Uncovering HIV/opioid effects in the brain at the single cell level: transcription, chromatin accessibility, and reservoir analysis in the SIV/cART/morphine/rhesus monkey model

在单细胞水平上揭示 HIV/阿片类药物对大脑的影响：SIV/cART/吗啡/恒河猴模型中的转录、染色质可及性和储库分析

• 批准号: 10220475
• 负责人: Shilpa J. Buch
• 金额: $ 141.74万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10220475
• 关键词: Address; Affect; American; Animal Model; Animals; Anti-Retroviral Agents; Biological Assay; Biological Markers; Brain; Brain region; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cells; Cellular Assay; Chromatin; Chronic; Controlled Study; Corpus striatum structure; Coupled; Critical Pathways; DNA; Data; Data Set; Disease; Drug abuse; Ensure; Epidemic; Experimental Designs; FAIR principles; Future; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Gold; HIV; HIV Infections; HIV-1; Human; Immune; Immunohistochemistry; In Situ Hybridization; Individual; Infection; Injections; Knowledge; Lymphocyte; Macaca mulatta; Mediating; Microglia; Modeling; Molecular; Morphine; National Institute of Drug Abuse; Neuroglia; Neurons; Neuropathogenesis; Opioid; Pathogenesis; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Phenotype; Population; Predisposition; Prefrontal Cortex; Primates; Protein Isoforms; RNA; Randomized; Reproducibility; Research; Role; SIV; Saline; Sample Size; Site; System; Systems Biology; Techniques; Therapeutic Intervention; Transposase; Twin Multiple Birth; Validation; Variant; Viral; Viral Genome; Viral Pathogenesis; Viral reservoir; Virus; Work; antiretroviral therapy; base; brain cell; cell type; clinically relevant; cocaine exposure; combinatorial; data mining; design; drug of abuse; epigenetic regulation; epigenomics; experimental group; experimental study; follow-up; insight; interdisciplinary approach; macrophage; member; methamphetamine exposure; monocyte; nonhuman primate; novel; opioid abuse; opioid epidemic; opioid exposure; opioid use; outreach; pandemic disease; power analysis; prevent; programs; response; simian human immunodeficiency virus; single cell technology; single-cell RNA sequencing; transcriptome; transcriptome sequencing; virology

Project Summary A comprehensive interdisciplinary approach proposed to support the Single Cell Opioid Responses in the Context of HIV (SCORCH) Program Expansion: CNS Data Generation for Chronic Opioid, Methamphetamine, and/or Cocaine Exposures RFA from the National Institute on Drug Abuse (NIDA). We will perform studies on the interplay of HIV pathogenesis and opioid abuse in the gold-standard SIV/nonhuman primate system, using single-cell RNA sequencing (scRNAseq) to assess individual cellular transcriptomes. This will give us obtain unparalleled insight into the viral and drug-induced effects on diverse cell types in brain in regions critical for HIV infection and opioid abuse. In addition, we will also focus on primary targets of HIV and drivers of neuropathogenesis, microglia and brain macrophages, using scRNAseq as well as single-cell Assay for Transposase-Accessible Chromatin sequencing (scATACseq) to gain insight into the epigenetic regulation of gene expression at the single cell level. These will be combined with state-of-the-art sensitive assays for virus in the brain. We will ensure our data adhere to the FAIR (Findable, Accessible, Interoperable, and Reusable) standards, and will work closely with other members of the SCORCH program to share and disseminate data to yield maximal advancement to the field. Our experiments are carefully designed to address the role of virus, drugs, and their interactions, as well as include the clinically relevant scenarios of those taking antiretrovirals to suppress virus, and those who do not. Our rigor and reproducibility are facilitated by our detailed power analysis with proper sample sizes, as well as our experimental design to match the standards of blocking, replication and randomization. The results from these studies will provide novel insights into the CNS effects of HIV and drug abuse.

项目摘要 一个全面的跨学科的方法，提出了支持单细胞阿片样物质的反应， HIV（SCORCH）计划扩展的背景：慢性阿片类药物、甲基苯丙胺、 和/或来自国家药物滥用研究所（NIDA）的Coconut Exposures RFA。我们将研究 在金标准SIV/非人灵长类系统中，HIV发病机制和阿片类药物滥用的相互作用， 单细胞RNA测序（scRNAseq）以评估单个细胞转录组。这将使我们获得 对病毒和药物诱导的对大脑中对艾滋病毒至关重要的区域的不同细胞类型的影响的无与伦比的洞察力 感染和阿片类药物滥用。此外，我们还将重点关注艾滋病毒的主要目标和 神经发病机制，小胶质细胞和脑巨噬细胞，使用scRNAseq以及用于 转座酶可重复染色质测序（scATACseq），以深入了解表观遗传调控， 在单细胞水平上的基因表达。这些将与最先进的病毒敏感检测相结合 在大脑中。我们将确保我们的数据符合FAIR（可查找、可解释、可互操作和可重用）标准。 标准，并将与SCORCH计划的其他成员密切合作，共享和传播数据， 最大限度地促进了该领域的发展。我们的实验经过精心设计， 药物及其相互作用，以及包括那些服用抗逆转录病毒药物， 抑制病毒和那些不抑制病毒。我们的严谨性和可重复性得益于我们详细的功率分析 适当的样本量，以及我们的实验设计，以匹配标准的阻断，复制和 随机化这些研究的结果将为HIV和药物对CNS的影响提供新的见解。 虐待