拟人化hERG基因突变及敲除的糖尿病动物模型的建立

项目名称： 拟人化hERG基因突变及敲除的糖尿病动物模型的建立

项目编号： No.81471014

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： 杨金奎

作者单位： 首都医科大学

项目金额： 150万元

中文摘要： 真实模拟人类糖尿病动物模型是糖尿病发病机制及药物新靶点研究的必要条件。目前的自发性糖尿病动物模型与人类的单基因及多基因遗传糖尿病存在明显差异。前期研究我们发现，hERG基因的错义突变是一个特殊大型家系发生糖尿病的原因；而hERG钾通道与KATP钾通道对胰岛β细胞释放胰岛素发挥互补性调节作用；阻断hERG可促进胰岛素分泌。本课题将构建拟人化hERG基因突变糖尿病动物模型。分别应用CRISPR-Cas9和TALEN基因敲除/修饰技术在大鼠和斑马鱼两种动物中构建hERG基因敲除和点突变靶向修饰糖尿病模型。用构建的大鼠基因敲除和靶向修饰模型研究hERG钾通道对β细胞释放胰岛素的影响及其作用机制。用构建的斑马鱼基因敲除模型研究hERG基因缺乏后对胰岛发育的影响及未来用于高通量筛选针对hERG钾通道的药物。该模型的构建将有助于深入了解人类胰岛素分泌的机制与糖尿病的发生，探索糖尿病新的治疗方法。

中文关键词： hERG；基因突变；胰岛β细胞；胰岛素分泌

英文摘要： Anthropomorphic animal models of diabetes are necessary for the research of pathogenesis and new pharmaceutical targets of diabetes. There are obvious differences between spontaneous diabetic animal models and human single gene or polygenic diabetes. Previously, we found that a missense mutation of hERG gene is the cause of a special diabetes in a large pedigree; hERG potassium channels and KATP potassium channels play a complementary role on pancreatic β cells release of insulin; blocking hERG can promote insulin secretion. This study will build anthropomorphic diabetic animal models with hERG gene mutation. We will apply CRISPR-Cas9 and TALEN knockout / modification technology to build hERG gene knockout and targeted modification/point mutations in the rats and zebrafish; use mice models of gene knockout and targeted modification in hERG gene to study its effects and its mechanisms on insulin release; use zebrafish models of gene knockout in hERG gene to study its mechanism on islet development and to be used for high- throughput screening of diabetic drugs. The model will help understanding of the mechanism of insulin secretion occurring in human diabetes, and exploring new treatment for diabetes.

英文关键词： hERG;gene mutation;islet beta cell;insulin secretion