Establishing HIV-1 chromatin in resting T cells: Vpr, latency, and H2A.Z

在静息 T 细胞中建立 HIV-1 染色质：Vpr、潜伏期和 H2A.Z

• 批准号: 10558472
• 负责人: DAVID N LEVY
• 金额: $ 39.63万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10558472
• 关键词: Apoptosis; Binding; Biological Assay; CD4 Positive T Lymphocytes; Cell Cycle Arrest; Cells; Chromatin; Chromatin Remodeling Factor; Chromatin Structure; Chronic; Complex; DNA; DNA Damage; Data; Disputes; Epigenetic Process; Event; Evolution; Gene Expression; Genes; Genetic Transcription; Glean; HIV; HIV Infections; HIV-1; Histone Deacetylation; Histone H2A; Histones; Immediate-Early Proteins; Infection; Lentivirus; Mediating; Modification; Molecular; Nucleosomes; Pathway interactions; Pharmaceutical Preparations; Positive Transcriptional Elongation Factor B; Post-Translational Protein Processing; Process; Promoter Regions; Protein Isoforms; Proteins; Proviruses; RNA; Regulation; Reporter; Repression; Rest; Reverse Transcription; Role; Signal Transduction; Stimulus; Structure; T-Cell Activation; T-Lymphocyte; Testing; Time; Transactivation; Transcriptional Regulation; Transformed Cell Line; Viral; Viral Genome; Viral Proteins; Virion; Virus; Virus Replication; Work; design; epigenetic regulation; histone methylation; insight; latent infection; mutant; novel; permissiveness; preservation; programs; promoter; purge; recruit; response; small molecule inhibitor; therapeutic development; transcriptome sequencing; vpr Gene Products

Project Summary Latent HIV infection of resting CD4 T cells present a formidable challenge in the pursuit of treatments to purge HIV from the body. Latency is established and regulated in large part through the modifications of the nucleosomes that are bound around the viral promoter region. Drugs that reactivate latent HIV must influence these structures in order to allow HIV transcription and virus expression. However, for unknown reasons not all intact proviruses respond to latency reversing agents and thus present a particularly troublesome barrier to cure. A greater understanding of the regulation of HIV-1 chromatin, the installation of nucleosomes and their histone post translational modifications (PTM) will contribute greatly to the development of therapeutic control over HIV- 1 latency. This project seeks such an understanding and builds upon extensive preliminary studies revealing important new insights into these processes. For the first time we investigate the initial stages of HIV-1 chromatin installation, finding that it occurs either contemporaneously or soon after reverse transcription, and before integration into the cellular DNA. We find that repressive chromatin is installed in the absence of the viral protein Vpr being delivered in the virion. Virion Vpr dramatically increases the number of transcriptionally active proviruses in the first 4 days after infection of resting CD4 T cells. When Vpr is present, the nucleosomes that control HIV expression are modified in ways that facilitate HIV replication and block installation of repressive chromatin. For example, the alternate histone H2A.Z is installed only when virion Vpr is available, and H2A.Z is a central organizer of paused but responsive promoters. Without Vpr, the chromatin of latent proviruses takes on a more repressed structure, resulting in more proviruses that do not respond to latency reversing agents. Novel RNA-seq data demonstrate that virion Vpr reprograms gene expression pathways central to chromatin organization and transcriptional regulation. This project will describe both the initial steps in HIV chromatization and the long term processes that lead to reversible latency and irreversible repression. Aim 1 will systematically analyze early proviral chromatin in resting CD4 T cells and the influence of Vpr and Tat-directed transcription on the epigenetic landscape. Our central hypothesis is that Vpr directs installation of the transcriptional pre-initiation complex for basal pre-Tat transcription in resting T cells. Aim 2 will analyze Vpr-dependent pathways leading to these structures. RNA-seq data will be expanded and used to study novel Vpr targets of regulation, and known Vpr pathways will be investigated for their influence on early events. Aim 3 will examine long term infection and latency under the influence the structures and pathways gleaned from Aims 1 and 2. Our hypothesis is that Vpr protects the provirus from epigenetic repression and irreversible latency. The proposed studies will provide much needed information that will assist in the development of therapeutics that can purge HIV from the body or permanently repress virus replication without continual antiviral treatments.

