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: 10483976
• 负责人: Carina Sandoval
• 金额: $ 4.14万
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10483976
• 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; Light; Link; M cell; 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; 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; macrophage; 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.

项目摘要 艾滋病毒仍然是全球健康的重大烧伤，因为它约占居住的人数约3690万 与获得的免疫缺陷综合征（AIDS）。科学家在理解方面取得了长足的进步 HIV的基本分子生物学直接导致有效的抗逆转录病毒疗法（ART）的发展。 然而，目的地研究多年，我们目前没有艾滋病毒生命周期的完整图片来帮助发展 功能治愈。通过了解HIV生命周期中的关键步骤并确定细胞的后果 艾滋病毒感染，我们可以开发新的治疗方法和功能治愈，从而利用细胞的优势 艾滋病毒感染留下的标记。 HIV生物学的一个关键方面尚不清楚，VPR是 附件基因没有已知的主要配置功能。 VPR在广泛的 灵长类动病毒是体内病毒复制所必需的。尽管VPR增强了病毒复制 单核细胞衍生的巨噬细胞和单核细胞衍生的树突状细胞，尚不清楚VPR如何增强 复制。 VPR描述的新兴表型是与DNA损伤响应（DDR）的参与。 DDR是负责保护基因组免受基因毒性攻击的信号传导级联 理解。 DDR的调节通常是由Driver RNA和DNA病毒观察到的，以促进肿瘤发生或 增强病毒复制。最有趣的是，我们发现VPR可以诱导DNA损伤和能力 在HIV-1和HIV-2中，VPR诱导DNA损伤是保守的。此外，DDR的激活链接 致NF-KB和先天免疫。我们的初步数据表明，VPR诱导的DNA损伤激活NF-KB。 该数据有助于我们的中心假设，即VPR诱导的DNA损伤信号通过经典 DDR激活NF-KB靶基因并增强病毒复制。 在此提案中，我们将研究如何以及为什么VPR诱导DNA损伤。在AIM 1中，我将表征 VPR诱导的DNA损伤的要求和下游信号传导。完成此目标将阐明 VPR如何诱导DNA损伤并确定是否激活了经典的DDR或非古典激活 等待探索的机制。在AIM 2中，我将确定VPR诱导的NF-的DNA损伤激活的作用 HIV复制中的κB靶基因。完成此目标将决定为什么VPR会造成DNA损伤，并且将 建立用于病毒复制的VPR增强的分子机制。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 艾滋病毒感染细胞中的DNA损伤以开发改进的治疗方法，并为患有 艾滋病毒/艾滋病。