Effect of opioid use disorder on HIV latent reservoirs and immune dysfunction assessed by single-cell transcriptomics

项目摘要 全球约有1200万人注射毒品；其中13%是艾滋病毒携带者(PLWH)。阿片类药物 滥用是艾滋病毒感染的一条途径，也是有效抗逆转录病毒疗法(ART)的障碍。然而，它是 不清楚阿片类药物滥用是否会改变HIV的发病过程和潜伏期。延迟是一种障碍 治愈HIV，因为它导致大量受感染的静止细胞逃避抗病毒免疫 这些药物不是抗逆转录病毒治疗的靶点，并允许艾滋病毒病毒血症在治疗中断时反弹。潜伏细胞 是罕见的，缺乏可识别的生物标记物，这使得量化和表征潜伏细胞具有挑战性 水库。 我们的中心假设是，阿片类药物滥用加剧了艾滋病毒的发病和艾滋病毒的建立 通过失调宿主免疫反应而导致的潜伏期。我们的总体目标是利用转录 来自患者样本的信息，以表征阿片类药物使用障碍对宿主免疫功能的影响， 艾滋病毒复制和艾滋病毒潜伏期。我们将利用NIDA和NIAAA在圣彼得堡进行的四项现有研究。 彼得堡，俄罗斯，其参与者有很好的阿片类药物使用特点。 在R61阶段，我们将确定单细胞基因表达的特征，并确定调控失调的基因 免疫细胞中的网络与PLWH中阿片类药物滥用有关。我们还将进行计算分析 识别因阿片类药物滥用而改变的免疫细胞基因调控网络(目标1)。接下来，我们将结合单曲 细胞rna-seq和荧光原位杂交流式细胞术(Flow-FISH)鉴定转录组 在潜伏的艾滋病毒感染的细胞模型中(目标2)。 在R33阶段，我们将使用在目标2中开发和优化的工具来表征 PLWH合并和不合并阿片类药物滥用。我们还将验证免疫细胞基因网络是否被 阿片类药物滥用影响艾滋病毒转录和复制(目标3)。最后，我们将使用来自 目标1中的计算分析以测试候选分子影响HIV转录和 使用和不使用阿片类药物的PLWH之间的潜伏期。 这些目标的成功完成将通过以下方式产生重大的研究和临床影响：1)阐明 阿片类药物滥用改变了艾滋病毒的发病机制；2)产生了表征艾滋病毒潜伏期的新工具；以及3) 发现在阿片类药物滥用背景下调节艾滋病毒表达的候选分子。

Project Summary Approximately 12 million people inject drugs globally; 13% of whom are people living with HIV (PLWH). Opioid misuse is a route of HIV acquisition and a barrier to effective antiretroviral therapy (ART). However, it is unclear whether opioid misuse changes the course of HIV pathogenesis and latency. Latency is a barrier to curing HIV because it results in a reservoir of infected quiescent cells that evade the antiviral immune response, are not targeted by ART, and allow HIV viremia to rebound upon treatment interruption. Latent cells are rare and lack identifying biomarkers, making it challenging to quantify and characterize the latent cell reservoir. Our central hypothesis is that opioid misuse exacerbates HIV pathogenesis and the establishment of HIV latency by dysregulating the host immune response. Our overall objective is to exploit transcriptomic information from patient samples to characterize the effects of opioid use disorder on host immune function, HIV replication, and HIV latency. We will leverage four existing NIDA- and NIAAA-funded studies in St. Petersburg, Russia whose participants have well-characterized opioid use. In the R61 phase, we will characterize single cell gene expression and identify dysregulated gene regulatory networks in immune cells associated with opioid misuse in PLWH. We will also perform computational analysis to identify immune cell gene regulatory networks altered by opioid misuse (Aim 1). Next, we will combine single cell RNA-seq and fluorescent in situ hybridization flow cytometry (flow-FISH) to characterize the transcriptome in cell models of latent HIV infection (Aim 2). In the R33 phase, we will use tools developed and optimized in Aim 2 to characterize the latent reservoir in PLWH with and without opioid misuse. We will also validate whether immune cell gene networks altered by opioid misuse influence HIV transcription and replication (Aim 3). Lastly, we will use predictions from computational analysis in Aim 1 to test candidate molecules for their ability to influence HIV transcription and latency among PLWH with and without opioid use. Successful completion of these aims will have significant research and clinical impact by: 1) elucidating how opioid misuse alters HIV pathogenesis; 2) producing novel tools for characterizing HIV latency; and 3) discovering candidate molecules to regulate HIV expression in the context of opioid misuse.