The effects of opioid use on HIV-1 reservoir dynamics

阿片类药物使用对 HIV-1 病毒库动态的影响

• 批准号: 10620076
• 负责人: Manish Sagar
• 金额: $ 89.75万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10620076
• 关键词: Address; Biological Markers; Blood; Buprenorphine; Cells; Cross-Sectional Studies; DNA; Data; Disease; Disease Outbreaks; Exposure to; Future; Genetic Transcription; Genomics; HIV; HIV Infections; HIV-1; Heroin; Histone Deacetylase Inhibitor; Human immunodeficiency virus test; Immune; Incubated; Individual; Infection; Interleukin-2; Knowledge; Magnetism; Maintenance; Measures; Mediating; Medical; Messenger RNA; Methadone; Methods; Monitor; Morphine; Opioid; Opioid Antagonist; Opioid Receptor; Opioid agonist; Participant; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phase; Plasma; Production; Protein Biosynthesis; Proteins; Proviruses; RNA; RNA chemical synthesis; Relapse; Research; Residual state; Ribosomes; Risk; Sampling; T-Lymphocyte; Techniques; Time; Transcript; Transcription Factor AP-1; Transcription Initiation; Translations; Viral Cytopathogenic Effect; Viral Genome; Viremia; Virion; Virus; Work; antiretroviral therapy; base; buprenorphine treatment; chronic pain; chronic pain management; comorbidity; deep sequencing; design; experimental study; genome-wide; immune system function; improved; in vivo; innovative technologies; kappa opioid receptors; mRNA sequencing; memory CD4 T lymphocyte; methadone treatment; next generation; novel; nuclease; opioid epidemic; opioid exposure; opioid injection; opioid use; opioid use disorder; opioid user; overdose risk; patient population; patients who use opioids; prescription opioid; primary endpoint; receptor expression; recruit; ribosome profiling; substance use; transcription factor

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.

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性CD 4 + T细胞中，是主要的免疫缺陷病毒。 病毒治疗的障碍。最终，一个成功的艾滋病毒根除方法将需要同样有效， 疾病阶段、潜在免疫系统功能或医学合并症。 艾滋病毒感染者通常会接触阿片类药物，无论是用于治疗慢性疼痛， 用于阿片类药物使用障碍，或作为海洛因注射。阿片类药物暴露已证明免疫调节 但阿片类药物对HIV-1储库动态的影响尚未研究，因此迫切需要 需要进行重点突出的机械研究，以解决这一关键的知识差距。这项建议的目的是 应用尖端的实验方法来准确了解阿片类药物和阿片类药物使用障碍 影响HIV-1储库动态和潜伏期逆转，并改变我们研究和检测HIV-1的能力 在这一重要的患者人群中根除方法。这项提议的科学前提是， 限制HIV-1转录和翻译再活化，这是由于RNA中以前未识别的阻断， 蛋白质合成。 虽然HIV根除领域已经集中在潜伏逆转的转录生物标志物上，但我们的研究发现， 初步的核糖体分析数据显示，翻译而不是转录是HIV-1的限速步骤 重新激活核糖体分析是一种新技术，它使用深度测序来表征核酸酶- 保护核糖体在体内mRNA转录物上的足迹。蛋白质合成可以在全基因组范围内进行监测 当与mRNA测序（mRNA-seq）结合时， 效率我们建议利用艾滋病毒和宿主基因组信息，并研究其机制， 在使用阿片类药物的患者中控制HIV-1潜伏期逆转。我们假设阿片类药物的使用会限制HIV-1的潜伏期， 逆转和较低的HIV-1翻译效率，当与HIV-1潜伏期逆转的情况下， 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 在HIV感染者中进行横断面分析， 来自5组的传统HIV-1持久性标志物：1）主动注射阿片类药物的个体（n=20），2） 接受美沙酮维持治疗（n=20），3）接受丁丙诺啡维持治疗（n=20），4）接受处方阿片类药物， 慢性疼痛治疗组（n=20）; 5）无阿片类药物治疗组（n=40）。我们将进行体外再激活实验 与受试者的PBMC和一组LRA进行比较。LRA诱导的HIV-1 caRNA和上清液病毒体水平 将在阿片类药物使用组之间进行比较。准确测量全基因组宿主和HIV-1 在从使用阿片类药物的患者中分离的PBMC中，平行的mRNA-seq和核糖体谱分析将被用于 执行。最后，我们将在一组40名参与者中评估纵向HIV-1储库动态， 从主动注射阿片类药物过渡到丁丙诺啡治疗阿片类药物使用障碍。