ISG CONTROL OF FLAVIVIRUS INFECTION

ISG 控制黄病毒感染

基本信息

批准号：
8601427
负责人：
SUMIT K CHANDA
金额：
$ 61.32万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8601427
关键词：
Accounting Affect Antiviral Agents Arboviruses Area Attenuated Biochemical Bioinformatics Categories Cell Culture Techniques Cell Line Cells Custom Data Dengue Virus Diamond Disease Disease Outbreaks Ectopic Expression Encephalitis Evaluation Family Flavivirus Flavivirus Infections Genes Genetic Screening Growth Hela Cells Histocompatibility Testing Human Immune Immunity Individual Infection Institutes Interferon Type I Interferons Japanese encephalitis virus Kinetics Knockout Mice Laboratories Libraries Madagascar Modeling Mus Mutation National Institute of Allergy and Infectious Disease Natural Immunity Neurons Outcome Pathogenesis Physiological Population Process Property RNA Viruses Research Personnel Risk Specificity Subfamily lentivirinae Survival Analysis Testing Tissues Tropism Viral Viral Load result Virulence Virulent Virus Virus Diseases Virus Replication West Nile virus base cell type design falls follow-up gene function high throughput screening in vivo insight knock-down loss of function member mouse model novel novel strategies pathogen public health relevance research study response small hairpin RNA stable cell line tissue tropism validation studies

项目摘要

DESCRIPTION (provided by applicant): Members of Flavivirus genus are the most important arthropod-borne viruses causing disease in humans. This genus includes viruses (West Nile virus (WNV), Japanese encephalitis virus (JEV) and Dengue virus (DENV)) that are re-emerging and becoming endemic in new areas of the world. Flaviviruses account for ~100 millions infections per year, with billions at risk and no specific therapy available. Although interferon (IFN) responses control the cell and tissue tropism of WNV and other flaviviruses, the specific effector molecules that restrict infection remain poorly characterized. The studies in this collaborative and inter-disciplinary project between the Diamond and Chanda laboratories will use genetic screens to identify novel interferon stimulated genes (ISG) that modulate flavivirus infection in specific cell types ex vivo and in vivo. Loss-of-function high-throughput genetic screens will be performed with a custom- generated GFP-marked shRNA library targeting ~700 mouse and human ISGs, to identify novel ISG that restrict infection of virulent and attenuated strains of WNV. Using stable cell lines that have targeted reductions or ectopic expression of candidate ISGs, we will define mechanistically how novel ISG effector molecules restrict specific steps in the viral lifecycle and influence infection outcome. In preliminary studies, we have identified several candidate inhibitory ISG (Ifi27, IFIT2, and IFITM3) that attenuate WNV infection, and already acquired or generated knockout mice. Using these mice, we will evaluate the function of these particular ISGs in restricting WNV replication in a cell-specific manner and in vivo. We hypothesize that specific ISGs have antiviral properties that differentially control WNV infection and spread, and that these demonstrate tissue and cell-type specificity. Overall, these experiments will more clearly define the interface between IFN control and flavivirus pathogenesis and possibly, guide strategies that modulate immunity to infection by this family of viruses.

描述（由申请人提供）：黄病毒属的成员是最重要的节肢动物传播病毒，导致人类疾病。该属包括病毒（西尼罗河病毒（WNV），日本脑炎病毒（JEV）和登革热病毒（DENV）），这些病毒正在重新出现并在世界新领域成为地方性。黄病毒每年占约1亿感染，数十亿人处于危险之中，没有特定的治疗。尽管干扰素（IFN）反应控制了WNV和其他黄素病毒的细胞和组织向量，但限制感染的特定效应分子的特征仍然很差。钻石和木达实验室之间的合作和跨学科项目中的研究将使用遗传筛选来识别新型干扰素刺激基因（ISG），该基因（ISG）调节特定细胞类型的Ex ex ex and Vivo中的黄病毒感染。功能丧失的高通量遗传筛选将使用定制的GFP标记的shRNA库，靶向〜700小鼠和人类ISG，以识别限制WNV毒性和减弱菌株的新型ISG。使用针对候选ISG的稳定细胞系，我们将在机械上定义新型ISG效应分子如何限制病毒生命周期中的特定步骤并影响感染结果。在初步研究中，我们已经确定了减弱WNV感染的几个候选抑制性ISG（IFI27，IFIT2和IFITM3），并且已经获得或产生了敲除小鼠。使用这些小鼠，我们将评估这些特定ISG在以细胞特异性和体内限制WNV复制中的功能。我们假设特定的ISG具有抗病毒特性，可以差异地控制WNV感染和扩散，并且这些特性表现出组织和细胞类型的特异性。总体而言，这些实验将更清楚地定义IFN控制与黄病毒发病机理之间的界面，并可能引导策略调节该病毒家族对感染的免疫感。