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: 8414162
• 负责人: MARCO SALEMI
• 金额: $ 60.48万
• 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 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8414162
• 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 Targeting; 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.

描述（由申请人提供）：尽管引入了高活动性抗逆转录病毒治疗（HAART），但新近HIV-1感染的患者的比例在增加与HIV相关的痴呆症（HAT）的比例正在增加。目前，没有有效的治疗方法。了解推动疾病进展过程中神经毒动菌株出现的进化因素对于开发一种现实的神经辅助模型至关重要。当前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，肺和血液中选择单核细胞/巨噬细胞，并与神经发生的发作有关。将采用神经辅助的恒河猴模型来研究疾病进展过程中病毒准菜的演变，并跟踪渗透大脑的SIV感染的SIV感染的巨噬细胞子集。 24只动物将被遗传定义的病毒群感染。外周血和组织样品将随着时间的推移收集，并用于扩增病毒基因组的3.3kb片段，包括GP160，NEF和5'LTR，以及一些来自选定组织的全长基因组。我们将使用激光捕获的显微镜在早期和末期疾病中与大脑中特定的有效感染的巨噬细胞分离病毒变异。高分辨率的系统发育，种群遗传学和分子钟算法（系统动力学）将揭示与神经性病变感染的发作有关的病毒储量的遗传方面，由于与人类组织中的组织采样相关的道德问题，这些神经性病感染尚未被特征。具体目标1将通过纵向PBMC/组织巨噬细胞采样和没有CD8+ T细胞耗竭的猴子的纵向PBMC/组织巨噬细胞在感染过程中研究淋巴机和非淋巴组织中SIV的进化动力学；特定的目标2将识别涉及大脑进入的巨噬细胞子集，并充当潜在的病毒储存剂，以引起大脑感染。我们将能够鉴定出脑感染的节奏和脑感染方式和进化特征，从而导致传染性巨噬细胞 - 授予准确的准精，可用于预测和监测疾病。同样重要的是，有可能将这些发现用于开发靶向巨噬细胞和与神经病变相关的病毒的药物。总体而言，我们将汇编迄今为止各种组织的纵向SIV序列的最全面的数据库。 PI虽然没有以前的R01资金的新调查员在遗传数据的尖端分析方面具有丰富的经验，并且已经组建了一个独特且合格的跨学科团队来协助研究。