Neurotropic HIV-1 Reservoirs Inside and Outside the Brain

大脑内外的神经性 HIV-1 储库

基本信息

批准号：
9035157
负责人：
PAUL R CLAPHAM
金额：
$ 40.58万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9035157
关键词：
AIDS Dementia Complex Acquired Immunodeficiency Syndrome Astrocytes Autopsy Basic Science Blood Blood - brain barrier anatomy Bone Marrow Brain Cells DNA Data Development Disease Drug or chemical Tissue Distribution Genome HIV HIV-1 HIV-associated neurocognitive disorder Health Highly Active Antiretroviral Therapy Immune Impairment In Vitro Individual Infection Life Liquid substance Measures Microglia Modeling Monitor Neurology Patients Phase Phylogenetic Analysis Population Prevalence Reporting Research Role Seeds Sorting - Cell Movement Source Spleen Structure of choroid plexus T-Lymphocyte Testing Time Tissues Tropism Variant Viral reservoir Virus base brain cell brain tissue cell type deep sequencing experience frontal lobe in vivo insight macrophage magnetic beads mild neurocognitive impairment monocyte neurocognitive disorder neurotropic precursor cell virus tropism

项目摘要

DESCRIPTION (provided by applicant): Untreated HIV-1+ individuals frequently suffer from HIV associated neurocognitive disorders (HAND) including HIV associated dementia (HAD). In the era of HAART, milder impairments persist and may increase with long-term therapy, while severe neurocognitive disorders including HAD occur in subjects who fail therapy. Brain tissue is colonized early in infection. However, proviral DNA is hard to detect or undetectable in brain during the asymptomatic phase. Regardless, long-lived viral reservoirs become established in macrophages, microglia and astrocytes, each of which will need to be eradicated if effective cures are realized. A current hypothesis is that the brain is reseeded late in disease by infected monocytes that increasingly migrate through the blood brain barrier. While this model is attractive, supporting data is limited and it is also possible that uninfected monocytes entering the brain late on differentiate into macrophages and amplify highly macrophage-tropic variants long established there. Many reports also describe astrocyte infection, however, some remain unconvinced. In addition, astrocytes do not express CD4 and mechanisms of infection are unclear while the virus present in this reservoir remains uncharacterized. We will investigate the viral reservoirs inside and outside the brain that carry HIV-1 variants related to highly mac-tropi variants in the brains of AIDS patients with HAD or normal neurology. We propose 3 aims: Aim 1: To investigate reservoirs that seed HIV-1 into the brain We will investigate whether bone marrow and blood monocytes reseed HIV-1 into the brain in late disease or whether the influx of uninfected monocytes late on acts to expand virus already present in the brain. Aim 2: To characterize of HIV-1 reservoirs associated with glial precursor cells and astrocytes in comparison with macrophage:microglia We will use magnetic bead enrichment and sorting of specific brain cells, single genome PCR and phylogenetic analysis to investigate HIV-1 in macrophage/microglia, glial precursor and astrocyte populations. Aim 3: To track brain env motifs and related variants in tissue reservoirs by deep sequencing. We will use deep sequencing approaches to measure the tissue distribution and prevalence of HIV-1 variants in brain and their relatives outside focusing on brain portals. In summary, we will elucidate the tissues and cell types inside and outside the brain that carry variants related to the predominant quasispecies in brain. Our data will provide insights into the portals that seed HIV-1 into and out of the brain and will provide a comprehensive view of the tissue distribution and prevalence of quasispecies that establish the major reservoirs of HIV in brain tissue. Our study represents basic research on the HIV-1 reservoirs inside and outside the brain and will have relevance for the development of new eradication approaches to eliminate HIV-1 from brain tissue.

描述（由申请人提供）：未经治疗的 HIV-1+ 个体经常患有 HIV 相关神经认知障碍 (HAND)，包括 HIV 相关痴呆 (HAD)。在HAART时代，较轻微的损伤持续存在，并可能随着长期治疗而增加，而包括HAD在内的严重神经认知障碍发生在治疗失败的受试者中。脑组织在感染早期就已定植。然而，在无症状阶段，前病毒 DNA 在大脑中很难检测到或检测不到。无论如何，长寿命的病毒储存库会在巨噬细胞、小胶质细胞和星形胶质细胞中建立，如果实现有效的治疗，每一种病毒都需要被根除。目前的一个假设是，大脑在疾病晚期被感染者重新播种单核细胞越来越多地迁移通过血脑屏障。虽然这个模型很有吸引力，但支持数据有限，而且也有可能未感染的单核细胞后期进入大脑后分化为巨噬细胞，并放大在那里长期存在的高度嗜巨噬细胞的变体。许多报告也描述了星形胶质细胞感染，但有些报告仍不相信。此外，星形胶质细胞不表达 CD4，感染机制尚不清楚，而该病毒库中存在的病毒仍不清楚。我们将调查大脑内外的病毒储存库携带 HIV-1 变异体，这些变异体与患有 HAD 或神经系统正常的艾滋病患者大脑中的高度 mac-tropi 变异体相关。我们提出了 3 个目标：目标 1：研究将 HIV-1 植入大脑的储存库我们将研究在疾病晚期，骨髓和血液单核细胞是否将 HIV-1 重新植入大脑，或者晚期未感染的单核细胞的流入是否会导致疾病扩大病毒已经存在于大脑中。目标 2：与巨噬细胞：小胶质细胞相比，表征与神经胶质前体细胞和星形胶质细胞相关的 HIV-1 储存库。我们将使用磁珠富集和特定脑细胞分选、单基因组 PCR 和系统发育分析来研究巨噬细胞/中的 HIV-1小胶质细胞、神经胶质前体和星形胶质细胞群。目标 3：通过深度测序追踪组织库中的大脑环境基序和相关变异。我们将使用深度测序方法来测量大脑及其亲属中 HIV-1 变异体的组织分布和流行率，而不关注大脑入口。总之，我们将阐明大脑内外携带与大脑主要准种相关的变异的组织和细胞类型。我们的数据将提供对传播 HIV-1 的入口和出口的见解大脑的组织分布和流行率的全面视图，这些准种在脑组织中建立了艾滋病毒的主要储存库。我们的研究代表了对大脑内外 HIV-1 储存库的基础研究，并将与开发新的根除方法以消除脑组织中的 HIV-1 相关。