Identifying host factors essential for HIV latency maintenance

确定 HIV 潜伏期维持所必需的宿主因素

• 批准号: 9136545
• 负责人: Haoquan Wu
• 金额: $ 19.13万
• 依托单位: TEXAS TECH UNIVERSITY HEALTH SCIS CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9136545
• 关键词: Appearance; Biological Process; CD4 Positive T Lymphocytes; Cell Death; Cell model; Cells; Clinical Trials; Cultured Cells; Data Quality; Development; Drug Targeting; Ensure; Gene Silencing; Gene Targeting; Genes; Genome; Genome engineering; Goals; Grant; HIV; HIV Genome; HIV Infections; Highly Active Antiretroviral Therapy; Histone Deacetylase Inhibitor; In Vitro; Individual; Integration Host Factors; Investigation; Knock-out; Knowledge; Lead; Libraries; Life Cycle Stages; Light; Maintenance; Maps; Mediating; Methods; Modeling; Molecular; Paper; Patients; Performance; Phenotype; Proviruses; Public Health; RNA Interference; Reporting; Research; Site; Specificity; System; Technology; Testing; Time; West Nile virus; base; design; endonuclease; genome-wide; genome-wide analysis; improved; insight; latent infection; loss of function; novel strategies; novel therapeutics; public health relevance; screening; small hairpin RNA; therapeutic target; tool

 DESCRIPTION (provided by applicant): Highly active antiretroviral therapy (HAART) offers patients long-term suppression of HIV replication, but it does not cure the patients because of the long lasting latent HIV reservoir- one of the major obstacles in curing HIV infection. Reactivation of latent HIV has been under intensive investigation; however, no strategy has been developed to specifically reactivate latent HIV due to limited knowledge of how the latency is maintained. We propose to perform genome- wide knockout screening to identify the host factors that are essential for HIV latency maintenance and gain comprehensive insight into the mechanism. There has been no method for performing genome-wide loss-of-function screening in cultured cells until the appearance of four recent papers that described a CRISPR-Cas9-mediated genome-wide knockout screen. We have independently developed a similar approach to perform genome-wide screening to identify the host factors that are indispensable for WNV-induced cell death, and the results show that our method can identify target genes with high specificity. Thus, we are confident that we can identify the host factors systematically to gain comprehensive insight into the mechanism of how host factors facilitate HIV latency maintenance. We will test the identified genes in different J-Lat clones to identify the host factors that might reactivate latent HIV specifically and universally, which might lead to a new therapy to eradicate latent HIV reservoir.

 描述（由申请人提供）：高效抗逆转录病毒疗法（HAART）可以为患者提供长期抑制HIV复制的功能，但它无法治愈患者，因为长期潜伏的HIV病毒库是治愈HIV感染的主要障碍之一。潜伏艾滋病毒的重新激活一直在深入研究中；然而，由于对如何维持潜伏期的了解有限，尚未制定专门重新激活潜伏期艾滋病毒的策略。我们建议进行全基因组敲除筛选，以确定对 HIV 潜伏期维持至关重要的宿主因素，并全面了解该机制。直到最近出现四篇描述 CRISPR-Cas9 介导的全基因组敲除筛选的论文之前，还没有在培养细胞中进行全基因组功能丧失筛选的方法。我们独立开发了类似的方法进行全基因组筛选，以确定西尼罗河病毒诱导细胞死亡所必需的宿主因子，结果表明我们的方法可以高度特异性地识别目标基因。因此，我们有信心能够系统地识别宿主因素，以全面了解宿主因素如何促进艾滋病毒潜伏期维持的机制。我们将测试不同 J-Lat 克隆中已识别的基因，以确定可能特异性且普遍地重新激活潜伏 HIV 的宿主因子，这可能会导致一种根除潜伏 HIV 病毒库的新疗法。