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: 8213743
• 负责人: MARCO SALEMI
• 金额: $ 61.98万
• 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/8213743
• 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），但新发HIV-1感染患者发生HIV相关痴呆（HAD）的比例正在增加。目前，HAD尚无有效的治疗方法。了解在疾病进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neuroAIDS模型至关重要。当前提案的目的是定义中枢神经系统（CNS）内的病毒进化步骤，并在神经发病机制发生之前和发生相关时从骨髓、肠道、肺和血液中选择单核细胞/巨噬细胞。神经艾滋病的恒河猴模型将用于研究疾病进展过程中病毒准种的演变，并跟踪SIV感染的巨噬细胞亚群浸润大脑。24只动物将感染基因定义的病毒群。将随时间采集外周血和组织样本，并用于扩增病毒基因组的3.3kb片段，包括gp 160、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早期和晚期疾病的大脑中特定的生产性感染的巨噬细胞中分离病毒变体。高分辨率的系统发育，群体遗传学和分子时钟算法（遗传动力学）将揭示与神经病原性感染的发病有关的病毒储库的遗传方面，由于与人类组织采样相关的伦理问题，这些病毒储库尚未被表征。具体目标1将通过纵向PBMC/组织巨噬细胞采样和有和无CD 8 + T细胞耗竭的猴脑活检研究感染过程中淋巴和非淋巴组织中SIV的进化动力学;具体目标2将鉴定参与脑进入并充当脑感染潜在病毒储库的巨噬细胞亚群。我们将能够确定大脑感染的克里思和模式以及导致感染性嗜巨噬细胞准种出现的进化特征，这些准种可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与神经发病机制相关的巨噬细胞和病毒准种的药物。总的来说，我们将编译最全面的数据库的纵向SIV序列从各种组织的日期。PI虽然是一名没有R 01资助的新研究者，但在遗传数据的尖端分析方面拥有丰富的经验，并组建了一支独特而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相关痴呆的项目研究了参与脑感染的各种组织中免疫缺陷病毒的进化。使用神经AIDS的猴子模型，其模拟人类HIV感染的过程。其结果将是对艾滋病患者痴呆症发病的遗传基础的描述，从而为开发新的诊断和治疗工具铺平道路。