Mechanisms of HIV Tat regulation of macrophage gene expression in neuroAIDS

HIV Tat 调节神经艾滋病巨噬细胞基因表达的机制

• 批准号: 8728413
• 负责人: Joan Weinberger Berman
• 金额: $ 25.05万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8728413
• 关键词: AIDS neuropathy; Address; Adenoviruses; Amino Acids; Apoptosis; Binding; Binding Sites; Biological Assay; Brain; CCL2 gene; CCR5 gene; CXCR4 gene; Cell Line; Cells; Central Nervous System Diseases; ChIP-seq; Chemotactic Factors; Chronic; Complex; Consequences of HIV; Cysteine; DNA; DNA Binding; Data; Development; Disease; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genome; HIV; HIV Infections; HIV tat Protein; Human; Impairment; Infection; Inflammation; Intervention; Lead; Life; Life Cycle Stages; Luciferases; Mediating; Mediator of activation protein; Microglia; Modeling; Molecular; Mutation; Neurocognitive; Neurons; Pathogenesis; Pathway Analysis; Peripheral; Population; Protein Binding; Proteins; Recruitment Activity; Regulation; Role; Site; Subfamily lentivirinae; T-Lymphocyte; Therapeutic; Therapeutic Intervention; Tissues; Trans-Activators; Viral; Viral Proteins; Virus; Virus Diseases; antiretroviral therapy; brain tissue; chemokine; cytokine; innovation; macrophage; monocyte; neuroinflammation; neurotoxic; promoter; public health relevance

DESCRIPTION (provided by applicant): HIV enters the CNS within 2 weeks after peripheral infection and viral presence persists despite cART. CNS HIV infection results in HIV Associated Neurocognitive Disorders, or HAND, in greater than 50% of the infected population. HAND is mediated, at least in part, by ongoing neuroinflammation and low level viral persistence that causes CNS damage. A major mechanism by which virus enters the brain is by the transmigration of HIV-infected monocytes across the BBB. These monocytes differentiate into macrophages in the CNS parenchyma. They elaborate neurotoxic and chemotactic factors that recruit additional infected and uninfected monocytes into the CNS, as well as virus that infects perivascular macrophages and microglia. Thus, chronic inflammation persists within the CNS. NeuroAIDS is characterized by neuronal damage and loss, but HIV does not infect neurons. Therefore, the altered function and damage seen in neurons must be due to indirect effects of HIV infection, primarily of macrophages and microglia as the major targets of infection, including the elaboration of the neurotoxic viral protein, tat. Tat is the transactivator of the virus and isstill produced despite cART. Tat also interacts with host genes to alter their expression. We and others demonstrated that tat induced cytokine secretion in macrophages, including CCL2, that mediates neuroinflammation and is highly expressed in the CNS of HIV infected people with HAND. Tat also increased the expression of CCR5 and CXCR4 on macrophages. Thus, we hypothesize that the HIV tat protein interacts with human macrophage host genes, altering their expression, resulting in macrophage dysfunction, neuroinflammation, CNS damage and subsequent development of HAND. To address this hypothesis we will use data we obtained from ChIP-seq assays that identified specific tat binding sites in the macrophage host genome and from Ingenuity Pathway Analysis (IPA) of these 67 sites. We propose three Aims. Aim 1 is to confirm and characterize the genes identified by ChIP-seq in THP-1-Tat-Flag cell lines. Aim 2 is to demonstrate alterations in expression of the genes confirmed in AIM 1 in primary human macrophages infected with adenovirus containing Tat-Flag, and with HIV, as compared to uninfected macrophages, and in tissue sections of brains obtained from HIV infected people with and without HAND. Aim 3 is to demonstrate the amino acids of tat that mediate host gene expression. Our approaches will provide new information about how tat alters host gene expression and the contribution of these dysregulated genes to the pathogenesis of HAND. It is anticipated that these data will indicate innovative strategies for therapeutic intervention to limt the consequences of HIV CNS infection.

描述（由申请人提供）：HIV在周围感染后2周内进入中枢神经系统，尽管购物车仍持续存在。中枢神经系统HIV感染导致艾滋病毒相关的神经认知障碍，或者在超过50％的感染人群中手动。至少部分是通过持续的神经炎症和低水平的病毒持久性介导的手，从而介导了手。病毒进入大脑的主要机制是通过在BBB跨BBB的HIV感染单核细胞的转移。这些单核细胞分化为CNS实质中的巨噬细胞。它们详细介绍了神经毒性和趋化因子，这些因素招募了额外的感染和未感染的单核细胞，以及感染周围巨噬细胞和小胶质细胞的病毒。因此，慢性炎症持续存在于中枢神经系统内。神经剂的特征是神经元损伤和丧失，但HIV不感染神经元。因此，神经元中看到的功能和损害的改变必须是由于HIV感染的间接作用，主要是巨噬细胞和小胶质细胞作为主要感染的主要靶标，包括阐述神经毒性病毒蛋白，TAT。 TAT是病毒的反式激活剂，尽管卡车仍会产生。 Tat还与宿主基因相互作用以改变其表达。我们和其他人表明，TAT在包括CCL2在内的巨噬细胞中诱导了细胞因子分泌，这些巨噬细胞介导了神经炎症，并且在受HIV感染的人的中枢神经系统中高度表达。 TAT还增加了CCR5和CXCR4在巨噬细胞上的表达。因此，我们假设HIV TAT蛋白与人类巨噬细胞宿主基因相互作用，改变其表达，导致巨噬细胞功能障碍，神经炎症，CNS损伤和随后的手发展。为了解决这一假设，我们将使用我们从CHIP-SEQ分析中获得的数据，这些数据确定了巨噬细胞宿主基因组中的特定TAT结合位点以及从这67个地点的Ingenuity途径分析（IPA）中获得的数据。我们提出了三个目标。 AIM 1是确认并表征THP-1-TAT-FLAG细胞系中ChIP-Seq鉴定的基因。 AIM 2是证明与未感染的巨噬细胞相比，与未感染的巨噬细胞相比，与含有TAT-FLAG的腺病毒感染的原代人巨噬细胞中确认的基因的表达改变，以及HIV，以及在没有手没有手的HIV感染者中获得的大脑。 AIM 3是证明介导宿主基因表达的TAT的氨基酸。我们的方法将提供有关TAT如何改变宿主基因表达以及这些失调基因对手发病机理的贡献的新信息。预计这些数据将表明治疗干预的创新策略，以影响HIV CNS感染的后果。