与流感病毒致病相关的宿主长链非编码RNA的筛选鉴定及功能研究

It is now well known that long non-coding RNAs (lncRNAs) play important roles in multiple physiological and pathological progresses. As novel host factors, lncRNAs regulate the process of viral infection and replication, and are critically involved in host antiviral immunity. However, there is little information available about the precise function of lncRNAs in interaction between influenza virus and host. The precise mechanisms underlying involvement of lncRNAs in antiviral defense remain to be defined. In this research project, we use human and mouse lncRNA microarrays and RNA sequencing to examine the changes of lncRNA expression in host cells infected with or without influenza A virus. We will perform various assays using bioinformatics, cellular and molecular methods, and mouse models. The main aims of this project are discovery and identification of novel functional lncRNAs that play key roles in the pathogenesis of influenza A virus. Furthermore, we will determine mechanisms underlying their function during the pathogenesis of influenza A virus. Our previous studies identified a novel lncRNA named as lncRNA-up30 that was shown to be involved in infection and replication of influenza A virus. We will further investigate the precise roles and underlying mechanisms of lncRNA-up30 and other newly identified lncRNAs in pathogenesis of influenza A virus. In addition, using mouse models we will perform functional analysis of these lncRNAs in vivo. Taken together, this project will be focused on identification and characterization of novel functional lncRNAs that are critically involved in the pathogenesis of influenza A virus. These studies may provide a useful theoretical basis for development of novel antivirus drugs.

长链非编码RNA（lncRNAs）参与了机体的多种生理和病理过程，其作为新发现的一类宿主因子，在病毒致病过程中起重要调控作用。但迄今为止，有关lncRNAs在流感病毒与宿主互作中的作用及其机制仍不很清楚。本项目采用人源和鼠源lncRNA芯片与转录组分析，通过生物信息学研究，分析流感病毒感染前后宿主细胞内lncRNAs的表达谱，发现和鉴定新型与流感病毒致病过程密切相关的功能性lncRNAs，并对其作用机制进行深入研究。重点研究前期工作中筛选鉴定出的与流感病毒致病密切相关的新lncRNAs，通过体内外实验对lncRNA-up30等lncRNAs在流感病毒致病中的调控功能进行系统研究。本项研究旨在从分子、细胞和动物水平上阐释lncRNAs在宿主抗流感病毒感染中的作用及分子基础，为彻底阐明流感病毒的致病机理提供新的科学依据，为抗流感病毒药物的研发提供有价值的新靶标。

长链非编码RNA（lncRNAs）广泛存在于各类生命体内，种类繁多，数量巨大，在表观遗传调控、转录以及转录后调控等多种水平上对生命活动起着十分重要的作用。虽然近年来有关lncRNAs的研究在国际上越来越受到重视，但是它们在病毒感染及其诱导的天然免疫应答过程中的作用和作用机制仍不完全清楚。为此，本项目筛选并鉴定了三条新的功能性lncRNAs，分别命名为lincRNA-up30、lncRNA-155 和lncRNA ISR，并发现它们与流感病毒致病过程密切相关。进一步从分子、细胞和动物水平上对它们的功能和作用机制进行了深入研究，揭示并阐释了这些新型功能性lncRNAs 在宿主抗流感病毒感染与复制中的作用及其内在的分子基础。本研究发现流感病毒通过RIG-I介导的信号通路诱导lincRNA-up30高表达，而高表达的lincRNA-up30通过调控几种关键的干扰素刺激基因（ISGs）的表达水平来影响病毒复制；我们研究揭示了lncRNA-155通过抑制干扰素（IFN）信号通路负调节因子PTP1B的表达，从而上调IFN-β及其下游重要ISGs的水平，促进宿主天然免疫系统发挥抗流感病毒感染的功能；研究探明了lncRNA ISR是由RIG-I依赖性天然免疫信号通路所调节，通过直接调控IFN-β的产生抑制病毒复制，深入研究揭示了在流感病毒感染过程中，lncRNA ISR还通过调节干扰素下游信号通路，在抑制病毒感染和复制过程中发挥重要作用。总之，本项目筛选了三条新的lncRNAs，并完成了对它们的功能验证，通过体外、体内系统实验阐释了这些lncRNAs在宿主抗病毒天然免疫中的作用及其机制，为彻底阐明流感病毒的致病机理提供了重要的科学依据，这些成果将为开发有针对性抗病毒药物提供有价值的新靶标。