新疆维吾尔族2型糖尿病circRNA差异性表达及其调控网络机制研究

Uygurs are high-risk population with type 2 diabetes (T2DM) in Xinjiang, and their prevalence is significantly higher than other ethnic minorities in the same region. Recent studies suggest that the expression or functional abnormality of circRNA is closely related to the development of human diseases. However, the role of circRNA in T2DM has not been elucidated yet. To investigate the role of circRNA in the pathogenesis of Uygur T2DM in Xinjiang. This project intends to construct circRNA-miRNAs by using techniques of circRNA, mRNA, siRNA, lentivirus overexpression, fluorescence in situ hybridization, RNA-binding protein immunoprecipitation, luciferase reporter assay in three aspects of cell, animal and human. We construct the regulatory network of circRNA-miRNA-mRNA and identify the T2DM-related circRNAs and genes in Uygur, and explore its possible mechanism, and clarify the relationship between circRNA and its regulatory networks and the occurrence and development of T2DM in Xinjiang Uygur. To reveal the incidence of T2DM in Uygur Mechanisms and individualized therapeutic molecular targets provide the basis for screening.

新疆地区维吾尔族是2型糖尿病(T2DM)高发民族，其患病率明显高于同地区其他少数民族。新近研究提示，环状RNA(circRNA)的表达或功能异常与人类疾病的发生发展密切相关。然而circRNA在T2DM中的作用尚未阐明。为探讨circRNA在新疆维吾尔族T2DM发病机制中的作用。本课题拟在细胞、动物与人群三个层面，采用circRNA芯片、mRNA芯片、siRNA、慢病毒过表达、荧光原位杂交、RNA结合蛋白免疫沉淀、荧光素酶报告基因检测等技术，构建circRNA-miRNA-mRNA调控网络，鉴定维吾尔族T2DM相关的circRNAs和基因，并研究其可能作用机制，阐明circRNA及其调控网络与新疆维吾尔族T2DM发生发展的关系，为揭示新疆维吾尔族T2DM的发病机制及个体化治疗分子靶点的筛选提供依据。

环状RNA(circRNA)作为一种新的非编码RNA分子是表观遗传机制的重要组成部分，它可以通过吸附特定的微小RNA调控靶基因表达，参与糖尿病发病。本课题对比维吾尔族T2DM患者和糖耐量正常对照者circRNA、mRNA表达谱的差异，筛选出可能作为T2DM生物标志物的circRNA；应用生物信息学方法预测circRNA上高匹配值的miRNA结合位点、miRNA靶基因并注释差异表达的mRNA的功能及所涉及的信号通路，挑选出与糖尿病相关的mRNA，构建circRNA-miRNA-mRNA调控网络。研究circRNA通过miRNA对靶mRNA表达的调控作用，探讨cirRNA在T2DM发病机制中的作用，为深入研究circRNA在维吾尔族T2DM发病过程中的调控功能提供理论基础和实验证据。研究发现：1）hsa_circ_0042817、hsa_circ_0006532、hsa_circ_0004131在T2DM患者中高表达，hsa_circ_0042817在维吾尔族T2DM患者与汉族T2DM患者表达不同，具有成为维吾尔族T2DM分子生物标志物的潜在价值；2）T2DM患者CHIT1基因表达上调，CHIT1的高表达与糖脂代谢、胰岛功能密切相关；hsa_circ_0058213表达上调可能通过靶向结合hsa-miR-760调控CHIT1基因的表达，参与维吾尔族T2DM的病理生理过程；3）circ_ZFYVE1通过促进MIN6胰岛β细胞增殖、减少其凋亡，以及促进胰岛素转录因子PDX1表达，从而参与胰岛β细胞的功能调控；4）circRNA_0003340在T2DM大鼠不同病程上存在时间差异性表达，与T2DM病程中胰岛β细胞功能密切相关。