Identification of HIV-1 Restriction Factors through Functional Genomic Screens

通过功能基因组筛选鉴定 HIV-1 限制因素

• 批准号: 8432608
• 负责人: Jian Zhu
• 金额: $ 1.59万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-14 至 2013-10-01
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8432608
• 关键词: Acquired Immunodeficiency Syndrome; Antiviral Agents; Antiviral Response; Bioinformatics; Biology; CD4 Positive T Lymphocytes; Cells; Cellular biology; Data; Disease; Eligibility Determination; Ensure; Fellowship; Future; Gene Expression; Genes; Genetic Epistasis; Goals; HIV; HIV-1; Hela Cells; Hepatitis C virus; Human; Immune response; Infection; Influenza; Integration Host Factors; Interferon Type I; Interferons; Knowledge; Libraries; Maps; Mediating; Molecular Profiling; Ontology; Pathway Analysis; Pathway interactions; Pharmaceutical Preparations; Probability; Proteins; RNA Interference; Recording of previous events; Research; Resistance; Resources; Role; Small Interfering RNA; Solid; Testing; Vaccines; Validation; Viral; Virus; Virus Diseases; Virus Replication; West Nile virus; functional genomics; genome-wide; insight; macrophage; novel; overexpression; pathogen; prevent; protein protein interaction; research study; screening; virology; virus host interaction

DESCRIPTION (provided by applicant): HIV requires host machinery to replicate. In turn, the host cell protects itself using antiviral factors. A full understanding of the host factors involve in HIV replication will provide us with valuable knowledge of HIV virology and cell biology. It will also help to develop new anti-HIV therapy. Functional screens using genome-wide siRNA libraries have implicated hundreds of host proteins as modulators of HIV replication. In this proposed project, we will use multiple RNAi resources to identify the innate host proteins that restrict HIV-1 replication. The remaining goals for this project are to focus on further validation and functional study of the restriction factors that come out of screens. Aim 1 is the validation and functional analysis of HIV-1 restriction factors identified through genome-wide siRNA screens. We recently completed siRNA screens using multiple RNAi resources (three orthologous siRNA libraries). The screens identified 212 potential host restriction factors. For this aim, we plan to focus on a shortened list of potential anti-HIV host factors, and perform further validation and functional studies to confirm their roles. The plan includes confirmation of siRNA knockdown efficacy, RNAi effect rescue, validation in human primary cells, and mechanistic studies of a few of promising HIV-1 restriction factors. In addition, we plan to test the inhibitory efforts of confirmed restriction factors on other viruses. Aim 2 is the identificatin of factors that mediate interferon- induced restriction of HIV-1 through genome-wide siRNA screens. It has been long known that Type I interferon (IFN) can induce the expression of a large set of interferon-stimulated genes and establish a potent antiviral response. Both early and late steps of HIV-1 replication can be blocked by IFN-1. In this aim, we will perform genome-wide siRNA screens to identify HIV-1 restriction factors whose activity is required by IFN. The candidate restriction factors will be further filtered by performing comprehensive bioinformatic analysis. A set of potential IFN-induced HIV-1 restriction factors will be selected for further validation and functional studies. We believe that our study of HIV-1 restriction factors will provide new insights into the barriers to viral infection and may have important ramifications for the identification of new antiviral targets.

描述(申请人提供)：艾滋病毒需要宿主机器进行复制。反过来，宿主细胞使用抗病毒因子来保护自己。充分了解艾滋病毒复制所涉及的宿主因素将为我们提供有关艾滋病毒病毒学和细胞生物学的宝贵知识。它还将有助于开发新的抗艾滋病毒疗法。使用全基因组siRNA文库的功能筛选表明，数百种宿主蛋白是艾滋病毒复制的调节器。在这个拟议的项目中，我们将使用多种RNAi资源来识别限制HIV-1复制的固有宿主蛋白。该项目的剩余目标是专注于进一步的验证 以及筛选出来的制约因素的功能研究。目的1是通过全基因组siRNA筛选确定的HIV-1限制因子的验证和功能分析。我们最近使用多个RNAi资源(三个同源siRNA文库)完成了siRNA筛选。筛选确定了212个潜在的寄主限制因素。为此，我们计划专注于一系列潜在的抗HIV宿主因子，并进行进一步的验证和功能研究，以确认它们的作用。该计划包括确认 SiRNA敲除效率，RNAi效应拯救，在人类原代细胞中的验证，以及几个有希望的HIV-1限制因子的机制研究。此外，我们计划测试已证实的限制因子对其他病毒的抑制作用。目的2是通过全基因组siRNA筛选鉴定介导干扰素诱导的HIV-1限制的因素。众所周知，I型干扰素可以诱导大量干扰素刺激基因的表达，并建立有效的抗病毒反应。HIV-1复制的早期和晚期都可以被干扰素-1阻断。为此，我们将进行全基因组siRNA筛选，以确定干扰素所需的HIV-1限制因子的活性。通过进行全面的生物信息学分析，将进一步筛选候选限制因素。一组潜在的干扰素诱导的HIV-1限制因子将被选择用于进一步的验证和功能研究。我们相信，我们对HIV-1限制因子的研究将为病毒感染的障碍提供新的见解，并可能对识别新的抗病毒靶点产生重要影响。