accumulation was observed in mice with 7 to 22 months of age : I of 1 mouse in line Alb-HN3.8-l, 1 of 4 mice in line Alb-HN2-48, and 1 of 6 mice in line ME-HN2-5. The expression of HCV RNA was detected in these liver tissues. As for line Alb-HN2-44, dysplastic changes were observed in 2 of 17 transgenic livers. One of them developed adenoma. These results suggest that only the expression of transgene induces pathological change of liver. However, it is further necessarily examined whether these changes observed are transgene-associated.
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Research Products
(21 results)
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[Publications] Kato T., Ahmed M., Yamamoto T., Takahashi H., Oohara M., Ikeda T., Aida Y., Katsuki M., Arakawa Y., Shikata T., and Esumi M.: "Inactivation of hepatitis C virus cDNA transgene by hypermethylation in transgenic mice" Arch.Virol.141. 951-958 (1996)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Esumi M., Hayashi N., Takahashi H., Shikata T., E to T., Moriyama M., Arakawa Y., Eto T., Nishihara T., Nozaki C., and Mizuno K.: "Immunoreactive core peptides of hepatitis C virus produced in Escherichia coli and in vitro DNA amplification-restricted transcription-translation system." J.Virol.Methods. 59. 91-98 (1996)
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「研究成果報告書概要(欧文)」より
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[Publications] Cerino A., Bissolati M., Cividini A., Nicosia A., Esumi M., Hayashi N., Mizuno K., Slobbe R., Oudshoorn P., Silini E., Asti M., and Mondelli M.U.: "Antibody responses to the hepatitis C virus E2 protein : relationship to viraemia and prevalence in anti-HCV seronegative subjects." J.Med.Virol. 51. 1-5 (1997)
Description
「研究成果報告書概要(欧文)」より
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