Single Cell Transciptomics 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：
10670632
负责人：
Christine Cheng
金额：
$ 95.59万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10670632
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 Persons Pharmaceutical Preparations Phenotype Population Prefrontal Cortex Primary Infection Public Health 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 regulatory network 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 syndemic 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.

项目摘要持续的免疫激活是HIV-1感染体内的定义特征和进展的驱动力终端艾滋病。艾滋病毒患者的全身免疫激活已被认为考虑到慢性炎性疾病的发病率更高，包括与HIV相关的神经认知疾病（手）。虽然发生与HIV-1感染相关的明显神经系统并发症血清转化后几年，通常与进行性免疫抑制和高病毒载荷，在中枢神经系统中建立病毒储存库发生在原发性感染时。此外，急性中枢神经系统感染被认为会引发一系列促炎事件，导致炎症 - 诱导的神经元损伤和相关的神经认知障碍，即使在目前抗逆转录病毒疗法（CART）时代。全球大约有1200万人注入毒品，13％其中是感染艾滋病毒的人。阿片类药物滥用是收购HIV的途径，也是有效的障碍抗逆转录病毒疗法（ART）。但是，目前尚不清楚阿片类药物是否滥用改变艾滋病毒的进程发病机理，特别是在与HIV相关的神经认知障碍上。我们的中心假设是阿片类药物滥用通过使大脑中的神经胶质种群失调而加剧了中枢神经系统中的HIV发病机理。我们的总体目的是从单核水平利用细胞类型的特定转录组信息患者脑样本以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，艾滋病毒感染和手。我们将表征单核基因表达并鉴定失调与阿片类药物滥用相关的每个神经元和神经胶质种群中的基因调节网络艾滋病毒感染的人和/或用手。我们还将执行计算分析以识别阿片类药物滥用改变了神经元和神经胶质细胞调节网络。在验证组件中，我们将从单细胞转录组学轮廓中选择前20个目标，首先用与固定脑组织的免疫组织化学，然后选择一些顶部目标，然后使用2D和3D IPS衍生的神经元，小胶质细胞和星形胶质细胞的培养物，以CRISPR为特征昏死。这些目标的成功完成将产生重大的研究和临床影响1）阐明阿片类药物滥用如何改变中枢神经系统中的艾滋病毒/手发病机理，2）发现候选者在阿片类药物滥用的背景下，分子调节中枢神经系统中的艾滋病毒感染或持久性。