HERV proteogenomics of narcotic-driven HIV latency

麻醉药驱动的 HIV 潜伏期的 HERV 蛋白质基因组学

• 批准号: 10675341
• 负责人: LUBBERTUS C MULDER
• 金额: $ 72.25万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675341
• 关键词: Affect; Anatomy; Autopsy; Base Pairing; Biology; Brain; CD4 Positive T Lymphocytes; CRISPR interference; CRISPR-mediated transcriptional activation; Cell Differentiation process; Cell Line; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Complex; Conserved Sequence; DNA Transposable Elements; Data; Data Set; Detection; Elements; Endogenous Retroviruses; Epigenetic Process; Event; Exposure to; Gene Expression Profile; Genetic Transcription; Genome; Genomics; Goals; HIV; HIV Infections; HIV-1; HIV-associated neurocognitive disorder; Human; Human Characteristics; Human Genome; Immune system; Immunology; In Situ Hybridization; Individual; Infection; Investigation; Knowledge; Link; Maps; Mass Spectrum Analysis; Methodology; Microglia; Mission; Molecular; Narcotics; Nature; Opiate Addiction; Opioid; Patients; Peptides; Persons; Phase; Phenotype; Play; Productivity; Proviruses; RNA; Recording of previous events; Reporter; Reporting; Resolution; Retrotransposon; Retrovirus Proteins; Role; Shapes; Site; Sorting; Source; Substance Use Disorder; Substance abuse problem; Technology; Testing; Therapeutic Agents; Time; Tissues; Virus; Virus Diseases; Work; brain tissue; cancer type; cell immortalization; cell type; deep sequencing; differential expression; high reward; high risk; induced pluripotent stem cell; innovation; insight; integration site; multidisciplinary; opioid exposure; protein expression; proteogenomics; transcriptome sequencing; vector; virology

Abstract HIV eradication requires the elimination of latent reservoirs composed by cells harboring persistent proviruses. The brain is an anatomical site that is poorly accessible to the immune system as well as therapeutic agents. It also harbors a specific type of cells, microglia, where HIV can silently persist for long periods of time. Human endogenous retroviruses (HERVs) are a group of transposable elements, which are mostly held in an inactive state by different epigenetic mechanisms. Narcotic use induces changes the epigenetic landscape that can influence latency establishment and HERV expression. This project’s goal is to test the hypothesis that the distinct HERV expression signatures detected in HIV LTR-silent infection affect latency establishment and that this can be influenced by narcotic treatment. Furthermore, latent HIV-specific HERV expression can be exploited for the discovery of HIV latency specific markers. In order to investigate these hypotheses, it is crucial to determine the precise identity of the HERV elements expressed during latency. The experimental strategy is based on our newly developed reporter virus, which distinguishes latently infected from productively infected microglia cells in combination with our established proteogenomic approach which integrates HERV-specific RNA expression analysis with mass spec analysis. Such highly specialized processing is required because of the repetitive nature of HERVs that so far has undermined their detailed investigation. The RNA and protein expression blueprint obtained will inform on the ranking of the latent HIV- specific HERVs whose relevance will be investigated utilizing state of the art CRISPR-based expression induction in primary microglia as well as for the detection their expression in brain tissue form HIV patients with and without documented history on narcotic use. We expect that the innovative experimental approach proposed will provide insight into HERV RNA/protein expression landscapes in latently infected primary microglia and offer potential causal links between latency establishment, HERV expression and substance abuse. Even though there are high-risk aspects in this proposal, the experimental strategy proposed will deliver predictable, high gain benefits in our understanding of the complex biology underlying HIV latency in the brain compartment.

摘要 根除艾滋病毒需要消除由携带持久性前病毒的细胞组成的潜伏宿主。 大脑是一个很难被免疫系统和治疗剂访问的解剖位置。它 还含有一种特定类型的细胞，即小胶质细胞，在那里艾滋病毒可以静静地持续很长一段时间。 人类内源性逆转录病毒(HERV)是一组转座元件，主要存在于 不同的表观遗传机制所致的非活跃态。麻醉剂的使用导致了表观遗传格局的改变 会影响潜伏期的建立和HERV的表达。这个项目的目标是检验这样一个假设 在HIV LTR沉默感染中检测到不同的Herv表达特征会影响潜伏期的建立，并且 这可能会受到麻醉治疗的影响。此外，潜伏的HIV特异性HERV表达可以 被用来发现艾滋病毒潜伏期特异性标记。为了研究这些假说， 对于确定潜伏期期间表达的HERV元件的准确身份至关重要。 实验策略是基于我们新开发的报告病毒，它区分潜伏感染的 与我们已建立的蛋白质组学方法相结合 将HERV特异性RNA表达分析与质谱仪分析相结合。如此高度专业化 处理是必需的，因为HERV的重复性到目前为止已经破坏了其详细的 调查。所获得的RNA和蛋白质表达蓝图将对潜伏的HIV- 将利用基于CRISPR的最新表达来研究其相关性的特定HERV HIV感染者原代小胶质细胞的诱导及其在脑组织中的表达 而且没有记录的麻醉药品使用历史。我们期待创新的实验方法 建议将深入了解HERV RNA/蛋白质在潜伏感染的原发病例中的表达情况 并提供潜伏期建立、HERV表达和物质之间的潜在因果联系 虐待。 尽管这项提议中有高风险的方面，但所提议的实验策略将产生 可预测的高增益有助于我们理解大脑中潜伏的艾滋病毒的复杂生物学 车厢。