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

项目总结 艾滋病毒携带者(PLHIV)的长期阿片类药物使用是一个持续的公共卫生危机。相互影响 阿片类药物和HIV之间可能会加速HIV相关的免疫失调、慢性炎症和 凝血障碍，并易患艾滋病和非艾滋病相关的死亡率和发病率。从中发布的数据 体外和动物模型表明，除了与阿片受体相互作用外，阿片类药物还直接 结合和激活Toll样受体4(TLR-4)，促进肠道损伤和微生物移位，干扰 具有调节内毒素(LPS)诱导的炎症、促进HIV复制的作用。我们自己的 初步数据支持阿片类药物的使用与晚期免疫激活和调节失调有关 PLHIV患者对脂多糖的反应。我们假设阿片类药物暴露会破坏体内平衡抗炎作用 通常限制TLR-4介导的单核细胞激活以应对内毒素的miRNAs，导致慢性先天 免疫激活和功能障碍。我们认为阿片类药物的暴露将与 肠道通透性升高，HIV相关T细胞活化和衰竭，凝血障碍，以及 抗病毒能力受损。在这项计划中，我们将利用我们对纵向临床 来自PLHIV的样本，他们也患有阿片类药物使用障碍(HIV+/OUD+)，他们正在参与 IIb期独立随机临床试验(CTN-067)，以确定联合艾滋病毒的最佳治疗 感染和OUD。试验参与者将接受抗逆转录病毒治疗(ART)和为期6个月的OUD治疗。 使用1)长效阿片拮抗剂缓释纳曲酮(XR-NTX)；或2)治疗-AS- 通常(TAU)使用阿片类激动剂。根据已发表的体外和动物模型的研究结果，我们建议 这种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强(目标1)；将恢复正常的miRNAs 调节内毒素诱导的免疫激活(AIM 2)；并将促进早期病毒控制和减少 可诱导的艾滋病毒蓄水池(目标3)。为了达到这些目的，我们将使用从CTN-067试验中收集的样本 参与者(N=350)，以及无物质依赖史或 阿片类药物暴露(HIV+/OUD-；N=100)。我们将采用先进的技术，包括：超灵敏 基于电化学发光的测量血浆细胞因子的平台；下一代测序(NGS) 静息和内毒素刺激单核细胞的miRNA和mRNA；TAT/REV诱导的限制性稀释法 (Tilda)用于定量可诱导前病毒储存库的播种；以及一种新的流式细胞仪分析来定量 包含表达GAG蛋白的前病毒和新的萌芽细胞表面病毒的CD4+T细胞亚群。我们 希望找到一种治疗策略，同时改善免疫失调和 促进病毒控制；如果成功，我们的发现将为疾病管理提供一条创新的途径 在PLHIV中，无论有无OUD。

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.