Single Cell Transcriptomics of the Opioid Use Disorder and HIV Syndemic in the Human Brain

人脑中阿片类药物使用障碍和艾滋病毒综合症的单细胞转录组学

基本信息

批准号：
10241384
负责人：
Christine Cheng
金额：
$ 110.59万
依托单位：
J. CRAIG VENTER INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-11-05 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10241384
关键词：
3-Dimensional Acquired Immunodeficiency Syndrome Addictive Behavior Affect Amygdaloid structure Astrocytes Brain Brain region Cell Nucleus Cells Chronic Clinical Clustered Regularly Interspaced Short Palindromic Repeats Computer Analysis Corpus striatum structure Data Set Databases Development Disease Drug Addiction Drug usage Event Exhibits Exposure to Family Gene Expression Genes HIV HIV Infections HIV-1 HIV-associated neurocognitive disorder Human Immunohistochemistry Immunosuppression In Vitro Incidence Individual Infection Inflammation Inflammatory Knock-out Lead Life Link Microglia Molecular Morphine Neuroglia Neurologic Neuronal Injury Neurons Nucleus Accumbens Opioid Organoids Pathogenesis Pathology Pathway interactions Patients Pattern Pharmaceutical Preparations Phenotype Population Prefrontal Cortex Primary Infection Public Health Regulator Genes Research Route Sampling Slice Small Nuclear RNA Social Impacts Substance Use Disorder Technology Testing Tissue-Specific Gene Expression Transcript Validation Variant Viral Load result Viral reservoir acute infection addiction antiretroviral therapy base brain cell brain tissue cell type chronic inflammatory disease differential expression economic impact epigenetic regulation gene therapy immune activation in vivo induced pluripotent stem cell lead candidate neurocognitive disorder new technology novel opioid misuse opioid use disorder resilience response seroconversion three dimensional cell culture transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY Persistent immune activation is the defining feature of HIV-1 infection in vivo and a driver of progression to end-stage AIDS. Systemic immune activation in people living with HIV has been hypothesized to account for higher incidence of chronic inflammatory diseases, including HIV-associated neurocognitive disorders (HAND). While significant neurological complications associated with HIV-1 infection occur years after seroconversion and is commonly coincident with progressive immunosuppression and high viral loads, establishment of a virus reservoir in the CNS occurs with primary infection. Furthermore, acute infection in the CNS is thought to initiate a cascade of pro-inflammatory events that result in inflammation- induced neuronal injury and associated neurocognitive disorders that are evident even in the present combination antiretroviral therapy (cART) era. Approximately 12 million people inject drugs globally, 13% of these are people living with HIV. Opioid misuse is a route of HIV acquisition and a barrier to effective antiretroviral therapy (ART). However, it is unclear whether opioid misuse changes the course of HIV pathogenesis, especially on HIV-associated neurocognitive disorders. Our central hypothesis is that opioid misuse exacerbates HIV pathogenesis in the CNS by dysregulating the glial population in the brain. Our overall objective is to exploit cell type specific transcriptomic information at the single nuclei level from patient brain samples to characterize the effects of opioid use disorder on CNS neuronal and glial cells, HIV infection and HANDs. We will characterize single nuclei gene expression and identify dysregulated gene regulatory networks in each of the neuronal and glial populations associated with opioid misuse in HIV infected individuals and/or with HANDs. We will also perform computational analysis to identify neuronal and glial cell regulatory networks altered by opioid misuse. In the validation component, we will select the top 20 targets from single cell transcriptomics profiles, first validating with immunohistochemistry with fixed brain tissue, then selecting a few top targets and using 2D and 3D cultures of iPS derived neurons, microglia and astrocytes to characterize functionality with CRISPR knockout. Successful completion of these aims will have significant research and clinical impact by 1) elucidating how opioid misuse alters HIV/HAND pathogenesis in the CNS, and 2) discovering candidate molecules to regulate HIV infection or persistence in the CNS in the context of opioid misuse.

项目概要持续的免疫激活是体内 HIV-1 感染的决定性特征，也是进展的驱动因素到艾滋病末期。据推测，艾滋病毒感染者的全身免疫激活导致慢性炎症性疾病（包括与艾滋病毒相关的神经认知疾病）发病率较高疾病（手）。虽然会发生与 HIV-1 感染相关的严重神经系统并发症血清转化后数年，通常与进行性免疫抑制和高病毒载量，中枢神经系统中病毒库的建立是在初次感染时发生的。此外，急性中枢神经系统感染被认为会引发一系列促炎症事件，从而导致炎症—— 诱发的神经元损伤和相关的神经认知障碍即使在现在也很明显联合抗逆转录病毒疗法（cART）时代。全球约有 1200 万人注射毒品，占 13% 其中包括艾滋病毒感染者。阿片类药物滥用是艾滋病毒感染的一种途径，也是有效治疗的障碍抗逆转录病毒治疗（ART）。然而，目前尚不清楚阿片类药物的滥用是否会改变艾滋病毒的病程发病机制，特别是与艾滋病毒相关的神经认知障碍。我们的中心假设是阿片类药物滥用会导致大脑神经胶质细胞群失调，从而加剧中枢神经系统中的艾滋病毒发病机制。我们的总体目标是在单核水平上利用细胞类型特异性转录组信息患者大脑样本来表征阿片类药物使用障碍对中枢神经系统神经元和神经胶质细胞的影响， HIV 感染和手。我们将表征单核基因表达并识别失调每个神经元和神经胶质细胞群中的基因调控网络与阿片类药物滥用相关 HIV 感染者和/或手。我们还将进行计算分析来识别阿片类药物滥用改变了神经元和神经胶质细胞的调节网络。在验证组件中，我们将从单细胞转录组学概况中选择前 20 个目标，首先使用使用固定脑组织进行免疫组织化学，然后选择一些顶级目标并使用 2D 和 3D iPS 衍生的神经元、小胶质细胞和星形胶质细胞培养物，通过 CRISPR 表征功能昏死。成功完成这些目标将对研究和临床产生重大影响：1) 阐明阿片类药物滥用如何改变中枢神经系统中的 HIV/HAND 发病机制，以及 2) 发现候选药物在阿片类药物滥用的情况下调节艾滋病毒感染或中枢神经系统持续存在的分子。