Investigating intercellular interactions between astrocytes and microglia in HIV infection and latency

研究 HIV 感染和潜伏期星形胶质细胞和小胶质细胞之间的细胞间相互作用

• 批准号: 10754844
• 负责人: James Gesualdi
• 金额: $ 3.51万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10754844
• 关键词: Acquired Immunodeficiency Syndrome; Antibodies; Astrocytes; Biological Assay; Blood Vessels; Brain; CD4 Positive T Lymphocytes; Cell Communication; Cells; Central Nervous System; Coculture Techniques; Cognitive deficits; DNA; Data; Exposure to; Frequencies; Functional disorder; Genetic Transcription; HIV; HIV Genome; HIV Infections; HIV-associated neurocognitive disorder; Human; Immune response; Incidence; Infection; Inflammation; Inflammatory; Interleukin-1 beta; Label; Learning; Macrophage; Measures; Memory; Messenger RNA; Microglia; Motor; Neuronal Dysfunction; Neuronal Injury; Oligonucleotides; PHEMX gene; Patients; Peripheral; Persons; Pharmaceutical Preparations; Population; Predisposition; Prevalence; Production; Productivity; Proteins; Publishing; RNA; Role; Site; Testing; Tissues; Viral; Viral Genome; Viral Proteins; Viral reservoir; Virus Integration; Virus Latency; Virus Replication; antibody conjugate; antiretroviral therapy; cell type; cellular targeting; chemokine; comparative; cytokine; data integration; human model; immune modulating agents; in vitro Model; in vivo; induced pluripotent stem cell; innovation; latent HIV reservoir; latent infection; neuroinflammation; response; single-cell RNA sequencing; transcriptome; transcriptomics

HIV currently infects over 38 million people worldwide. Despite the high efficacy of antiretroviral therapies, HIV persists via transcriptionally silent latent infection and long-lived viral reservoirs in tissue sites such as the central nervous system. Viral persistence in the central nervous system leads to sustained neuroinflammation, which in turn causes neuronal injury. As a result, a spectrum of deficits in memory, learning, and/or motor functions referred to as HIV-associated neurocognitive disorders can be observed in 40- 50% of people living with HIV. The largest population of central nervous system resident cells that are susceptible to HIV infection is microglia, the resident macrophages of the brain. Astrocytes are also impacted by HIV infection both through abortive integration of the viral genome and through indirect activation by proinflammatory cytokines produced by infected cells. Because microglia are challenging to model in vitro and rarely obtainable from HIV patients, the dynamics of HIV infection in microglia as well as the immune response of both microglia and astrocytes to HIV infection remain poorly understood. This proposal seeks to deepen our understanding of the neuroinflammatory response to HIV infection using IPSC models of human microglia (iMg) and astrocytes (iAst). Recent studies have shown that peripheral macrophages respond to HIV infection by producing IL-1β and my preliminary data suggests that HIV infected iMg also secrete IL-1β. Additionally, coculture of iAst and iMg leads to increased HIV replication, suggesting that iAst act to augment HIV infection. I hypothesize that IL-1β produced by infected iMg induces TNFa production by iAst, which acts on iMg to increase productive HIV infection and decrease latent infection. I will test this hypothesis in two aims: (1) Define the impact of intercellular interactions between astrocytes and microglia on HIV-induced inflammation and HIV infection, and (2) define the influence of astrocytes on HIV latency in microglia using single cell transcriptomics. These studies will increase our understanding of HIV infection in the CNS and the intercellular interactions that cause neuroinflammation in HIV associated neurocognitive disorders.

目前，全世界有3800多万人感染艾滋病毒。尽管抗逆转录病毒药物的疗效很高， 治疗中，HIV通过转录沉默的潜伏感染和长寿的病毒库持续存在， 组织部位，如中枢神经系统。中枢神经系统中的病毒持续存在 导致持续的神经炎症，进而导致神经元损伤。结果一名 记忆、学习和/或运动功能方面的一系列缺陷，称为HIV相关性 在40- 50%的艾滋病毒感染者中可以观察到神经认知障碍。最大的 易受HIV感染的中枢神经系统驻留细胞群， 小胶质细胞，大脑中的巨噬细胞。星形胶质细胞也受到艾滋病毒感染的影响 既通过病毒基因组的失败整合，也通过促炎因子的间接激活， 受感染细胞产生的细胞因子。因为小胶质细胞很难建模 在体外，很少从艾滋病毒患者，艾滋病毒感染的动力学在小胶质细胞， 小胶质细胞和星形胶质细胞对HIV感染的免疫反应仍然很差 明白这项建议旨在加深我们对神经炎症的理解， 使用人小胶质细胞（iMg）和星形胶质细胞的IPSC模型对HIV感染的反应 （iAst）。最近的研究表明，外周巨噬细胞对HIV感染的反应是， 产生IL-1β，我的初步数据表明HIV感染的iMg也分泌IL-1β。 此外，iAst和iMg的共培养导致HIV复制增加，表明iAst 增加艾滋病毒感染。我假设感染的iMg产生的IL-1β诱导 iAst产生TNF α，其作用于iMg以增加生产性HIV感染， 减少潜伏感染。我将从两个目标来检验这个假设：（1）定义 星形胶质细胞和小胶质细胞在HIV诱导的炎症中的细胞间相互作用， HIV感染，和（2）定义星形胶质细胞对HIV在小胶质细胞中潜伏期的影响，使用单一的 细胞转录组学这些研究将增加我们对中枢神经系统HIV感染的了解 以及在HIV相关的神经认知系统中引起神经炎症的细胞间相互作用 紊乱