项目名称: miR-491通过调控T细胞的增殖和凋亡在诱导T细胞衰竭中的作用机制研究
项目编号: No.81202310
项目类型: 青年科学基金项目
立项/批准年度: 2013
项目学科: 医学免疫学、法医学
项目作者: 肖斌
作者单位: 中国人民解放军第三军医大学
项目金额: 23万元
中文摘要: T细胞衰竭是指T细胞在特定情况下出现的功能抑制或失衡,其发生机制仍不清楚。研究证实,miRNAs可参与调控T细胞的增殖、凋亡和分化,那么miRNAs是否也能调控T细胞衰竭?我们在前期研究证实,miR-491在衰竭T细胞中表达上调,过表达miR-491能抑制T细胞增殖,促进T细胞凋亡,且miR-491启动子序列中可能含有SMAD结合位点,提示miR-491可能参与诱导T细胞衰竭。本项目拟利用逆转录病毒和AntagomiR系统,体内和体外过表达及抑制miR-491,研究其对T细胞增殖、凋亡以及衰竭表型的影响,同时通过荧光素酶、siRNA、CHIP等鉴定miR-491的靶基因和上游表达调控因子,以验证假说:miR-491可能通过下调靶基因(如Bcl-w、TCF7和CDK4)的表达,抑制T细胞增殖,促进T细胞凋亡,从而诱导T细胞衰竭,而miR-491的高表达可能受TGF-β-SMAD通路的调控。
中文关键词: miR-491;T细胞衰竭;增殖;凋亡;肿瘤
英文摘要: T cell exhaustion is a state of T cell dysfunction that arises during chronic infections and cancer, the exact mechanism is not clear. MicroRNAs are small non-coding RNAs, which can negatively regulate gene expression. There are increasing evidences that microRNAs can regulate the function of T cells including proliferation, apoptosis, and differentiation. Whether microRNAs are involved in the regulation of T cell exhaustion? In our previous study, we found miR-491 was significantly up-regulated in exhausted T cells, and overexpression of miR-491 can inhibit the proliferation and induce apoptosis of T cells. Above data suggest miR-491 may play potential role in the induction of T cell exhaustion. In current project, we plan to study the role of miR-491 on the proliferation, apoptosis and exhaustion phenotype in vitro and in vivo by using retrovirus expression system and antagomiR which can overexpress or inhibit miR-491 level. Then we want to identify the target gene and transcription factor of miR-491 by using luciferase report system, siRNA, western-blot, and Chip. In conclusion, our hypothesis is that miR-491 is up-regulated in exhausted T cells, and miR-491 may inhibit the proliferation and induce the apoptosis of T cells by down-regulating the target genes of Bcl-w, TCF-7, and CDK-4, it will result in the
英文关键词: miR-491;T cell exhaustion;proliferation;apoptosis;tumor