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: 9302262
• 负责人: Haoquan Wu
• 金额: $ 22.95万
• 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/9302262
• 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; HIV therapy; Highly Active Antiretroviral Therapy; Histone Deacetylase Inhibitor; In Vitro; Individual; Integration Host Factors; Investigation; Knock-out; Knowledge; Lead; Libraries; Life Cycle Stages; Light; Maintenance; 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; reactivation from latency; 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）为患者提供长期抑制HIV复制的治疗，但由于长期潜伏的HIV储库-治愈HIV感染的主要障碍之一，因此无法治愈患者。潜伏HIV的再激活一直在深入研究中;然而，由于对潜伏期如何维持的知识有限，尚未开发出专门再激活潜伏HIV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子，并全面了解其机制。在培养细胞中进行全基因组功能缺失筛选的方法一直没有出现，直到最近出现了四篇描述CRISPR-Cas9介导的全基因组敲除筛选的论文。我们独立开发了一种类似的方法来进行全基因组筛选，以确定WNV诱导细胞死亡所不可或缺的宿主因子，结果表明我们的方法可以识别具有高特异性的靶基因。因此，我们有信心，我们可以系统地确定宿主因素，以获得全面的了解宿主因素如何促进HIV潜伏期维持的机制。我们将在不同的J-Lat克隆中测试所鉴定的基因，以鉴定可能特异性和普遍性地重新激活潜伏的HIV的宿主因子，这可能导致根除潜伏的HIV储库的新疗法。