项目摘要 静息CD4 T细胞的潜在艾滋病毒感染在追求治疗以清除时提出了巨大的挑战 来自身体的艾滋病毒。通过修改，在很大程度上建立了潜伏期和监管 与病毒启动子区域结合的核小体。重新激活潜在艾滋病毒的药物必须影响 这些结构以允许HIV转录和病毒表达。但是，出于未知原因并非全部 完整的原病毒应对延迟逆转代理的反应，从而呈现出一个特别麻烦的治愈障碍。 对HIV-1染色质的调节，核小体的安装及其组蛋白的调节有了更大的了解 翻译后修改（PTM）将对HIV-的治疗控制产生产生巨大贡献 1延迟。该项目寻求这种理解，并以广泛的初步研究为基础 对这些过程的重要新见解。我们第一次研究HIV-1染色质的初始阶段 安装，发现它是同时或在逆转录后不久和之前发生的 整合到细胞DNA中。我们发现在没有病毒蛋白的情况下安装抑制性染色质 VPR在病毒座中交付。 Virion VPR急剧增加了转录活性的数量 静止CD4 T细胞感染后的前4天，病毒病毒。当存在VPR时， 控制艾滋病毒表达以促进艾滋病毒复制和阻碍安装的方式进行修改 染色质。例如，仅在可用VIRION VPR时安装备用组蛋白H2A.Z，而H2A.Z为 暂停但响应迅速的启动子的中央组织者。如果没有VPR，潜在病毒的染色质就有 在更受压抑的结构上，导致更多的预科病毒对潜伏期逆转代理的反应。 新型RNA-seq数据表明，Virion VPR重编程基因表达途径与染色质中心 组织和转录监管。该项目将描述HIV色谱的两个初始步骤 以及导致可逆潜伏期和不可逆抑制的长期过程。 AIM 1将系统地 分析早期病毒染色质在静息CD4 T细胞中的早期染色质以及VPR和TAT指导转录的影响 表观遗传景观。我们的中心假设是VPR指导转录预设的安装 复合物用于静息T细胞中的基础前TAT转录。 AIM 2将分析导致VPR依赖性途径 这些结构。 RNA-seq数据将扩展并用于研究新型调节的VPR目标，并已知 VPR途径将因其对早期事件的影响而进行调查。 AIM 3将检查长期感染和 在影响1和2的结构和途径的影响下的潜伏期。我们的假设是VPR 保护病毒免受表观遗传抑制和不可逆转的潜伏期。拟议的研究将提供很多 需要的信息，这些信息将有助于开发可以从身体清除艾滋病毒或 永久抑制病毒复制，而无需连续的抗病毒治疗。

项目成果

Multiple infection of cells changes the dynamics of basic viral evolutionary processes

细胞的多重感染改变了基本病毒进化过程的动态

• DOI: 10.1002/evl3.95
• 发表时间: 2019
• 期刊: Evolution Letters
• 影响因子: 5
• 作者: Wodarz, Dominik;Levy, David N.;Komarova, Natalia L.
• 通讯作者: Komarova, Natalia L.

NNRTI-induced HIV-1 protease-mediated cytotoxicity induces rapid death of CD4 T cells during productive infection and latency reversal

• DOI: 10.1186/s12977-019-0479-9
• 发表时间: 2019-06
• 期刊: Retrovirology
• 影响因子: 3.3
• 作者: B. Trinité;Hongtao Zhang;D. N. Levy

PSGL-1 restricts HIV-1 infectivity by blocking virus particle attachment to target cells

• DOI: 10.1073/pnas.1916054117
• 发表时间: 2020-04-28
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Fu, Yajing;He, Sijia;Wu, Yuntao
• 通讯作者: Wu, Yuntao