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治疗 (ART) 期间持续存在于记忆 CD4+ T 细胞中，是主要的病毒治愈的障碍。最终，成功的艾滋病毒根除方法需要同样有效，无论疾病阶段、潜在的免疫系统功能或医疗合并症。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、用于治疗阿片类药物使用障碍，或作为海洛因注射。阿片类药物暴露已被证明具有免疫调节作用但阿片类药物对 HIV-1 病毒库动态的影响尚未研究，因此迫切需要需要进行有针对性的机制研究来解决这一关键的知识差距。该提案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的情况影响 HIV-1 储存库动态和潜伏期逆转，并改变我们研究和测试 HIV-1 的能力这一重要患者群体的根除方法。该提案的科学前提是阿片类药物由于 RNA 中先前未被识别的阻断而限制 HIV-1 转录和翻译再激活蛋白质合成。虽然艾滋病毒根除领域的重点是潜伏期逆转的转录生物标志物，但我们的初步核糖体分析数据表明，翻译（而非转录）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体内 mRNA 转录物上核糖体的受保护足迹。可以在全基因组范围内监测蛋白质合成当与 mRNA 测序 (mRNA-seq) 结合使用时，可提供翻译的定量测量效率。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 潜伏逆转的情况相比，HIV-1 潜伏期逆转和较低的 HIV-1 翻译效率阿片类药物暴露。为了确定阿片类药物对传统 HIV-1 体内持久性标记物的影响，我们将进行对艾滋病毒感染者接受治疗的抑制参与者进行横断面分析，并进行量化和比较来自 5 组的传统 HIV-1 持久性标记：1) 积极注射阿片类药物 (n=20)、2) 的个体美沙酮维持治疗 (n=20), 3) 丁丙诺啡维持治疗 (n=20), 4) 服用处方阿片类药物慢性疼痛治疗 (n=20)，以及 5) 不使用阿片类药物 (n=40)。我们将进行离体再激活实验与参与者的 PBMC 和 LRA 小组一起。 LRA 诱导的 HIV-1 caRNA 和上清病毒颗粒水平将比较阿片类药物使用组的产量。准确测量全基因组宿主和 HIV-1 从使用阿片类药物的患者中分离出的 PBMC 的翻译、平行 mRNA-seq 和核糖体分析将执行。最后，我们将评估 40 名参与者的纵向 HIV-1 储存库动态。从主动注射阿片类药物使用过渡到丁丙诺啡治疗阿片类药物使用障碍。