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.

项目概要 艾滋病毒仍然是全球健康的主要负担，约 3,690 万人感染艾滋病毒 科学家们对获得性免疫缺陷综合症（艾滋病）的了解已经取得了长足的进步。 HIV 的基本分子生物学，直接导致了有效的抗逆转录病毒疗法（ART）的发展。 然而，尽管经过多年的研究，我们目前还没有全面了解艾滋病毒生命周期，以帮助开发 通过了解艾滋病毒生命周期的关键步骤并确定其细胞后果。 HIV 感染，我们可以开发新的治疗方法和功能性治愈方法，利用细胞 HIV 感染留下的一个仍不清楚的重要特征是 Vpr，它是一种病毒标记物。 没有已知主要保守功能的辅助基因在现存的进化中是保守的。 灵长类慢病毒是病毒体内复制所必需的，尽管 Vpr 可以增强病毒在体内的复制。 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 如何增强 Vpr 的一个新兴表型是与 DNA 损伤反应 (DDR) 相关。 DDR 是一个信号级联，负责保护基因组免受基因毒性攻击，这很好 人们普遍认为，多种 RNA 和 DNA 病毒对 DDR 的调节可促进肿瘤发生或促进肿瘤发生。 最有趣的是，我们发现Vpr具有诱导DNA损伤的能力。 Vpr 诱导 DNA 损伤的机制在 HIV-1 和 HIV-2 中是保守的。此外，DDR 的激活也是相关的。 我们的初步数据表明，Vpr 诱导的 DNA 损伤会激活 NF-kB。 这些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。 在本提案中，我们将研究 Vpr 如何以及为何引起 DNA 损伤。在目标 1 中，我将描述 Vpr 的特征。 Vpr 诱导的 DNA 损伤的要求和下游信号传导将有助于阐明这一目标。 Vpr 如何诱导 DNA 损伤并确定经典 DDR 是否被激活或非经典激活 在目标 2 中，我将确定 Vpr 诱导的 NF-DNA 损伤激活的作用。 HIV 复制中的 κB 靶基因将确定 Vpr 为何会造成 DNA 损伤。 总体而言，我们将深入了解 Vpr 增强病毒复制的分子机制。 长期目标是利用我们对 HIV 感染细胞如何处理 DNA 损伤的理解。 HIV 感染细胞中的 DNA 损伤，旨在为艾滋病患者开发改进的治疗方法和功能性治愈方法 艾滋病毒/艾滋病。