Changes in Expression of Micrornas in Rat Liver Following Bile Duct Ligation

To develop therapies for preventing irreversible liver damage, it is essential to understand the molecular mechanisms responsible for liver cirrhosis. The production of a large and comprehensive data base of the miRNA profiles of bile duct obstruction-associated genes will assist the elucidation, by bioinformatic techniques, of those miRNA that are likely to contribute to fibrinogenesis and of those genes that may be responsible for fibrinolysis. The objectives are to use microarrays for a high throughput quantitative analysis of temporal changes in miRNA expression profiles in a rat bile duct ligation model, as well as defining those miRNA profiles characteristic of fibrinogenesis and fibrinolysis. In order to achieve these objectives the following four Specific Aims will be pursued 1) To identify the candidate miRNA related to bile duct ligation. A large-scale screen of miRNA arrays will be used to define miRNRAs that are regulated in bile duct ligated rats. The common regulated miRNA of variable cirrhosis models will be the priority to be assessed. 2) To validate the regulation of the bile duct ligated miRNA and determine their cellular localization. The expression profiles of the miRNAs will be validated by quantitative RT-PCR analysis, and cellular localization established by in situ hybridization. 3) To establish bioinfomaticaaly, the potential target genes of regulated miRNA. The significant down regulated genes after bile duct ligation and will be compared to the candidate miRNA from website. 4) To test the capacity of fibrinogenesis or fibrinolysis of candidate miRNAs in vitro and in vivo. Those miRNAs that fulfill all the criteria of a candidate miRNA in terms of their expression profile, cellular localization, bioinformatically predicted, will be delivered into liver in vitro and in vivo and fibrinogenesis will be assessed.

Project ID:PC10108-0813
External Project ID:NSC101-2314-B182-021-MY3