Neurotropic HIV-1 Reservoirs Inside and Outside the Brain

大脑内外的神经性 HIV-1 储库

• 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
• 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

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+个体经常患有与HIV相关的神经认知疾病（手），包括HIV相关痴呆症（HAD）。在Haart时代，温和的障碍持续存在，并且可能会随着长期治疗而增加，而在治疗失败的受试者中发生了严重的神经认知障碍。 脑组织在感染早期被定殖。然而，在无症状阶段，前病毒DNA在大脑中很难检测或无法检测到。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期的病毒储层，如果实现有效的治疗方法，则需要消除每种储层。 当前的假设是，大脑在疾病中被感染后期恢复 越来越多地通过血脑屏障迁移的单核细胞。尽管该模型很有吸引力，但支持数据是有限的，并且在分化为巨噬细胞的情况下，未感染的单核细胞进入大脑，并放大了在那里长期建立的高度巨噬细胞 - 循环变体。许多报告还描述了星形胶质细胞感染，但一些报告仍然不相信。此外，星形胶质细胞不表达CD4，而感染的机制尚不清楚，而该储层中存在的病毒仍未表征。 我们将调查 在艾滋病患者的大脑中，携带与高度MAC-Tropi变体有关的HIV-1变体的大脑内外病毒储存库。我们提出了3个目标：目标1：为了研究将HIV-1种子染成大脑的储层，我们将研究骨髓和血液单核细胞在晚期疾病中是否将HIV-1纳入大脑，还是在ACT后期未感染的单核细胞的涌入，以扩大已经扩大病毒的病毒。目标2：与巨噬细胞相比，与神经胶质前体细胞和星形胶质细胞相关的HIV-1储存剂的特征：小胶质细胞相比：小胶质细胞，我们将使用磁珠富集和特定脑细胞的分类，单基因组PCR和系统发育分析来研究HIV-1在巨噬细胞/微噬细胞/微粒细胞中，毛细血管/微型前列腺和体形群体中。 AIM 3：通过深度测序跟踪组织库中的大脑ENV图案和相关变体。 我们将使用深层测序方法来测量大脑中HIV-1变体的组织分布和患病率，及其外部关注大脑门户的组织。 总而言之，我们将阐明大脑内外的组织和细胞类型，这些组织和细胞类型携带与大脑中主要的准菜相关的变体。我们的数据将提供有关播种HIV-1进入和输出的门户的见解 大脑的组织，并将提供全面的观点，可以使您在脑组织中建立HIV的主要储藏的准菜的组织分布和流行率。我们的研究代表了大脑内外HIV-1储层的基础研究，并将与开发新的根除方法相关，以消除从脑组织中消除HIV-1。