The effects of opioid use on HIV-1 reservoir dynamics

阿片类药物的使用对 HIV-1 病毒库动态的影响

基本信息

批准号：
10673865
负责人：
Manish Sagar
金额：
$ 88.36万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
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/10673865
关键词：
Address Biological Markers Blood Buprenorphine Cell Separation 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 Proliferating Protein Biosynthesis Proteins Proviruses RNA RNA chemical synthesis Relapse Research Residual state Ribosomes Risk Sampling T-Cell Activation T-Lymphocyte Techniques Transcript Transcription Factor AP-1 Transcription Initiation Translations Viral Cytopathogenic Effect Viral Genome Viremia Virion Virus Work antiretroviral therapy 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 nuclear factors of activated T-cells 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

项目摘要

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.

抽象的 HIV-1在抑制性抗逆转录病毒疗法（ART）期间的记忆CD4+ T细胞持续存在，是主要的病毒治愈的障碍。最终，成功的消除艾滋病毒方法都需要同样好事疾病阶段，潜在的免疫系统功能或医学合并症。患有艾滋病毒的人通常暴露于阿片类药物，无论是为慢性疼痛而开的规定用于阿片类药物使用障碍或注射为海洛因。阿片类药物暴露已显示出免疫调节影响但是尚未研究阿片类药物对HIV-1储层动力学的影响，因此很紧急需要专注的机械研究来解决这一关键知识差距。该提议的目的是采用尖端的实验方法来精确了解阿片类药物和阿片类药物的使用障碍影响HIV-1储层动力学和潜伏期逆转，并改变了我们研究和测试HIV-1的能力根除在这个重要的患者人群中。该提议的科学前提是阿片类药物由于RNA和蛋白质合成。虽然艾滋病毒根除领域的重点是延迟逆转的转录生物标志物，但我们初步核糖体分析数据表明，翻译而非转录是HIV-1中的限制步骤重新激活。核糖体分析是一种新型技术，它使用深层测序来表征核酸酶 - 核糖体在体内mRNA转录本上受保护的足迹。蛋白质合成可以监测全基因组当与mRNA测序（mRNA-SEQ）结合使用时，提供了一种定量的翻译度量效率。我们建议利用艾滋病毒和托管基因组信息，并研究的机制使用阿片类药物的患者控制HIV-1潜伏期逆转。我们假设阿片类药物的使用将限制HIV-1潜伏期与没有HIV-1潜伏期逆转相比，HIV-1的逆转和较低阿片类药物暴露。为了定义阿片类药物对体内传统HIV-1持续标记的影响，我们将执行对HIV感染的受抑制参与者进行横截面分析，并量化和比较来自5组的传统HIV-1持续标记：1）积极注射阿片类药物（n = 20），2）关于美沙酮维护（n = 20），3）在丁丙诺啡维护上（n = 20），4）在规定的阿片类药物上慢性疼痛治疗（n = 20）和5）无阿片类药物（n = 40）。我们将执行离体重新激活实验与参与者的PBMC和LRA小组。 LRA诱导的HIV-1 carNA和上清液的水平将在阿片类药物使用组中进行比较。准确测量全基因组宿主和HIV-1 从使用阿片类药物，平行mRNA-SEQ和核糖体分析的患者中分离出的PBMC翻译将是执行。最后，我们将评估40名参与者的纵向HIV-1储层动力学从主动注射阿片类药物使用到丁丙诺啡作为阿片类药物使用障碍的过渡。