Methamphetamine, HIV integration and latency in the brain

甲基苯丙胺、艾滋病毒整合和大脑潜伏期

• 批准号: 10814672
• 负责人: Maria Cecilia Garibaldi Marcondes
• 金额: $ 60.93万
• 依托单位: SAN DIEGO BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10814672
• 关键词: Address; Affect; Animals; Architecture; Area; Basal Ganglia; Binding; Brain; Brain region; CCCTC-binding factor; Cell Line; Cell model; Cells; Central Nervous System; Central Nervous System Stimulants; Cerebellum; Characteristics; Chromatin; Chromatin Structure; Chronic; Complex; Data; Dopamine; Dopamine Receptor; Drug usage; Drug user; Epigenetic Process; Exhibits; Experimental Models; Frequencies; Gene Expression Profile; Genes; Genetic Transcription; Genome; Genomic Segment; Genomics; Goals; Grant; HIV; HIV Infections; HIV-1; HIV-1 integrase; Heterogeneity; Human; In Vitro; Infection; Inflammation; Inflammatory; Intake; Link; Location; Macaca mulatta; Mediating; Methamphetamine; Microglia; Modeling; Neurologic; Neurotransmitters; Nucleus Accumbens; Pathway interactions; Pattern; Persons; Pharmaceutical Preparations; Phase; Population; Population Heterogeneity; Predisposition; Prefrontal Cortex; Provirus Integration; Publishing; Role; SIV; Site; Stimulant; Structure; Substantia nigra structure; System; Testing; Tissues; Validation; Ventral Tegmental Area; Viral; Viral Genome; Viral reservoir; Virus Integration; Work; addiction; antiretroviral therapy; comparison control; density; epigenomics; in vitro testing; in vivo; in vivo Model; induced pluripotent stem cell; insight; integration site; methamphetamine effect; methamphetamine use; neuroAIDS; nonhuman primate; novel; response; substance use; transcriptional coactivator p75

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.

甲基苯丙胺、艾滋病毒在大脑中的整合和潜伏期 甲基苯丙胺（Methane），一种HIV感染者使用的兴奋剂（PWH 成瘾途径，加重艾滋病毒在大脑中的影响，其中携带艾滋病毒的储存细胞整合 前病毒挑战治疗策略，并有助于永久的神经系统后果，尽管 ），影响炎症， 抗逆转录病毒治疗（ART）。 参与成瘾的神经递质如多巴胺（DA）调节HIV 表达DA受体的靶点，包括小胶质细胞多样性群体。HIV-1的染色质结构 集成是理解感染和潜伏期的重要先决条件。但有一个 在理解染色质组织和前病毒整合之间的关系方面存在重大差距， HIV-1在小胶质细胞库中的持久性，特别是在最好的实验模型中，SIV- 恒河猴此外，还不知道Meth如何改变这些关系。我们假设冰毒 影响染色质增强HIV/SIV脑靶细胞中的病毒整合亲合性，并且影响各不相同 多巴胺能投射不同的大脑区域。 这一目标的关键是模型的进展反映 分阶段，首先在体外使用人小胶质细胞系和iPSC衍生的，其中HIV的特征 已经定义了整合，并且可以控制Meth或DA的影响。其次是体外模型 以及在SIV-恒河猴体内，复制HIV、慢性甲氧苯并咪唑和ART抑制。在R61阶段， 我们将集中研究是否染色质可及性和建筑蛋白CTCF，它与 LEDGF/p75系链HIV-1整合酶，介导插入到拓扑相关的 域（TADs），由Meth或DA修改。我们将评估整合模式对 前病毒基因组，以了解功能性病毒库的真实的大小。目标1）界定 Meth和前病毒基因组插入前- CTCF和H3 K36 me 3显著存在的已建立区域（边界），目标2）连接 表观遗传谱和HIV-1插入模式与细胞和病毒转录谱，同时确认 SIV系统中的这些规则，以及Aim 3）确定DA的贡献。 由于所有模型的规则相似，R33阶段将移至 体内SIV模型，以测试Meth长期使用对SIV整合的表观基因组脆弱性的影响， 小胶质细胞，与中脑边缘区的亚群异质性有关，其中DA预测丰富， 来控制大脑的某些区域。目标4）测试SIV集成站点的可用性是否遵循以下规则： 染色质状态、可接近性和CTCF结合，可用于预测 Meth.目的5）测试整合能力和模式是否在多巴胺能神经递质不同的脑区中不同。 投射和异质性小胶质细胞群体。该项目使小胶质细胞染色质的新研究成为可能 以及SIV在功能、细胞密度和神经递质不同的脑区中的整合动力学。