Viral evolution in peropheral macrophages and brain during progression to AIDS

艾滋病进展过程中外周巨噬细胞和大脑的病毒进化

• 批准号: 8034217
• 负责人: MARCO SALEMI
• 金额: $ 70.85万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-15 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8034217
• 关键词: AIDS Dementia Complex; AIDS neuropathy; Acquired Immunodeficiency Syndrome; Algorithms; Amino Acids; Animal Model; Animals; Anti-Retroviral Agents; Automobile Driving; Autopsy; Biopsy; Blood; Blood specimen; Bone Marrow; Brain; Brain region; CD8B1 gene; Cells; Cerebrospinal Fluid; Data; Databases; Dementia; Developed Countries; Development; Disease; Disease Progression; Drug Delivery Systems; Epidemic; Ethics; Event; Evolution; Funding; Genes; Genetic; Genetic Programming; Genetic Recombination; Genome; Goals; HIV; HIV Infections; HIV-1; Health; Host resistance; Human; Immune; Immunologic Deficiency Syndromes; Individual; Infection; Inflammation; Lasers; Length; Link; Lung; Lymphoid; Lymphoid Tissue; Macaca mulatta; Maintenance; Mediating; Meninges; Microscopy; Mining; Modeling; Molecular; Monitor; Monkeys; Mononuclear; Neuraxis; Neuropathogenesis; Onset of illness; Pathogenesis; Patients; Pattern; Peripheral Blood Mononuclear Cell; Phagocytes; Phylogenetic Analysis; Population; Population Dynamics; Population Genetics; Population Sizes; Process; Qualifying; Research Personnel; Research Project Grants; Resolution; SIV; Sampling; Series; Staging; T-Cell Depletion; Techniques; Testing; Time; Tissue Sample; Tissues; Variant; Viral; Viral Genes; Viral Genome; Virulence; Virus; base; brain tissue; cell type; design; effective therapy; end stage disease; experience; genetic analysis; gp160; innovation; knowledge base; longitudinal database; macrophage; migration; monocyte; neurotropic; novel diagnostics; novel therapeutics; pandemic disease; peripheral blood; pressure; research study; statistics; time use; tool; treatment strategy

DESCRIPTION (provided by applicant): Despite the introduction of highly-active antiretroviral treatment (HAART), the proportion of newly HIV-1 infected patients developing HIV-associated dementia (HAD) is increasing. Currently, there is no effective therapy for HAD. Understanding the evolutionary factors driving the emergence of neurovirulent strains during disease progression is of pivotal importance to develop a realistic model of neuroAIDS. The objectives of the current proposal are to define viral evolutionary steps within the central nervous system (CNS) and select monocyte/macrophages from bone marrow, gut, lung and blood preceding and associated with the onset of neuropathogenesis. The Rhesus macaque model of neuroAIDS will be employed to study the evolution of the viral quasispecies during disease progression and to track SIV-infected macrophage subsets infiltrating the brain. 24 animals will be infected with a genetically-defined viral swarm. Peripheral blood and tissue samples will be collected over time and used for amplification of a 3.3kb fragment, including gp160, nef and 5' LTR, of the viral genome, as well as some full-length genomes from selected tissues. We will use laser-captured microscopy to isolate viral variants from specific productively infected macrophage in the brain at early and end stage disease. High-resolution phylogenetic, population genetics, and molecular clock algorithms (phylodynamics) will reveal genetic aspects of viral reservoirs linked to the onset of a neuropathogenic infection that have not yet been characterized because of ethical problems associated with tissue sampling in humans. Specific Aim 1 will investigate the evolutionary dynamics of SIV in lymphoid and non- lymphoid tissues during the course of the infection via longitudinal PBMC/tissue macrophages sampling and brain biopsies of monkeys with and without CD8+ T-cell depletion; Specific Aim 2 will identify macrophage subsets involved in brain entry and acting as potential viral reservoirs for brain infection. We will be able to identify tempo and mode of brain infection and evolutionary signatures leading to the emergence of infectious macrophage-tropic quasispecies that could be used to predict and monitor the disease. Equally important is the possibility to use the findings into developing drugs that target macrophage and viral quasispecies associated to neuropathogenesis. Overall, we will compile the most comprehensive database of longitudinal SIV sequences from a variety of tissues to date. The PI, although a new investigator without previous R01 funding, has significant experience in cutting-edge analysis of genetic data, and has assembled a unique and qualified interdisciplinary team to assist in the study. PUBLIC HEALTH RELEVANCE: This project on HIV-associated dementia examines the evolution of immunodeficiency viruses in various tissues involved in brain infection. A monkey model of neuroAIDS is used that mimics the course of HIV infection in humans. The result will be a description of the genetic basis for the onset of dementia in patients with AIDS leading the way to the development of new diagnostic and therapeutic tools.

描述(由申请人提供)：尽管引入了高效抗逆转录病毒治疗(HAART)，但新感染艾滋病毒-1的患者患上艾滋病毒相关性痴呆(HAD)的比例正在增加。目前，HAD尚无有效的治疗方法。了解在疾病发展过程中驱动神经毒力菌株出现的进化因素对于开发一个现实的神经艾滋病模型至关重要。目前的建议的目的是定义中枢神经系统(CNS)内的病毒进化步骤，并从骨髓、肠道、肺和血液中选择与神经发病之前和相关的单核/巨噬细胞。神经艾滋病的恒河猴模型将被用来研究病毒准种在疾病发展过程中的进化，并追踪SIV感染的巨噬细胞亚群渗透到大脑中。24只动物将被基因定义的病毒群感染。随着时间的推移，将收集外周血液和组织样本，用于扩增病毒基因组的3.3kb片段，包括gp160、nef和5‘Ltr，以及来自选定组织的一些全长基因组。我们将使用激光捕获显微镜从疾病早期和晚期大脑中特定的生产性感染的巨噬细胞中分离出病毒变体。高分辨率的系统发育、种群遗传学和分子时钟算法(系统动力学)将揭示与神经致病感染的发病有关的病毒库的遗传方面，由于与人类组织采样相关的伦理问题，这些方面尚未得到表征。特定目标1将通过纵向PBMC/组织巨噬细胞采样和有或没有CD8+T细胞耗尽的猴子的脑活检来研究SIV在感染过程中淋巴组织和非淋巴组织中SIV的进化动态；特定目标2将确定参与脑内进入的巨噬细胞亚群，并作为脑感染的潜在病毒库。我们将能够识别大脑感染的节奏和模式，以及导致出现感染性巨噬细胞嗜性准种的进化特征，这些准种可以用于预测和监测疾病。同样重要的是，有可能利用这些发现开发针对巨噬细胞和与神经发病相关的病毒准种的药物。总体而言，我们将汇编到目前为止各种组织中最全面的纵向SIV序列数据库。尽管PI是一名新的研究员，没有以前的R01资金，但在基因数据的尖端分析方面拥有丰富的经验，并组建了一支独特的、合格的跨学科团队来协助这项研究。公共卫生相关性：这个关于HIV相关痴呆症的项目研究了与脑感染有关的各种组织中免疫缺陷病毒的进化。一种神经艾滋病的猴子模型被用来模拟人类感染艾滋病毒的过程。这一结果将描述艾滋病患者痴呆发病的遗传基础，从而导致新的诊断和治疗工具的开发。