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

• 批准号: 10644020
• 负责人: Christine Cheng
• 金额: $ 152.72万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 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/10644020
• 关键词: 3-Dimensional; Acquired Immunodeficiency Syndrome; Acute; Alcohols; Astrocytes; Behavior; Biological Assay; Blood; Blood - brain barrier anatomy; Blood Vessels; Brain; Brain region; Cell Nucleus; Cells; Central Nervous System; 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; 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; Substance Use Disorder; Technology; Transcript; Validation; Variant; Vascularization; Viral Load result; Viral reservoir; acute infection; addiction; antiretroviral therapy; 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; immune cell infiltrate; in vivo; knock-down; nano-string; neurocognitive disorder; neurotoxicity; new technology; novel; overexpression; resilience; response; seroconversion; single nucleus RNA-sequencing; stimulant use; transcriptome; transcriptomics

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.

项目摘要 艾滋病毒感染者的全身免疫激活被假设为导致 慢性炎症性疾病，包括HIV相关的神经认知障碍（HAND）。急性HIV感染 在中枢神经系统中，被认为启动了一系列促炎事件，导致炎症诱导的 神经元损伤和相关的神经认知障碍，即使在本发明的组合中也是明显的 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）， 酒精已经显示出破坏BBB完整性。血脑屏障破坏可能会增加免疫细胞浸润到 中枢神经系统和促进神经胶质活化，增加炎症和神经毒性。有趣的是，渗透性增加 BBB的变化与HIV神经功能障碍的进展有关。因此， 可卡因使用和HIV感染可导致BBB破坏的叠加效应，并进一步影响HIV相关的 神经认知障碍然而，没有太多的全基因组分子水平的研究已经完成， 了解物质使用障碍和HIV感染/HAND中的BBB完整性。拟定的研究将 解决这个重要问题。我们的中心假设是可卡因滥用加剧了艾滋病毒的发病机制 通过破坏血脑屏障和脑中神经胶质细胞群失调而在CNS中引起。我们的整体 目的是在单个核水平上利用患者脑中的细胞类型特异性转录组信息 样本，以表征可卡因使用障碍对CNS神经元和神经胶质细胞、HIV感染和 手我们将描述单个核基因表达的特点，并确定失调的基因调控网络 在HIV感染个体中与可卡因滥用相关的每种神经元和神经胶质细胞群中，和/或 用手。我们也将进行计算分析，以确定神经元和神经胶质细胞的调控网络 因滥用可卡因而改变在验证和功能表征组件中， 基因在3D脑类器官模型中的表达，并将以CRISPR敲除和基因的过表达为特征。 这些目标的成功完成将通过以下方式产生重大的研究和临床影响：1）阐明如何 可卡因滥用改变了CNS中的HIV/HAND发病机制，以及2）发现候选分子来调节 可卡因滥用背景下CNS中的HIV感染或持续存在。