I型干扰素依赖的病毒刺激诱导的长链非编码RNA在抗病毒天然免疫应答中的功能及机制研究

项目名称： I型干扰素依赖的病毒刺激诱导的长链非编码RNA在抗病毒天然免疫应答中的功能及机制研究

项目编号： No.31470871

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 王品

作者单位： 中国人民解放军第二军医大学

项目金额： 85万元

中文摘要： 机体抗病毒天然免疫应答一直以来是免疫学研究的重点，然而过去的研究都集中在编码蛋白的基因，目前还没有lncRNA在这方面的研究报道。前期实验我们用lncRNA表达芯片在小鼠巨噬细胞中筛选到了一批能够在病毒刺激后依赖IFN-I信号被诱导表达的lncRNA。用siRNA进行功能筛查，发现其中lincRNA-203和lincRNA-220能够调控IFN-I的产生和病毒的复制。后续将进一步用高表达和干扰稳筛细胞系、基因操作小鼠进行体外和体内多种病毒刺激后检测分析，深入研究该lncRNA在抗病毒天然免疫应答中的功能，同时检测其在人类细胞中同源基因的功能。在机制上我们将筛查其调控的信号通路转录因子、表达芯片检测其调控的基因、质谱筛查其发挥作用的结合蛋白，并探索该lncRNA在病毒刺激后表达上调的分子机制。该研究将确定lncRNA在天然抗病毒免疫中的作用和机制，为临床病毒感染和IFN相关治疗提供研究基础。

中文关键词： 免疫反应；长链非编码RNA；抗病毒免疫；细胞信号传导；I型干扰素

英文摘要： The regulation of antiviral innate immune response has long been intensively explored. However, almost all of these studies are focused on the role of coidng genes while ignoring the large majority of noncoding genes,especially for long noncoding RNA.With microarray, we have explored the expression profile of lncRNA and identified a cluster of lncRNAs induced in virus infection through a IFN-I-signaling dependent manner. After function scanning with their siRNA, we found lincRNA-203 and lincRNA-220 could regulate IFN-I produciton and virus replication. This project intend to further confirm the role of these two lncRNA in vitro using stably transfected cell line and in vivo using gene-modified mice in the model of different type virus infection, SeV,VSV, HSV.Then we will investigate signaling pathways and genes regulated by these lncRNAs through immunoblot and microarray detection, and identify their interacting proteins which mediate their function through mass spectrometry and colocalization analysis. We will also explore the underlying mechanism of these lncRNAs upregualtion after virus infection through promotor analysis, ChIP,and gene reporter assay. This study will clarify the noval role of lncRNA in innate immune response and provide potential targets or markers for the therapy of clinical virus infection disease or IFN treatment.

英文关键词： immune response;long noncoding RNA;antiviral immunity;intracellular signaling;type I IFN