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: 8255929
• 负责人: Jian Zhu
• 金额: $ 5.42万
• 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 2014-07-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8255929
• 关键词: Acquired Immunodeficiency Syndrome; Antiviral Agents; Antiviral Response; Bioinformatics; Biology; CD4 Positive T Lymphocytes; Cells; Cellular biology; Disease; Gene Expression; Genes; Goals; HIV; HIV-1; Hela Cells; Human; Immune response; Infection; Integration Host Factors; Interferon Type I; Interferons; Knowledge; Libraries; Maps; Mediating; Molecular Profiling; Ontology; Pathway Analysis; Pathway interactions; Pharmaceutical Preparations; Probability; Proteins; RNA Interference; Research; Resistance; Resources; Role; Screening procedure; Small Interfering RNA; Testing; Vaccines; Validation; Viral; Virus; Virus Diseases; Virus Replication; functional genomics; genome-wide; insight; macrophage; novel; pathogen; prevent; protein protein interaction; 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. PUBLIC HEALTH RELEVANCE: Although there are several effective drugs for treating AIDS/HIV that can slow the course of the disease, there is still no cure or vaccine. We propose to use functional genomic approaches to identify novel HIV restriction factors, and further validate and study their function in physiologically relevant, HIV-infected host cells. The discovery and functional studies of novel HIV restriction factors will help unravel the previously unrecognized cellular antiviral mechanisms that can be used in new strategies to prevent or treat HIV infection.

描述（由申请人提供）：HIV 需要宿主机制来复制。反过来，宿主细胞利用抗病毒因子来保护自身。充分了解 HIV 复制涉及的宿主因素将为我们提供有关 HIV 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 文库进行的功能筛选表明数百种宿主蛋白​​是 HIV 复制的调节剂。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 复制的先天宿主蛋白。该项目的其余目标是专注于进一步验证 以及屏幕中限制因素的功能研究。目标 1 是对通过全基因组 siRNA 筛选确定的 HIV-1 限制因子进行验证和功能分析。我们最近使用多个 RNAi 资源（三个直系同源 si​​RNA 文库）完成了 siRNA 筛选。筛选确定了 212 个潜在的宿主限制因素。为此，我们计划重点关注一系列潜在的抗艾滋病毒宿主因子，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 敲低功效、RNAi 效应拯救、人类原代细胞验证以及一些有前景的 HIV-1 限制因子的机制研究。此外，我们计划测试已确定的限制因子对其他病毒的抑制效果。目标 2 是通过全基因组 siRNA 筛选来鉴定介导干扰素诱导的 HIV-1 限制的因素。人们早就知道，I 型干扰素 (IFN) 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 复制的早期和晚期步骤均可被 IFN-1 阻断。为此，我们将进行全基因组 siRNA 筛选，以确定 IFN 所需活性的 HIV-1 限制因子。通过进行全面的生物信息分析，进一步筛选候选限制因素。将选择一组潜在的 IFN 诱导的 HIV-1 限制因子进行进一步验证和功能研究。我们相信，我们对 HIV-1 限制因素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。 公众健康相关性：尽管有多种治疗艾滋病/艾滋病毒的有效药物可以减缓病程，但仍然没有治愈方法或疫苗。我们建议使用功能基因组方法来识别新的 HIV 限制因子，并进一步验证和研究它们在生理相关的 HIV 感染宿主细胞中的功能。新型艾滋病毒限制因子的发现和功能研究将有助于揭示以前未被认识的细胞抗病毒机制，这些机制可用于预防或治疗艾滋病毒感染的新策略。