供肝TCF7L2基因rs290487位点突变参与肝移植术后新发糖尿病发生的分子机制研究

项目名称： 供肝TCF7L2基因rs290487位点突变参与肝移植术后新发糖尿病发生的分子机制研究

项目编号： No.81470892

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 凌琪

作者单位： 浙江大学

项目金额： 73万元

中文摘要： 在青年基金支持下，申请者发现了供肝TCF7L2基因多态性与肝移植术后糖尿病（NODAT）之间的密切关联性并进行了分子机制的初探。围绕前期rs290487位点T→C突变引起14号外显子表达下降，肝细胞胰岛素敏感性和糖代谢能力减弱；免疫抑制剂引起TCF7L2表达下降这一重大发现，申请者拟通过TALEN技术、RQ-PCR array、ChIP-seq、小鼠条件性基因敲除和in silico分析等方法，开展以下研究：①TCF7L2基因rs290487位点突变对基因剪切调控、蛋白编码、转录因子绑定的影响研究；②第14号外显子在发挥TCF7L2蛋白功能中的作用（包括DNA结合、Wnt和PI3K等重要通路的激活等）及对肝糖代谢的影响研究；③免疫抑制状态对TCF7L2功能及肝糖代谢的作用研究。旨在阐明TCF7L2基因rs290487位点突变对肝糖代谢的调控作用及移植状态下引起NODAT的可能机制。

中文关键词： 肝移植；移植后糖尿病；基因多态性；剪切体

英文摘要： Under the support of National Natural Science Foundation of China (81100321), we found a close association between donor graft TCF7L2 gene polymorphisms and the risk of new-onset diabetes after transplantation (NODAT) and performed a primary exploration of the underline mechanism. The important findings were that T to C mutation at rs290487 locus could lead to a down-regulation of exon 14 expression, and a reduction of glucose uptake and insulin sensitivity in liver cells. We also found that high dose tacrolimus contributed to a decrease of glucose uptake of liver cells and reduce the expression of TCF7L2 protein. Based on the previous findings, we will use TALEN knock-in technique, RQ-PCR array, CHIP-seq, liver specific gene knock-out mice, and in silico analysis to perform the following study: ① the effect of TCF7L2 gene s290487 locus mutation on mRNA splicing, protein encoding, transcript factors binding and liver cell glucose uptake；② the role of exon 14 encoding in TCF7L2 protein function, including DNA binding and activation of Wnt and PI3K pathway, and hepatic glucose metabolism；③ the impact of immunosuppressant agent on TCF7L2 function and hepatic glucose metabolism. We aim to clarify the role of TCF7L2 gene rs290487 locus mutation in hepatic glucose metabolism, and the molecular mechanism of NODAT in liver transplantation.

英文关键词： liver transplantation;New-onset diabetes after transplantation;gene polymorphism;transcript variant