Integrative Single Cell isoform and chromatin accessibility Mapping of Chronic Opioid Exposure in Cognitive Brain Areas in HIV

HIV认知脑区慢性阿片类药物暴露的综合单细胞亚型和染色质可及性图谱

• 批准号: 10657960
• 负责人: Teresa A Milner
• 金额: $ 16.76万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-30 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10657960
• 关键词: Absence of pain sensation; Administrative Supplement; Affect; Agonist; Animals; Area; Astrocytes; Award; Biological Process; Brain; Brain region; Cells; Characteristics; Chromatin; Chronic; Cognitive; Computer Analysis; Computing Methodologies; Data; Data Set; Detection; Drug Exposure; Event; Exons; Exposure to; Functional disorder; Future; Gene Expression; Generations; Genes; Genetic Transcription; Genomics; Goals; HIV; Heterogeneity; Hippocampus (Brain); Human; In Situ Hybridization; Individual; Infection; Knowledge; Literature; Maps; Microglia; Modality; Modeling; Monitor; Morphine; Neuroglia; Neurons; Nomenclature; Opiate Addiction; Opioid; Opioid Peptide; Oxycodone; Parents; Peptide Receptor; Persons; Poly A; Poly(A)+ RNA; Population; Process; Protein Isoforms; Publishing; Regulation; Reporting; Research Activity; Research Design; Resolution; Risk; Rodent; SIV; Signaling Molecule; Site; Slide; Stress; Technology; Time; Viral; Work; antiretroviral therapy; base; brain cell; brain health; cell type; classical conditioning; epigenome; epigenomics; experimental study; genome-wide; innovation; interest; molecular rearrangement; mu opioid receptors; nonhuman primate; novel; opioid abuse; opioid exposure; opioid use disorder; parent grant; programs; response; sequencing platform; sex; single cell sequencing; single-cell RNA sequencing; tool; transcriptome; transcriptome sequencing; transcriptomics; transmission process

Opioid driven exacerbations of neuropathological events and alterations in HIV transcription contributing to HIV associated CNS dysfunction are well-reported. Despite years of continuous suppressive antiretroviral therapy (ART), latent HIV persists and finds sanctuary in many of the same brain regions involved in opioid use disorder (OUD) suggesting interactions between HIV and opioids in brain cells. However, there is a sizeable gap in our knowledge on how OUD impacts cellular responses and viral persistence in HIV-infected brain on ART in humans or relevant model organsims. This administrative supplement proposal seeks to generate new topographical data sets from the parent award and evidence at single cell resolution across the hippocampus, a brain region known for predilection for HIV persistence and OUD in non-human primate (NHP). These data will provide an unprecedented cellular landscape of multiple modalities that can be harnessed to develop strategies to limit viral persistence and restore and retain optimal brain health in people living with HIV. In our published and preliminary work we have developed innovative single-cell approaches including Single-cell isoform RNA sequencing (ScISOr-Seq), which enables single-cell long-read RNA sequencing of polyadenylated RNAs across thousands of single cells. In concert these novel sequencing and computational methods, along with ScATAC-Seq for chromatin accessibility, will permit the mapping of cellular gene expression, open chromatin regions, isoforms and the detection of SIV across single-cells of hippocampus. Recent literature supports the presence of HIV in the brain and more specifically in microglia and astrocytes present within the hippocampus. Importantly, this brain region is also involved in associative learning processes for OUD. Moreover, our prior studies in rodent hippocampus have laid the groundwork for the proposed studies by establishing the regional and cell-specific distributions of opioid peptides and receptors as well as related signaling molecules, and how these distributions are impacted by sex, stress and opioid-associated learning. Using new cutting-edge single-cell mulitomic integrated genomics, we propose in this supplemental application to generate multi-dimensional single-cell data sets and define the relevance of such observations in a controlled SIV infected NHP model with and without a different opioid of documented chronic morphine exposure for comparison with the parent study evaluating oxycodone. We also propose to validate any changes observed in single-cell expression in select genes using in situ hybridization. We propose to achieve these goals under the following specific aims: Aim 1. Generation of genome-wide single-cell transcriptomic profiles in the hippocampus in a controlled setting of SIV infection of NHPs monitored on ART and exposed to chronic morphine. Aim 2. Establish genome-wide single-cell epigenomic profiles of the hippocampus in the groups examined in aim 1.

阿片类药物驱动的神经病理事件的加剧和艾滋病毒转录的改变，导致艾滋病毒 相关的中枢神经系统功能障碍已得到很好的报告。尽管多年连续抑制抗逆转录病毒疗法 （艺术），潜在的艾滋病毒持续存在并在许多相同的大脑区域中发现了圣所 （OUD）提示脑细胞中HIV与阿片类药物之间的相互作用。但是，我们的差距很大 了解OUD如何影响艾滋病毒感染大脑对人类艺术的细胞反应和病毒持久性的知识 或相关的模型organsims。该行政补充提案旨在生成新的地形 来自父母奖的数据集和跨海马区域的单细胞分辨率的证据，这是一个大脑区域 以非人类灵长类动物（NHP）中的HIV持久性和OUD的偏见而闻名。这些数据将提供 多种方式的前所未有的蜂窝景观可以利用，以制定限制病毒的策略 持续性，恢复并保留艾滋病毒患者的最佳大脑健康。在我们出版的初步 我们开发了创新的单细胞方法，包括单细胞同工型RNA测序 （Scisor-seq），它可以使数千个聚腺苷酸化RNA的单细胞长阅读RNA测序 单细胞。在这些新颖的测序和计算方法中，与Scatac-Seq一起 染色质的可及性，将允许映射细胞基因表达，开放染色质区域，同工型 以及跨海马单细胞检测SIV。最近的文献支持艾滋病毒的存在 大脑，更具体地说是海马内存在的小胶质细胞和星形胶质细胞。重要的是，这个 大脑区域还参与了OUD的关联学习过程。而且，我们先前在啮齿动物的研究 海马通过建立区域和细胞特异性的研究为拟议研究奠定了基础 阿片类肽和受体以及相关信号分子的分布以及这些分布如何 受性别，压力和与阿片类药物相关的学习影响。使用新的尖端单细胞Mulitomic 综合基因组学，我们在此补充应用中提出，以生成多维单细胞 数据集并在受控的SIV感染的NHP模型中定义此类观测值的相关性， 不同的阿片类药物的记录的慢性吗啡暴露，以与父母研究进行比较 羟考酮。我们还建议使用使用选择基因的单细胞表达中观察到的任何变化 原位杂交。我们建议在以下特定目标下实现这些目标：目标1。 在受控的SIV感染环境中，海马中全基因组单细胞转录组的 NHP对ART进行监测并暴露于慢性吗啡。目标2。建立全基因组的单细胞 在AIM 1中检查的组中海马的表观基因组谱。