Synergistic Mechanisms of chronic Innate Immune Activation in Microglia by Opiates and HIV Infection

阿片类药物和 HIV 感染对小胶质细胞慢性先天免疫激活的协同机制

基本信息

批准号：
10398922
负责人：
Christine Cheng
金额：
$ 68.96万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
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：
2020-07-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10398922
关键词：
3-Dimensional Acquired Immunodeficiency Syndrome Astrocytes Autologous Autopsy Biological Models Brain CCL2 gene CXCL10 gene Cardiovascular Diseases Cell Communication Cell Culture Techniques Cell Line Cell Nucleus Cells Cellular Metabolic Process Chronic Clustered Regularly Interspaced Short Palindromic Repeats Development Exposure to Functional disorder Future Genetic Transcription HIV HIV Infections HIV antiretroviral HIV-1 HIV-associated neurocognitive disorder Human Immune Immune System Diseases Incidence Individual Infection Inflammation Inflammatory Inflammatory Response Innate Immune Response Interferons Interruption Intervention Introns Knock-out Lead Link Malignant Neoplasms Microglia Molecular Morphine Myeloid Cells Nerve Degeneration Neurocognitive Neuroglia Neurologic Dysfunctions Neuronal Injury Neurons Neuropathogenesis Nuclear Export Opiate Addiction Opioid Organoids Osteoporosis Pathway interactions Patients Peripheral Blood Mononuclear Cell Persons Pharmaceutical Preparations Population Predisposition Production Publishing RNA Risk Signal Transduction Surveys Testing Tissues Toxic effect Viral Viral Proteins Viremia Virus Virus Diseases antiretroviral therapy base cell injury cell type chronic inflammatory disease comorbidity cytokine experience immune activation in vitro Model in vivo induced pluripotent stem cell inflammatory marker injection drug use innate immune sensing macrophage mitochondrial dysfunction monocyte mu opioid receptors neuroinflammation neurotoxicity opioid abuse opioid exposure opioid injection opioid use opioid use disorder overdose death pathogen peripheral blood prevent response single-cell RNA sequencing socioeconomics substance use therapeutic development transcriptomics viral DNA

项目摘要

ABSTRACT 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). Importantly, numerous studies have demonstrated that populations living with opioid-use disorder (OUD) have a higher occurrence of comorbid HIV infection, suggesting both potential socio-economic and neurocognitive influence. This comorbidity of OUD and HIV has also been shown to increase neuroinflammation and exacerbate the progression of HIV infection and HAND. While some studies have investigated the interactive effects of OUD and HAND in few cell types in isolation, the molecular mechanisms linking the two conditions is still largely unknown, especially in primary human microglia, neurons and astrocytes. Hence, we believe that this proposal fulfills a critical unmet need by establishing primary human neuronal cell cultures and an unbiased, systematic single cell transcriptomic survey of these cultures in the context of co-exposure of opiates and HIV-1 infection. In this proposal, we will establish 3D spheroid cultures consisting of induced pluripotent stem cell (iPSC)-derived human microglia, astrocytes and neurons, derived from 10 independent donor lines established at BUMC, that will recapitulate spatial complexity of cell-to-cell interactions in the brain, and interrogate the effect of HIV infection and morphine exposure on persistent innate immune activation. We will utilize cutting-edge single cell transcriptomics analysis to identify the microglia-intrinsic pathway that promotes induction of pro-inflammatory responses upon co-exposure to HIV and opiates and the neurodegenerative pathways contributing to neuronal cell injury. Finally, information from these transcriptomic studies will be leveraged to inform CRISPR/Cas-based knock-out strategies to selectively delete key pathways in iPSC-microglia to alleviate neuroinflammation. We believe that results from these studies will inform future therapeutic development to prevent HIV and opiate- induced neuroinflammation and prevent development of HAND in HIV+ substance use population.

抽象的持续性免疫激活是HIV-1感染体内的定义特征，也是进展为结束的驱动因素舞台艾滋病。假设在艾滋病毒患者中的全身免疫激活是为了增加慢性炎症性疾病的发生率，包括与HIV相关的神经认知疾病（手）。重要的是，许多研究表明，患有阿片类药物障碍（OUD）的人群具有合并症HIV感染的发生较高，表明潜在的社会经济和神经认知能力影响。 OUD和HIV的合并症也已被证明会增加神经炎症并加剧艾滋病毒感染和手的进展。尽管一些研究调查了OUD的互动效应并分离出几种细胞类型，连接这两种条件的分子机制仍然很大程度上是未知，特别是在原发性人类小胶质细胞，神经元和星形胶质细胞中。因此，我们相信这个建议通过建立原发性人类神经元细胞培养和无偏的系统性，可以满足至关重要的需求这些培养物在阿片类药片和HIV-1感染的背景下对这些培养物进行的单细胞转录组学调查。在此提案中，我们将建立由诱导多能干细胞（IPSC）衍生的3D球体培养物人类小胶质细胞，星形胶质细胞和神经元，源自在BUMC上建立的10个独立供体线，将概括大脑中细胞间相互作用的空间复杂性，并询问HIV的作用持续的先天免疫激活感染和吗啡暴露。我们将利用尖端的单个单元转录组学分析以识别促进促炎性诱导的小胶质细胞中的途径共同暴露于HIV和阿片类药物以及有助于神经元的神经退行性途径的反应细胞损伤。最后，这些转录组研究的信息将被利用以告知CRISPR/CAS的信息在IPSC-Microglia中有选择地删除关键途径以减轻神经炎症的敲除策略。我们认为这些研究的结果将为未来的治疗发展提供信息，以防止艾滋病毒和阿片类药物诱导神经炎症并防止HIV+药物使用种群中的手发展。