调节肝细胞胰岛素敏感性的新关键基因的筛选和研究

项目名称： 调节肝细胞胰岛素敏感性的新关键基因的筛选和研究

项目编号： No.31200591

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 生物物理、生化与生物分子学、生物力学与组织工程

项目作者： 张芳

作者单位： 中国科学院上海生命科学研究院

项目金额： 25万元

中文摘要： 2型糖尿病是一种复杂的慢性代谢疾病，胰岛素敏感性下降是其主要特征。目前改善胰岛素敏感性的方法已有较多研究，但在临床应用方面还存在不足。因此迫切需要寻找新的可调节胰岛素敏感性的关键基因。高通量测序技术具有多方面优势，适合全面筛选差异基因。本项目拟利用高通量测序技术对2型糖尿病和正常小鼠肝脏组织的基因表达谱进行定量检测；并通过生物信息学方法分析有显著差异的基因、信号通路和基因网络，结合文献资料，从中预测和筛选30-45个新的调节肝细胞胰岛素敏感性的候选基因；接着在细胞水平通过siRNA下调候选基因进行筛选，并用过表达方法进一步确定候选基因的有效性，筛选出至少2个可调节肝细胞胰岛素敏感性的候选基因；进而在动物水平利用尾静脉腺注射病毒和基因敲除小鼠等方法希望筛选出至少1个可以显著调节胰岛素敏感性的新关键基因；并对其作用的分子机制进行初步探索。本项目的顺利实施将为2型糖尿病的防治提供新的思路和线索。

中文关键词： 2型糖尿病；胰岛素敏感性；高通量测序；节律紊乱；SIRT1

英文摘要： Type 2 diabetes, as one of the most complex metabolic diseases, is characterized by the reduction of insulin sensitivity. Currently, there are many reseaerch on insulin sensitivity improvement, however, the escalating prevalence of type 2 diabetes and the limitations of currently available preventative and therapeutic options highlight the need for a more complete understanding of the pathogenesis of type 2 diabetes and searching for new key genes for insulin sensitivity regulation. Recently, high-throughput sequencing shows major advances in robustness, resolution and reproducibility based on its relatively unbiased and direct digital readout with sequencing by synthesis method, and is sutable for screening differentially expressed genes globally. Our project is to globally profile the gene expression of type 2 diabetic and normal mice liver with digital high-throughput sequencing, and predict 30-45 new candidate genes for hepatic insulin sensitivity regulation through bioinformation analysis of differentially expressed genes, pathways and networks. Next, screening at least 2 candidate genes at cell level, through downregulating and upregulating candidate genes with siRNA and overexpression step by step. Third, we plan to detect the effect of at least 1 candidate gene for insulin sensitivity improvement in vivo

英文关键词： Type 2 diabetes；Insulin sensitivity；High-throughput sequencing；Circadian misalignment；SIRT1