The effects of opioid use on HIV-1 reservoir dynamics

摘要 HIV-1在抑制性抗逆转录病毒治疗(ART)期间持续存在于记忆中的CD4+T细胞，是主要的 病毒治愈的障碍。最终，无论如何，成功的艾滋病毒根除方法都需要同样有效。 疾病阶段、潜在的免疫系统功能或医疗合并症。 艾滋病毒携带者通常接触阿片类药物，无论是用于治疗慢性疼痛的处方， 阿片类药物使用障碍的处方，或作为海洛因注射。阿片类药物暴露显示出免疫调节作用 但阿片类药物对HIV-1储存库动力学的影响尚未研究，因此迫切需要 需要有针对性的机械性研究来解决这一关键的知识差距。这项建议的目的是 应用尖端实验方法精确了解阿片类药物和阿片类药物使用障碍 影响HIV-1储存库动力学和潜伏期逆转，改变我们研究和检测HIV-1的能力 在这一重要的患者群体中接近根除。这一提议的科学前提是阿片类药物 限制HIV-1转录和翻译的重新激活，这是由于以前在RNA和 蛋白质合成。 虽然根除艾滋病毒领域的重点是潜伏期逆转的转录生物标记物，但我们的 初步的核糖体图谱数据显示，HIV-1的限速步骤是翻译，而不是转录 重新激活。核糖体图谱是一种新的技术，它使用深度测序来表征核酸酶- 体内核糖体在mRNA转录本上的保护足迹。蛋白质合成可以在全基因组范围内进行监测 当与信使核糖核酸测序(mrna-seq)相结合时，它提供了一种定量的翻译方法。 效率。我们建议利用艾滋病毒和宿主基因组信息，并研究 控制使用阿片类药物患者的HIV-1潜伏期逆转。我们假设阿片类药物的使用将限制HIV-1的潜伏期 与不存在HIV-1潜伏期逆转相比，逆转和较低的HIV-1翻译效率 阿片类药物暴露。为了确定阿片类药物对体内传统HIV-1持久性标志物的影响，我们将进行 HIV感染治疗受抑受试者横断面分析及量化比较 来自5个组的传统HIV-1持久性标志物：1)积极注射阿片类药物(n=20)，2) 美沙酮治疗(n=20)，丁丙诺啡治疗(n=20)，处方阿片类药物治疗(n=20) 慢性疼痛治疗(n=20)，非阿片类药物治疗(n=40)。我们将进行体外复活实验 参与者的PBMC和上帝抵抗军的小组。LRA诱导的HIV-1Carna和上清病毒粒子水平 将对阿片类药物使用群体的产量进行比较。准确测量全基因组宿主和HIV-1 从使用阿片类药物的患者分离出的PBMC中的翻译，平行的mRNA-seq和核糖体图谱将是 已执行。最后，我们将在40名参与者中评估纵向HIV-1宿主动力学。 从主动注射阿片类药物使用过渡到丁丙诺啡治疗阿片使用障碍。

Abstract HIV-1 persists in memory CD4+ T cells during suppressive antiretroviral therapy (ART) and is the major barrier to virus cure. Ultimately, a successful HIV eradication approach will need to work equally well regardless of disease stage, underlying immune system function, or medical comorbidities. People with HIV are commonly exposed to opioids, whether prescribed for chronic pain, prescribed for opioid use disorder, or injected as heroin. Opioid exposure has demonstrated immune modulatory effects but the impact of opioids on HIV-1 reservoir dynamics has not been studied and thus there is an urgent need for focused, mechanistic studies to address this critical knowledge gap. The purpose of this proposal is to apply cutting-edge experimental approaches to understand precisely how opioids and opioid use disorders impact HIV-1 reservoir dynamics and latency reversal and transform our ability to study and test HIV-1 eradication approaches in this important patient population. The scientific premise of this proposal is that opioids limit HIV-1 transcriptional and translational reactivation due to a previously unrecognized block in RNA and protein synthesis. While the HIV eradication field has focused on transcriptional biomarkers of latency reversal, our preliminary ribosome profiling data reveals that translation, not transcription, is the rate-limiting step in HIV-1 reactivation. Ribosome profiling is a novel technique that uses deep sequencing to characterize the nuclease- protected footprints of ribosomes on mRNA transcripts in vivo. Protein synthesis can be monitored genome-wide and when combined with mRNA sequencing (mRNA-seq) provides a quantitative measure of translation efficiency. We propose to leverage HIV and host genomic information and investigate the mechanisms that control HIV-1 latency reversal in patients using opioids. We hypothesize that opioid use will limit HIV-1 latency reversal and lower HIV-1 translation efficiencies, when compared to HIV-1 latency reversal in the absence of opioid exposure. To define the effects of opioids on traditional HIV-1 persistence markers in vivo, we will perform a cross-sectional analysis among HIV-infected treated suppressed participants and quantify and compare traditional HIV-1 persistence markers from 5 groups: individuals who are 1) actively injecting opioids (n=20), 2) on methadone maintenance (n=20), 3) on buprenorphine maintenance (n=20), 4) on prescribed opioids for chronic pain treatment (n=20), and 5) on no opioids (n=40). We will perform ex vivo reactivation experiments with participants' PBMC and a panel of LRA. LRA-induced levels of HIV-1 caRNA and supernatant virion production will be compared across opioid use groups. To accurately measure genome-wide host and HIV-1 translation in PBMC isolated from patients who use opioids, parallel mRNA-seq and ribosome profiling will be performed. Finally, we will assess longitudinal HIV-1 reservoir dynamics in a group of 40 participants during the transition from active injection opioid use to buprenorphine for opioid use disorder.