Single Cell Transcriptomics of the Cocaine Use Disorder in the Context of HIV

HIV 背景下可卡因使用障碍的单细胞转录组学

基本信息

批准号：
10454684
负责人：
Christine Cheng
金额：
$ 156.2万
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：
2022-06-15 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10454684
关键词：
3-Dimensional Acquired Immunodeficiency Syndrome Acute Address Alcohols Astrocytes Behavior Biological Assay Blood Blood - brain barrier anatomy Blood Vessels Brain Brain region Cell Nucleus Cells Clinical Clustered Regularly Interspaced Short Palindromic Repeats Cocaine Cocaine Users Cocaine misuse Cocaine use disorder Computer Analysis Corpus striatum structure Data Set Databases Development Disease Dorsal Event Exhibits Gene Expression Genes HIV HIV Infections HIV-1 HIV-associated neurocognitive disorder Human Immune Immunosuppression In Vitro Incidence Individual Infection Inflammation Inflammatory Interneurons Knock-in Knock-out Lead Leptomeninges Methamphetamine Methods Microglia Microtubules Modeling Molecular Neuraxis Neurocognitive Neurocognitive Deficit Neuroglia Neurologic Neurologic Dysfunctions Neuronal Injury Neurons Organoids Pathogenesis Pathway interactions Patients Pattern Pericytes Persons Pharmaceutical Preparations Population Prefrontal Cortex Primary Infection Property Regulation Research Route Sampling Small Nuclear RNA Substance Use Disorder Technology Transcript Tumor-infiltrating immune cells Validation Variant Viral Load result Viral reservoir acute infection addiction antiretroviral therapy base blood-brain barrier disruption blood-brain barrier permeabilization brain cell cell type chronic inflammatory disease cocaine use gene regulatory network gene therapy genome-wide glial activation immune activation in vivo knock-down nano-string neurocognitive disorder neurotoxicity new technology novel overexpression resilience response seroconversion stimulant use transcriptome transcriptome sequencing transcriptomics

项目摘要

PROJECT SUMMARY 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). Acute HIV 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. The use of psychostimulants (such as cocaine and methamphetamine) and alcohol has been shown to disrupt BBB integrity. Disrupted BBB may increase immune cell infiltrating into the CNS and promote glial activation, increased inflammation and neurotoxicity. Interestingly, increased permeability of BBB has been implicated in the progression of HIV neurological dysfunction. Thus, the combined effect of cocaine usage and HIV infection can cause an additive effect on BBB disruption and further impact HIV-related neurocognitive impairments. However, not much genome-wide molecular level study has been done in understanding BBB integrity in substance use disorder and in HIV infection/HAND. The proposed study will address this important question. Our central hypothesis is that cocaine misuse exacerbates HIV pathogenesis in the CNS by disrupting blood-brain barrier and 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 cocaine 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 cocaine 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 cocaine misuse. In the validation and functional characterization component, we will characterize top genes in 3D brain organoid model and will characterize with CRISPR knockout and overexpression of the gene. Successful completion of these aims will have significant research and clinical impact by 1) elucidating how cocaine 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 cocaine misuse.

项目概要据推测，艾滋病毒感染者的全身免疫激活是导致艾滋病发病率较高的原因。慢性炎症性疾病，包括 HIV 相关神经认知障碍 (HAND)。急性艾滋病毒感染中枢神经系统中的细胞被认为会引发一系列促炎症事件，从而导致炎症诱发即使在目前的组合中，神经元损伤和相关的神经认知障碍也是明显的抗逆转录病毒治疗（cART）时代。使用精神兴奋剂（如可卡因和甲基苯丙胺）和酒精已被证明会破坏血脑屏障的完整性。血脑屏障破坏可能会增加免疫细胞浸润促进中枢神经系统神经胶质细胞活化，增加炎症和神经毒性。有趣的是，渗透性增加 BBB 与 HIV 神经功能障碍的进展有关。因此，综合效应可卡因使用和 HIV 感染会对 BBB 破坏产生叠加效应，并进一步影响 HIV 相关疾病神经认知障碍。然而，目前在全基因组分子水平上进行的研究并不多。了解物质使用障碍和 HIV 感染/HAND 中的 BBB 完整性。拟议的研究将解决这个重要问题。我们的中心假设是可卡因滥用会加剧艾滋病毒的发病机制通过破坏血脑屏障和失调大脑中的神经胶质细胞群来影响中枢神经系统。我们的整体目的是利用患者大脑中单核水平的细胞类型特异性转录组信息样本来表征可卡因使用障碍对中枢神经系统神经元和神经胶质细胞、艾滋病毒感染和手。我们将表征单核基因表达并识别失调的基因调控网络在与艾滋病毒感染者滥用可卡因相关的每个神经元和神经胶质细胞群中和/或用双手。我们还将进行计算分析来识别神经元和神经胶质细胞调节网络因滥用可卡因而改变。在验证和功能表征部分，我们将表征顶部 3D 大脑类器官模型中的基因，并将通过 CRISPR 敲除和基因过度表达来表征。成功完成这些目标将通过以下方式产生重大研究和临床影响：1）阐明如何可卡因滥用会改变中枢神经系统中的 HIV/HAND 发病机制，2) 发现候选分子来调节在滥用可卡因的情况下，艾滋病毒感染或中枢神经系统持续存在。