Methamphetamine, HIV integration and latency in the brain

甲基苯丙胺、艾滋病毒在大脑中的整合和潜伏 甲基苯丙胺(Meth)，艾滋病毒(PWH)患者使用的兴奋剂 成瘾的途径，加重艾滋病毒在大脑中的影响，在那里携带艾滋病毒的储存细胞整合 前病毒挑战治疗策略，并有助于延续神经后果，尽管 )，影响炎症和 抗逆转录病毒治疗(ART)。 与成瘾有关的神经递质，如多巴胺(DA)，可调节HIV 表达DA受体的靶点，包括多样化的小胶质细胞群体。HIV-1中的染色质组织 集成是了解感染和潜伏期的重要前提。然而，有一个 在理解染色质组织和相关的前病毒整合之间的关系方面存在严重差距 随着HIV-1病毒在小胶质细胞储存库中的持续存在，特别是在最佳实验模型中，SIV- 恒河猴。此外，目前还不清楚Meth是如何修改这些关系的。我们假设梅斯 染色质增强HIV/SIV脑靶细胞中病毒整合敏感性的影响，并且这种影响是不同的 通过不同多巴胺能投射的大脑区域。 对这一目标至关重要的是所反映的模型的进展 分阶段，首先采用体外培养的人小胶质细胞系和IPSC来源，其中HIV的特性 整合已经被定义，并且甲基苯丙胺或DA的影响是可以控制的。其次是体外模型 并在体内SIV恒河猴体内复制HIV、慢性Meth和ART抑制。在R61阶段， 我们将专注于染色质的可及性和与之相互作用的建筑蛋白CTCF LEDGF/p75连接HIV-1整合酶，介导插入到拓扑相关基因的基因组边界 域(TADS)，由Meth或DA修改。我们将评估集成模式对 前病毒基因组以获得对功能病毒库的真实大小的洞察。目标1)描述 甲基小胶质细胞表观基因组变化与前病毒基因组插入的关系 明显存在CTCF和H3K36me3(TAD边界)的已建立区域，目标2)连接 表观遗传学图谱和HIV-1插入模式与细胞和病毒转录图谱的关系，同时确认 SIV系统中的这些规则和目标3)决定了发展议程的贡献。去-不-去：如果表观基因组发生变化 由Meth或DA引起的都链接到整合站点，在所有模型中都有类似的规则，R33阶段将转移到 体内SIV模型，以测试长期使用Meth对SIV整合表观基因组易感性的影响 小胶质细胞与中边缘DA投射丰富的亚群的异质性相关 来控制大脑的各个区域。AIM 4)测试SIV集成站点的可用性是否遵循 染色质状态、可及性和CTCF结合可用于预测在以下情况下的易感性 冰毒。目的：测试不同多巴胺能区的整合敏感性和整合模式是否有所不同 投射和异质的小胶质细胞群。该项目使对小胶质细胞染色质的新研究成为可能。 以及不同功能、细胞密度和神经递质的大脑区域的SIV整合动力学。

Methamphetamine, HIV integration and latency in the brain Methamphetamine (Meth), a stimulant drug used by people with HIV (PWH pathways in addiction, aggravating effects of HIV in the brain, where reservoir cells bearing HIV integrated provirus challenge cure strategies and contribute to perpetuating neurological consequences, despite ), influences inflammation and antiretroviral treatments (ART). Neurotransmitters involved in addiction such as dopamine (DA) modulate HIV targets that express DA receptors, including the microglia diverse population. Chromatin organization in HIV-1 integration represents an important prerequisite for understanding infection and latency. However, there is a critical gap in understanding relationships between chromatin organization and proviral integration associated with HIV-1 persistence in microglia cellular reservoirs, and particularly in the best experimental model, the SIV- rhesus macaque. Moreover, it is unknown how Meth modifies these relationships. We hypothesize that Meth impacts chromatin enhancing viral integration susceptibilities in HIV/SIV brain target cells, and that effects vary by brain region that differ in dopaminergic projections. Critical to this goal is the progression of models reflected in phases, first in vitro using human microglia cell lines and iPSC-derived, where the characteristics of HIV integration have been defined, and effects of Meth, or DA, can be controlled. This is followed by models ex vivo and in vivo in the SIV-rhesus macaques, to replicate HIV, chronic Meth and ART-suppression. In the R61 phase, we will concentrate on whether chromatin accessibility and the architectural protein CTCF, which interacts with LEDGF/p75 to tether the HIV-1 Integrase, mediate insertions into genomic boundaries of topologically associated domains (TADs), modified by Meth, or DA. We will assess the impact of integration patterns on the integrity of proviral genomes to gain insights about the real size of the functional viral reservoir. Aim 1) delineates relationships between epigenomic changes in microglia upon Meth and proviral genomes inserted into the pre- established regions with marked presence of CTCF and H3K36me3 (TAD boundaries), Aim 2) connects epigenetic profiles and HIV-1 insertion patterns with the cellular and viral transcription profiles, while confirming these rules in the SIV system, and Aim 3) determines the contribution of DA. Go-No-Go: If epigenomic changes caused by Meth or DA are linked to integration sites, with similar rules in all models, the R33 phase will move to the in vivo SIV model, to test effects of Meth chronic use on epigenomic vulnerabilities to SIV integration in microglia, in relation to subset heterogeneity in mesolimbic areas where DA projections are abundant compared to control regions of the brain. Aim 4) tests whether SIV integration site availability follows rules dictated by chromatin states, accessibility and CTCF binding that can be used to predict susceptibilities in the context of Meth. Aim 5) tests whether integration susceptibilities and patterns vary in brain areas that differ in dopaminergic projections, and heterogeneous microglia populations. This project enables novel studies on microglia chromatin and SIV integration dynamics in brain areas that differ by function, cellular density, and neurotransmitters.