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.

抽象的 消除艾滋病毒需要消除由持有持续性病毒的细胞组成的潜在储层。 大脑是一个解剖部位，免疫系统以及治疗剂无法接近。 还拥有一种特定类型的细胞小胶质细胞，其中HIV可以长时间静止地持续存在。 人内源性逆转录病毒（HERV）是一组可替代元素，这些元素主要保存在一个 通过不同的表观遗传机制进行非活性状态。麻醉性使用会导致表观遗传景观改变 可以影响潜伏期的建立和HERV表达。该项目的目标是检验以下假设 在艾滋病毒LTR-SILENT感染中检测到的独特的HERV表达签名会影响潜伏期的建立，并 这可能受麻醉治疗的影响。此外，潜在的HIV特异性HERV表达可能是 被利用用于发现艾滋病毒潜伏期特定标记。为了调查这些假设，是 对于确定潜伏期表达的HERV元素的精确身份至关重要。 实验策略是基于我们新开发的记者病毒，该病毒区分开了。 从有效感染的小胶质细胞与我们既定的蛋白质方法结合 综合HERV特异性RNA表达分析和质量分析。如此专业 需要处理，因为HERV的重复性破坏了他们的细节 调查。获得的RNA和蛋白表达蓝图将告知潜在HIV-的排名 特定的HERV将使用基于CRISPR的最先进的表达来调查其相关性 原代小胶质细胞的诱导以及检测其在脑组织中的表达 并且没有关于麻醉性使用的历史记录。我们期望创新的实验方法 提出的将洞悉潜在受感染的原发性的HERV RNA/蛋白质表达景观 小胶质细胞并提供潜在的因果关系，在潜伏期建立，HERV表达和底物之间 虐待。 即使该提案中有高风险的方面，提出的实验策略也将提供 可预测的，高收益的好处在我们对大脑中艾滋病毒潜伏期的复杂生物学的理解中 车厢。