The impact of naltrexone treatment on opioid-induced immune and viral dysregulation during HIV-infection

项目摘要 艾滋病毒感染者（PLHIV）中的慢性阿片类药物使用是一个持续的公共卫生危机。的相互作用 阿片类药物和艾滋病毒之间的相互作用可能会加速艾滋病毒相关的免疫失调，慢性炎症和 凝血病，易患艾滋病和非艾滋病相关的死亡率和发病率。发布的数据来自 体外和动物模型表明，除了与阿片受体的相互作用外，阿片直接 结合并激活Toll样受体-4（TLR-4），促进肠道损伤和微生物易位，干扰 具有调节脂多糖（LPS）诱导的炎症，并促进HIV复制的作用。我们自己 初步数据支持阿片类药物的使用与高级免疫激活和失调有关， 对LPS的反应。我们假设阿片类药物暴露破坏了体内稳态抗炎作用， 通常限制TLR-4介导的单核细胞对LPS的活化的miRNA，导致慢性先天性 免疫激活和功能障碍。我们认为阿片类药物暴露将与以下证据相关 晚期肠通透性、HIV相关T细胞活化和耗竭、凝血病，以及 抗病毒能力受损在这项提案中，我们将利用我们获得的纵向临床 来自艾滋病毒感染者的样本也患有阿片类药物使用障碍（HIV+/OUD+），他们正在参加一项 一项独立的IIb期随机临床试验（CTN-067），以确定联合HIV的最佳管理 感染和OUD。试验参与者将接受抗逆转录病毒治疗（ART）加6个月的OUD治疗 与1）长效阿片类拮抗剂延长释放纳洛酮（XR-NTX）;或2）治疗-作为 通常（TAU）与阿片类激动剂。基于已发表的体外和动物模型的研究结果，我们建议 XR-NTX将与肠道通透性、免疫激活和 衰竭、凝血障碍和T细胞抗病毒能力的增加（目的1）;将恢复正常情况下 调节LPS诱导的免疫激活（目的2）;并将促进早期病毒控制和减少 诱导型HIV储库（Aim 3）。为了实现这些目标，我们将使用从CTN-067试验中采集的样本 参与者（N=350），以及没有药物依赖史的PLHIV参考队列， 阿片类药物暴露（HIV+/OUD-; N=100）。我们将采用先进的技术，包括： 用于测量血浆细胞因子的基于电化学发光的平台; 来自静息和LPS刺激的单核细胞的miRNA和mRNA;达特/rev诱导的有限稀释试验 （TILDA）来定量诱导型前病毒库的接种;以及一种新的流式细胞术测定来定量 CD 4 + T细胞亚群含有表达gag蛋白的前病毒和新的出芽细胞表面病毒体。我们 希望确定一种OUD治疗策略，该策略同时改善免疫失调， 促进病毒控制;如果成功，我们的研究结果将为疾病管理提供创新途径 在有和没有OUD的PLHIV中。

PROJECT SUMMARY Chronic opioid use among people living with HIV (PLHIV) is an on-going public health crisis. The interplay between opioids and HIV may accelerate HIV-associated immune dysregulation, chronic inflammation and coagulopathy, and predispose to AIDS and non-AIDS related mortality and morbidities. Published data from in vitro and animal models demonstrate that, in addition to their interactions with opioid receptors, opioids directly bind and activate Toll-like Receptor-4 (TLR-4), promote intestinal damage and microbial translocation, interfere with regulation of lipopolysaccharide (LPS)-induced inflammation, and facilitate HIV replication. Our own preliminary data support that opioid use is associated with advanced immune activation and dysregulated responses to LPS among PLHIV. We hypothesize that opioid exposure disrupts homeostatic anti-inflammatory miRNAs that normally limit TLR-4 mediated monocyte activation in response to LPS, leading to chronic innate immune activation and dysfunction. We propose that opioid exposure will be associated with evidence of advanced intestinal permeability, HIV-associated T cell activation and exhaustion, coagulopathy, and compromised antiviral capacity. In this proposal, we will capitalize on our access to longitudinal clinical samples from PLHIV who also suffer from opioid-use disorders (HIV+/OUD+), who are participating in an independent phase IIb randomized clinical trial (CTN-067) to determine optimal management of combined HIV infection and OUD. Trial participants will receive antiretroviral therapy (ART) plus 6 months of OUD therapy with either 1) the long-acting opioid-antagonist extended release naltrexone (XR-NTX); or 2) treatment-as- usual (TAU) with opioid-agonists. Based on published findings from in vitro and animal models, we propose that XR-NTX will be associated with significant reductions in gut permeability, immune activation and exhaustion, coagulopathy, and gains in T cell anti-viral capacity (Aim 1); will restore miRNAs that normally regulate LPS-induced immune activation (Aim 2); and will promote early viral control and reduction of the inducible HIV reservoir (Aim 3). To address these aims, we will use samples collected from CTN-067 trial participants (N=350), as well as a reference cohort of PLHIV with no history of substance dependency or opioid exposure (HIV+/OUD-; N=100). We will employ advanced techniques including: an ultrasensitive electrochemiluminescence-based platform to measure plasma cytokines; next generation sequencing (NGS) of miRNA and mRNA from resting and LPS-stimulated monocytes; a Tat/rev Induced Limiting Dilution Assay (TILDA) to quantify seeding of the inducible provirus reservoir; and a novel flow cytometry assay to quantify CD4+ T cell subsets that contain provirus expressing the gag protein and new budding cell surface virions. We hope to identify an OUD treatment strategy that simultaneously ameliorates immune dysregulation and promotes viral control; if successful, our findings would provide an innovative pathway to disease management in PLHIV both with and without OUD.