Role of proviral loci in HIV latency

前病毒位点在 HIV 潜伏期中的作用

• 批准号: 9330768
• 负责人: JAMES Ivan MULLINS
• 金额: $ 85.22万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-11 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9330768
• 关键词: Address; Aftercare; Blood; CD4 Positive T Lymphocytes; CRISPR/Cas technology; Cell Proliferation; Cell physiology; Cell surface; Cells; Cellular Structures; Clinic; DNA Methylation; Epigenetic Process; Gene Expression; Genes; Genetic; Genetic Transcription; Genome; Gut associated lymphoid tissue; HIV; HIV Infections; Human; Immunologics; Immunosuppressive Agents; Individual; Infection; International; Interruption; Lead; Learning; Maintenance; Methods; Mississippi; Participant; Persons; Phenotype; Phylogenetic Analysis; Primary Infection; Proteome; Provirus Integration; Proviruses; RNA; Regulatory T-Lymphocyte; Role; Site; Source; Specimen; Structure; Suspensions; T-Lymphocyte Subsets; Technology; Time; Tissues; Transcript; Viral; Viral Proteins; Virus; Virus Latency; Virus Replication; cell type; cohort; cytokine; density; experience; in vitro Model; in vivo; insight; integration site; methylation pattern; novel; novel strategies; protein expression; public health relevance; transcriptome; viral rebound; virology

 DESCRIPTION (provided by applicant): HIV persists for many years during therapy and normally rebounds to pretreatment levels when therapy is stopped. However, a small number of individuals have been able to control their infection, despite the presence of infectious virus, fo many years after stopping therapy. Understanding the factors that lead to virus control under these circumstances is critical to our ability to achieve this end in others as well as for an eventual cure of infection. We are learning that where the virus integrates its genome into the infected cell genome is of significance to the maintenance of HIV reservoirs. However, during suppressive therapy, the rarity of cells with replication competent HIV and the lack of expression of viral proteins by these cells have hampered the study of HIV reservoirs. We propose novel methods to overcome the problem of cell rarity by the use of a novel method to enrich for and expand single infected cells for study of structure and RNA expression of the integrated proviral genome and surrounding sequences within the host cell genome. To address the problem of the lack of protein expression, we will use additional novel methods to purify expanded infected cells, as lysates, allowing analysis of the entire cellular transcriptome and proteome. Lastly, we will recreate the proviral structures we observe in vivo in an in vitro model to discern the global impact of provirus integration on cell functionality.

 描述（由申请人提供）：HIV 在治疗期间持续存在多年，并且在治疗停止时通常会反弹至治疗前的水平。然而，尽管存在传染性病毒，少数人在停止治疗多年后仍然能够控制感染。了解在这种情况下导致病毒控制的因素对于我们在其他情况下实现这一目标以及最终治愈感染的能力至关重要。我们了解到，病毒将其基因组整合到受感染细胞基因组中的位置对于艾滋病毒储存库的维持具有重要意义。然而，在抑制治疗期间，具有复制能力的HIV细胞的稀有以及这些细胞缺乏病毒蛋白的表达阻碍了HIV储存库的研究。我们提出了克服细胞稀有问题的新方法，通过使用一种新方法来富集和扩展单个感染细胞，以研究宿主细胞基因组内整合的原病毒基因组和周围序列的结构和RNA表达。为了解决蛋白质表达缺乏的问题，我们将使用其他新方法来纯化扩增的感染细胞（作为裂解物），从而可以分析整个细胞转录组和蛋白质组。最后，我们将在体外模型中重建我们在体内观察到的前病毒结构，以辨别全局 原病毒整合对细胞功能的影响。