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）：尽管采用了高效抗逆转录病毒治疗（HAART），但新感染 HIV-1 的患者发展为 HIV 相关痴呆（HAD）的比例仍在增加。目前，HAD 尚无有效治疗方法。了解疾病进展过程中神经毒性菌株出现的进化因素对于开发神经艾滋病的现实模型至关重要。当前提案的目标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并与之相关的骨髓、肠道、肺和血液中选择单核细胞/巨噬细胞。神经艾滋病恒河猴模型将用于研究疾病进展过程中病毒准种的进化，并追踪浸润大脑的 SIV 感染巨噬细胞亚群。 24 只动物将被基因定义的病毒群感染。随着时间的推移，将收集外周血和组织样本，并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末期大脑中特定的有效感染巨噬细胞中分离出病毒变异体。高分辨率系统发育、群体遗传学和分子时钟算法（系统动力学）将揭示与神经病原性感染发作相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，这些感染尚未得到表征。具体目标 1 将通过纵向 PBMC/组织巨噬细胞取样和有或没有 CD8+ T 细胞耗竭的猴子的脑活检，研究感染过程中 SIV 在淋巴和非淋巴组织中的进化动态；具体目标 2 将识别参与大脑进入并充当脑部感染潜在病毒储存库的巨噬细胞亚群。我们将能够识别大脑感染的节奏和模式以及导致感染性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该疾病。同样重要的是，有可能利用这些发现来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，我们将编译迄今为止最全面的各种组织纵向 SIV 序列数据库。 PI 虽然是一位新研究者，之前没有获得 R01 资助，但在基因数据的前沿分析方面拥有丰富的经验，并组建了一支独特且合格的跨学科团队来协助研究。公共健康相关性：这个关于艾滋病毒相关痴呆的项目研究了免疫缺陷病毒在涉及脑部感染的各种组织中的进化。使用神经艾滋病猴模型来模拟人类艾滋病毒感染过程。其结果将描述艾滋病患者痴呆症发病的遗传基础，从而引领新诊断和治疗工具的开发。