An unbiased OMICs approach to identify mechanisms of Cocaine regulation of the HIV reservoir

一种公正的 OMIC 方法来确定可卡因调节 HIV 储存库的机制

• 批准号: 10321500
• 负责人: Rafick Pierre Sekaly
• 金额: $ 74.39万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-12-23 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321500
• 关键词: unbiased; OMICs; approach; identify; mechanisms

PROJECT ABSTRACT Substance abuse is associated with a more severe course of HIV disease as shown by the poor immune reconstitution in cART treated subjects and an increased rate of co-morbidities. The mechanisms that are involved in these poor clinical outcomes are not well understood most likely a consequence of poorly- controlled confounding factors in study cohorts, lack of quantitative assessment of drug levels,, incomplete measurement of immune function and poor understanding of the mechanisms that lead to HIV persistence. We have used systems biology a n d shown that t h e balance between pro and anti- inflammatory pathways a s major drivers of HIV persistence. Disruption of this balance leads to poor immune reconstitution in HIV infected cART treated subjects and to increased frequencies of latently infected cells. Cocaine use as shown by our preliminary results accounts for further perturbations of the balance between pro and anti inflammatory effector molecules and cells which results in lower levels of immune reconstitution in these subjects and perturbations in T cell homeostasis which have led to heightened HIV persistence in other cohorts. We have assembled a multidisciplinary group that will allow us to overcome these aforementioned hurdles and will use unbiased OMICs approaches to address our major objective which is to delineate the molecular mechanisms triggered by cocaine that lead to HIV persistence. In specific aim 1 we will use three independent state of the art assays to quantity the size and cellular localization of the HIV reservoir; we will use transcriptomics on specific cell subsets and other high density readouts including flow cytometry and system serology to identify the effector cells and molecules that are associated to increased HIV reservoir size. In specific aim 2 we will apply a global and targeted proteomics approach to validate molecular pathways triggered by cocaine that lead increased HIV persistence. Metabolomics will be used to identify the role of metabolites and particularly short chain fatty acids and cholesterol on HIV persistence. In Specific Aim 3 we will validate the mechanisms identified by gene expression and proteomics which we have shown in aims 1 and 2 to be associated to the impact of cocaine on HIV persistence; we will use a novel primary cell assay of HIV latency which allows for the first time to measure latency reactivation in multiple T cells subsets. LARA will allow us to confirm in vitro that cocaine and its metabolites can mobilize the above identified pathways and monitor their impact on the establishment of HIV latency in different memory T cell subsets and on the response to Latency Reactivating Agents.

项目摘要 药物滥用与贫困人口更严重的艾滋病毒疾病有关 在手推车治疗的受试者中的免疫重建和合并症率提高。这 这些较差的临床结果涉及的机制最有可能是 研究队列中缺乏混杂因素的结果，缺乏定量评估 药物水平，对免疫功能的不完整测量以及对机制的不良理解 导致艾滋病毒的持久性。我们已经使用了系统生物学和 炎症途径是艾滋病毒持久性的主要驱动力。这种平衡的破坏导致贫穷 在艾滋病毒感染的手推车治疗的受试者中，免疫重建受试者和潜在的频率增加 感染细胞。可卡因的使用如我们的初步结果所示 Pro和抗炎性效应子分子和细胞之间的平衡，这导致较低水平 这些受试者的免疫重建和T细胞稳态的扰动，这导致了增强 艾滋病毒在其他人群中的持久性。我们组建了一个多学科小组，这将使我们能够克服 这些上述障碍，并将采用公正的OMIC方法来解决我们的主要目标 是描绘可卡因触发的分子机制，导致可卡因导致HIV持久性。在特定目标1中 我们将使用三种独立的艺术测定状态来数量艾滋病毒的大小和细胞定位量 水库；我们将在特定的单元集和其他高密度读数（包括流量）上使用转录组学 细胞仪和系统血清学识别与HIV增加有关的效应细胞和分子 储层尺寸。在特定目标2中，我们将采用全球和有针对性的蛋白质组学方法来验证 可卡因触发的分子途径增加了HIV的持久性。代谢组学将用于 确定代谢物，尤其是短链脂肪酸和胆固醇在HIV持久性中的作用。在 特定目的3我们将验证通过基因表达和蛋白质组学确定的机制 在目标1和2中显示的与可卡因对艾滋病毒持久性的影响有关；我们将使用小说 艾滋病毒潜伏期的主要细胞测定法，该测定首次测量多个t中的潜伏期重新激活 细胞子集。 Lara将使我们能够在体外确认可卡因及其代谢物可以动员上述 识别途径并监控其对不同记忆T细胞中HIV潜伏期的影响的影响 子集以及对潜伏期重新激活剂的响应。