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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的根除艾滋病毒需要消除由携带持久性原病毒的细胞组成的潜在储存库。大脑是免疫系统和治疗药物很难接触到的解剖部位。它还含有一种特殊类型的细胞，即小胶质细胞，艾滋病毒可以在其中默默地存在很长一段时间。人内源性逆转录病毒（HERV）是一组转座因子，它们大多保存在不同的表观遗传机制导致的非活跃状态会导致表观遗传景观的改变。可以影响潜伏期建立和 HERV 表达该项目的目标是检验以下假设：在 HIV LTR 沉默感染中检测到的不同 HERV 表达特征会影响潜伏期的建立，并且此外，潜伏的 HIV 特异性 HERV 表达可能会受到麻醉治疗的影响。用于发现 HIV 潜伏期特异性标记物为了研究这些假设，我们正在研究这些假设。对于确定潜伏期间表达的 HERV 元件的精确身份至关重要。实验策略基于我们新开发的报告病毒，可以区分潜伏感染的病毒来自有效感染的小胶质细胞与我们建立的蛋白质组学方法相结合，将 HERV 特异性 RNA 表达分析与此类高度专业化的质谱分析相结合。由于 HERV 的重复性，迄今为止已经破坏了其详细信息，因此需要进行处理。获得的 RNA 和蛋白质表达蓝图将告知潜在 HIV 的排名。将利用最先进的基于 CRISPR 的表达来研究特定 HERV 的相关性诱导原代小胶质细胞以及检测其在 HIV 患者脑组织中的表达并且没有关于麻醉剂使用的历史记录，我们期望创新的实验方法。提议将深入了解潜伏感染原发性中的 HERV RNA/蛋白表达情况小胶质细胞并提供潜伏期建立、HERV 表达和物质之间潜在的因果关系虐待。尽管该提案存在高风险，但所提出的实验策略将实现可预测的、高收益的益处有助于我们了解大脑中艾滋病毒潜伏期的复杂生物学隔间。