Neurotropic HIV-1 Reservoirs Inside and Outside the Brain

大脑内外的神经性 HIV-1 储库

• 批准号: 8638069
• 负责人: PAUL R CLAPHAM
• 金额: $ 40.23万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 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/8638069
• 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; Highly Active Antiretroviral Therapy; Immune; Impairment; In Vitro; Individual; Infection; Life; Liquid substance; Measures; Microglia; Modeling; Monitor; Neurocognitive; Neurology; Patients; Phase; Phylogenetic Analysis; Population; Prevalence; Relative (related person); Reporting; Research; Role; Seeds; Sorting - Cell Movement; Source; Spleen; Structure of choroid plexus; T-Lymphocyte; Testing; Time; Tissues; Tropism; Variant; Viral; Virus; base; brain cell; brain tissue; cell type; deep sequencing; experience; frontal lobe; in vivo; insight; macrophage; magnetic beads; mild neurocognitive impairment; monocyte; neurotropic; precursor cell; public health relevance; virus tropism

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(申请人提供)：未经治疗的HIV-1+患者经常患有HIV相关神经认知障碍(HAND)，包括HIV相关痴呆(HAD)。在HAART时代，较轻微的损害持续存在，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被定植。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，在巨噬细胞、小胶质细胞和星形胶质细胞中建立了长寿的病毒库，如果实现有效的治疗，这些细胞中的每一个都需要根除。目前的一种假设是，大脑在疾病后期被感染重新播种。 单核细胞越来越多地通过血脑屏障迁移。虽然这个模型很有吸引力，但支持数据有限，而且还有可能未感染的单核细胞在进入大脑后分化为巨噬细胞，并放大长期在那里建立的高度亲巨噬细胞的变体。许多报告也描述了星形胶质细胞感染，然而，一些人仍然不信服。此外，星形胶质细胞不表达CD4，感染机制尚不清楚，而该储存库中存在的病毒仍未确定其特征。我们将调查 脑内外的病毒库携带HIV-1变种，与患有HAD或正常神经学的艾滋病患者大脑中的高度趋化变种有关。我们提出了三个目标：目标1：为了研究将HIV-1播撒到脑中的储存库，我们将调查在晚期疾病中，骨髓和血液单核细胞是否将HIV-1重新播种到脑中，或者是否晚期未感染的单核细胞的流入对扩大脑中已存在的病毒起作用。目的：比较胶质前体细胞和星形胶质细胞与巨噬细胞：小胶质细胞相关的HIV-1储存库的特征。我们将利用磁珠浓缩和特定脑细胞的分选、单基因组聚合酶链式反应和系统发育分析来研究巨噬细胞/小胶质细胞、胶质前体细胞和星形胶质细胞群体中的HIV-1。目的3：通过深度测序追踪脑组织储藏库中的脑膜蛋白基序及其相关变体。我们将使用深度测序方法来测量HIV-1变异体在大脑及其外部亲属中的组织分布和流行情况，重点是大脑门户。综上所述，我们将阐明大脑内外携带与大脑中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HIV-1进入和传出的门户的洞察 并将全面了解准种的组织分布和流行情况，这些准种确立了艾滋病毒在脑组织中的主要储存库。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有关。