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（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，轻度损伤持续存在，并可能随着长期治疗而增加，而治疗失败的受试者会出现严重的神经认知障碍，包括HAD。 脑组织在感染的早期就被定殖。然而，在无症状期，前病毒DNA很难在脑中检测到或检测不到。无论如何，长寿的病毒储库在巨噬细胞、小胶质细胞和星形胶质细胞中建立，如果实现有效的治疗，则需要根除每一种病毒。 目前的一种假设是，大脑是在疾病的复发， 单核细胞通过血脑屏障的迁移越来越多。虽然这个模型是有吸引力的，但支持数据是有限的，也有可能是未感染的单核细胞进入大脑后期分化成巨噬细胞，并扩增高度巨噬细胞嗜性变体长期存在。许多报告也描述了星形胶质细胞感染，然而，有些仍然不相信。此外，星形胶质细胞不表达CD4，感染机制尚不清楚，而该储库中存在的病毒仍不确定。 我们将调查 大脑内外的病毒库，携带HIV-1变异体，与患有HAD或正常神经系统的AIDS患者大脑中的高度巨嗜性变异体相关。我们提出三个目标：目标1：为了研究将HIV-1播种到大脑中的水库，我们将研究骨髓和血液单核细胞是否在晚期疾病中将HIV-1播种到大脑中，或者未感染的单核细胞的流入是否会扩大已经存在于大脑中的病毒。目标二：为了表征与胶质前体细胞和星形胶质细胞相关的HIV-1储库，与巨噬细胞：小胶质细胞相比，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HIV-1在巨噬细胞/小胶质细胞，胶质前体细胞和星形胶质细胞群体中的分布。目的3：通过深度测序追踪组织储库中的脑env基序和相关变体。 我们将使用深度测序方法来测量大脑及其亲属中HIV-1变体的组织分布和患病率，重点关注大脑门户。 总之，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。我们的数据将提供深入了解的门户网站，种子艾滋病毒-1进出 的大脑，并将提供一个全面的组织分布和准种，建立在脑组织中的艾滋病毒的主要水库的流行情况。我们的研究代表了对大脑内外HIV-1储库的基础研究，并将与开发新的根除方法以从脑组织中消除HIV-1相关。