Role of PARP-1 in HIV-1 latent infection

PARP-1 在 HIV-1 潜伏感染中的作用

• 批准号: 9207777
• 负责人: Manuel Llano
• 金额: $ 37.75万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9207777
• 关键词: Active Sites; Affect; Binding; CCL19 gene; CD4 Positive T Lymphocytes; CXCR4 gene; Cells; Collaborations; Complement; DNA Integration; Data; Environment; Enzymes; Fibrinogen; Gene Expression; Genes; Genetic Transcription; Goals; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Half-Life; Human; Impairment; Infection; Knock-out; Laboratories; Life Cycle Stages; Lymphoid; Maintenance; Mediating; Modeling; Molecular; NF-kappa B; Organ; Pathogenesis; Patients; Pharmacology; Phase; Play; Poly(ADP-ribose) Polymerases; Positioning Attribute; Predisposition; Process; Production; Receptor Signaling; Recruitment Activity; Research; Rest; Role; Signal Transduction; T memory cell; T-Cell Activation; T-Lymphocyte; Tertiary Protein Structure; Time; Transcription Factor AP-1; Vesicular stomatitis Indiana virus; Viral; Virus; Virus Replication; Work; Zinc; Zinc Fingers; base; cofactor; cytokine; experience; experimental study; in vivo; inhibitor/antagonist; knock-down; latent infection; memory CD4 T lymphocyte; mutant; new therapeutic target; novel; nuclear factors of activated T-cells; permissiveness; public health relevance; receptor; small molecule; transcription factor

 DESCRIPTION (provided by applicant): Our long-term goal is to contribute to the eradication of HIV infection by finding strategies to eliminate latency. HIV-1 infection escapes eradication by establishing a latent reservoir. Consequently, identification of cellular factors implicated in the establishment or maintenance of latency, which is the focus of this application, could reveal new therapeutic targets for suppression of HIV-1 infection. We have recently discovered a novel role of PARP-1 in silencing HIV-1 gene expression in human CD4+ T cells. Our data indicated that PARP-1 knockdown (KD) CD4+ T cells are up to 90-fold more permissive to HIV-1 replication than control cells. Furthermore, re-expression of PARP-1 in the KD cells substantially diminished their susceptibility to HIV-1. In addition, a small molecule that targets the zinc finge domains of PARP-1, but not inhibitors binding to the active site of this enzyme, also increased viral replication in CD4+ T cells by 60-folds. Importantly, HIV-1 DNA integration or the production phase of the HIV-1 life cycle were not affected by PARP-1 deficiency or pharmacological interference, indicating that PARP-1 affects HIV-1 replication at a post-integration step, more likely at the level of gene expression. Notably, the effect of PARP-1 required CD4/CXCR4-mediated viral entry; i.e. PARP-1 did not affect infection by VSV-G pseudotyped viruses. Based on these results, we envision that PARP-1 negatively regulates HIV Env-induced CD4/CXCR4 signaling, limiting in this manner the availability of specific transcription factors and consequently favoring the establishment of latency. Multiple findings support our central hypothesis: (1) HIV Env-induced CD4/co-receptor signaling increases expression of cellular factors (i.e. NF-ATs, AP-1, and NF-kB) that promote viral replication. (2) PARP-1 negatively regulates the transcription of multiple genes induced by T cell activation, in part through inhibition of NF-AT- or NF-kB-dependent transcription. (3) HIV-1 gene expression is largely influenced by the availability of specific cellular transcription factors and latency is favored in transcription factor-deprived cells. We are well positioned to carry out the proposed studies because of our extensive experience in the characterization of the molecular mechanisms of cellular cofactors in HIV replication, and in particular in PARP-1. Collaborations established with experts in HIV-1 latency and T cell signaling fields will complement and very efficiently synergize with our own expertise in these fields. At the conclusion of this research we expect to have defined the role of PARP-1 in HIV latency and better identified its mechanism of action. The work proposed is important because new therapeutic targets to eradicate HIV infection could be discovered.

 描述（由申请人提供）：我们的长期目标是通过寻找消除潜伏期的策略来促进根除HIV感染。HIV-1感染逃脱根除， 形成潜在的储层。因此，鉴定与细胞因子有关的细胞因子， 潜伏期的建立或维持（这是本申请的焦点）可以揭示抑制HIV-1感染的新的治疗靶点。我们最近发现了PARP-1在人类CD 4 + T细胞中沉默HIV-1基因表达的新作用。我们的数据表明，PARP-1敲低（KD）CD 4 + T细胞对HIV-1复制的容许性是对照细胞的90倍。此外，KD细胞中PARP-1的再表达显著降低了它们对HIV-1的易感性。此外，靶向PARP-1的锌指结构域的小分子，但不是与这种酶的活性位点结合的抑制剂，也使CD 4 + T细胞中的病毒复制增加了60倍。重要的是，HIV-1 DNA整合或HIV-1生命周期的生产阶段不受PARP-1缺陷或药理学干扰的影响，表明PARP-1在整合后步骤影响HIV-1复制，更可能是在基因表达水平。值得注意的是，PARP-1的作用需要CD 4/CXCR 4介导的病毒进入;即PARP-1不影响VSV-G假型病毒的感染。基于这些结果，我们设想PARP-1负调节HIV Env诱导的CD 4/CXCR 4信号传导，以这种方式限制特异性转录因子的可用性，从而有利于潜伏期的建立。多项发现支持我们的中心假设：（1）HIV Env诱导的CD 4/共受体信号传导增加了促进病毒复制的细胞因子（即NF-AT、AP-1和NF-kB）的表达。(2)PARP-1部分通过抑制NF-AT-或NF-kB-依赖性转录，负调节T细胞活化诱导的多个基因的转录。(3)HIV-1基因表达在很大程度上受特定细胞转录因子的可用性的影响，并且潜伏期是 在缺乏转录因子的细胞中更受欢迎。由于我们在HIV复制中细胞辅因子的分子机制表征方面具有丰富的经验，特别是在PARP-1中，我们能够很好地进行拟议的研究。与HIV-1潜伏期和T细胞信号传导领域的专家建立的合作将与我们在这些领域的专业知识互补并非常有效地协同。在这项研究的结论，我们 我们希望能够确定PARP-1在HIV潜伏期中的作用，并更好地确定其作用机制。这项工作很重要，因为可以发现根除艾滋病毒感染的新的治疗靶点。