Characterizing Vpr-mediated Activation of the DNA Damage Response (DDR) in HIV Replication

HIV 复制中 Vpr 介导的 DNA 损伤反应 (DDR) 激活的特征

• 批准号: 10613920
• 负责人: Carina Sandoval
• 金额: --
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-07-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10613920
• 关键词: Acquired Immunodeficiency Syndrome; BIRC3 gene; Biological Assay; Biology; Cell Cycle Arrest; Cells; Collaborations; Comet Assay; Complex; Consequences of HIV; DNA Damage; DNA Replication Damage; DNA Viruses; Data; Dendritic Cells; Development; Genes; Genetic Transcription; Genome; Goals; HIV; HIV Infections; HIV-1; HIV-2; IL8 gene; Immune; Immunofluorescence Immunologic; Infection; Knock-out; Knowledge; Lentivirus; Letters; Life Cycle Stages; Link; M cell; Macrophage; Mediating; Molecular; Molecular Biology; Mutation Analysis; NBS1 gene; NF-kappa B; Natural Immunity; Nuclear; Pathway interactions; Persons; Phenotype; Primate Lentiviruses; Quantitative Reverse Transcriptase PCR; RNA Viruses; Repression; Research; Role; Scientist; Signal Pathway; Signal Transduction; Testing; Up-Regulation; Viral; Virus; Virus Replication; Virus-like particle; antiretroviral therapy; assault; genotoxicity; global health; improved; in vivo; inhibitor; insight; knock-down; live cell imaging; monocyte; novel; p53-binding protein 1; p65; recruit; response; success; transcriptome sequencing; tumorigenesis; vpr Gene Products

PROJECT SUMMARY HIV continues to be a major burden to global health as it accounts for approximately 36.9 million people living with acquired immunodeficiency syndrome (AIDS). Scientists have made considerable strides in understanding the basic molecular biology of HIV, which directly led to the development of effective antiretroviral therapy (ART). Yet, despite years of research, we currently do not have a complete picture of the HIV lifecycle to help develop a functional cure. By understanding key steps in the HIV lifecycle and establishing the cellular consequences of HIV infection, we can develop novel treatments and a functional cure, which take advantage of the cellular markers that HIV infection leaves behind. One key aspect of HIV biology that remains unclear is Vpr, an accessory gene without a known primary conserved function. Vpr is evolutionarily conserved among extant primate lentiviruses and is required for virus replication in vivo. Although Vpr enhances viral replication in monocyte-derived macrophages and monocyte-derived dendritic cells, it remains unclear how Vpr enhances replication. An emerging phenotype described for Vpr is engagement with the DNA damage response (DDR). The DDR is a signaling cascade responsible for safeguarding the genome from genotoxic assault that is well understood. Modulation of the DDR is often seen by diverse RNA and DNA viruses to promote tumorigenesis or enhancement of viral replication. Most interestingly, we found that Vpr can induce DNA damage and the ability of Vpr to induce DNA damage is conserved among HIV-1 and HIV-2. In addition, activation of the DDR is linked to NF-kB and innate immunity. Our preliminary data suggest that Vpr-induced DNA damage activates NF-kB. This data contributes to our central hypothesis that Vpr-induced DNA damage signals through the classical DDR to activate NF-kB target genes and enhance viral replication. In this proposal, we will investigate how and why Vpr induces DNA damage. In Aim 1, I will characterize the requirements and downstream signaling of Vpr-induced DNA damage. Completing this aim will shed light on how Vpr induces DNA damage and establishes if the classical DDR is activated or if non-classical activation mechanisms await to be explored. In Aim 2, I will identify the role of Vpr-induced DNA damage activation of NF- κB target genes in HIV replication. Completing this aim will determine why Vpr causes DNA damage and will establish the molecular mechanism for Vpr-enhancement of viral replication. Overall, we will gain insight into how HIV infected cells are dealing with DNA damage. The long-term goal is to leverage our understanding of DNA damage in HIV infected cells to develop improved treatments and a functional cure for people with HIV/AIDS.

项目总结 艾滋病毒仍然是全球卫生的主要负担，因为它约有3690万人生活在 患有获得性免疫缺陷综合症(艾滋病)。科学家们在理解 艾滋病毒的基本分子生物学，这直接导致了有效的抗逆转录病毒疗法(ART)的发展。 然而，尽管多年来的研究，我们目前还没有一个完整的艾滋病毒生命周期的图景来帮助开发 一种有效的治疗方法。通过了解艾滋病毒生命周期中的关键步骤并确定 HIV感染，我们可以开发新的治疗方法和功能疗法，这利用了细胞 艾滋病毒感染留下的标志物。HIV生物学中仍不清楚的一个关键方面是VPR，一种 没有已知的初级保守功能的辅助基因。VPR在进化上是在现存的 灵长类慢病毒，是体内病毒复制所必需的。尽管VPR增强了病毒的复制 单核细胞来源的巨噬细胞和单核细胞来源的树突状细胞，目前尚不清楚VPR是如何增强的 复制。VPR的一个新的表型是与DNA损伤反应(DDR)的结合。 DDR是一个信号级联反应，负责保护基因组免受基因毒物的攻击 明白了。DDR的调节通常被不同的RNA和DNA病毒看到，以促进肿瘤的发生或 加强病毒复制。最有趣的是，我们发现VPR可以诱导DNA损伤和 VPR诱导DNA损伤的机制在HIV-1和HIV-2中是保守的。此外，DDR的激活也与 核因子-kB和先天免疫。我们的初步数据表明，VPR诱导的DNA损伤激活了核因子-kB。 这一数据支持了我们的中心假设，即VPR通过经典的 DDR激活核因子-kB靶基因，增强病毒复制。 在这项提案中，我们将调查VPR如何以及为什么会导致DNA损伤。在目标1中，我将描述 VPR诱导DNA损伤的要求和下游信号转导。完成这一目标将有助于 VPR如何诱导DNA损伤以及确定经典DDR是否被激活或非经典激活 其机制有待探索。在目标2中，我将确定VPR诱导的核因子-2的DNA损伤激活的作用。 κB靶基因在艾滋病毒复制中的作用完成这一目标将确定VPR为什么会导致DNA损伤并将 建立VPR促进病毒复制的分子机制。总体而言，我们将深入了解 HIV感染细胞如何处理DNA损伤。长期目标是利用我们对 HIV感染细胞的DNA损伤，以开发改进的治疗方法和功能性治疗方法 艾滋病毒/艾滋病。