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

基本信息

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
项目类别：
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

项目摘要

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、艾滋病毒整合和大脑潜伏期甲基苯丙胺（Meth），一种艾滋病毒感染者（PWH）使用的兴奋剂成瘾的途径，加剧大脑中艾滋病毒的影响，携带艾滋病毒的储存细胞在大脑中整合原病毒挑战治疗策略并导致神经系统后果永久化，尽管），影响炎症和抗逆转录病毒治疗（ART）。与成瘾相关的神经递质如多巴胺 (DA) 调节 HIV 表达 DA 受体的靶标，包括小胶质细胞多样化群体。 HIV-1 中的染色质组织集成是理解感染和潜伏期的重要先决条件。然而，有一个在理解染色质组织和原病毒整合之间的关系方面存在关键差距 HIV-1 在小胶质细胞储存库中持续存在，特别是在最好的实验模型 SIV- 恒河猴。此外，冰毒如何改变这些关系尚不清楚。我们假设冰毒影响染色质，增强 HIV/SIV 脑靶细胞中病毒整合的敏感性，且效果各不相同通过多巴胺能投射不同的大脑区域。实现这一目标的关键是所反映的模型的进展分阶段进行，首先在体外使用人类小胶质细胞系和 iPSC 衍生的细胞系，其中 HIV 的特征整合已被定义，并且可以控制 Meth 或 DA 的影响。接下来是离体模型并在 SIV 恒河猴体内复制 HIV、慢性冰毒和 ART 抑制。在R61阶段，我们将重点关注染色质可及性和结构蛋白 CTCF 是否与 LEDGF/p75 束缚 HIV-1 整合酶，介导插入到拓扑相关的基因组边界域 (TAD)，由 Meth 或 DA 修饰。我们将评估集成模式对完整性的影响原病毒基因组，以了解功能性病毒库的实际大小。目标 1) 描绘小胶质细胞对甲基苯丙胺和插入前病毒基因组的表观基因组变化之间的关系已建立的区域明显存在 CTCF 和 H3K36me3（TAD 边界），目标 2）连接表观遗传图谱和 HIV-1 插入模式与细胞和病毒转录图谱，同时确认 SIV系统中的这些规则以及目标3）决定了DA的贡献。 Go-No-Go：如果表观基因组发生变化由Meth或DA引起的与整合位点相关，所有模型中都有类似的规则，R33阶段将转移到体内 SIV 模型，测试长期使用冰毒对 SIV 整合表观基因组脆弱性的影响小胶质细胞，与中脑边缘区域的子集异质性相关，其中 DA 投影比较丰富来控制大脑的区域。目标 4) 测试 SIV 集成站点可用性是否遵循以下规则染色质状态、可及性和 CTCF 结合可用于预测敏感性方法。目标 5) 测试多巴胺能不同的大脑区域的整合敏感性和模式是否有所不同预测和异质小胶质细胞群体。该项目能够对小胶质细胞染色质进行新颖的研究以及 SIV 在因功能、细胞密度和神经递质而不同的大脑区域中的整合动态。