Therapeutic and mechanistic significance of altered metabolism of HIV medicines by alcohol- or alcohol/synthetic opioid combination

酒精或酒精/合成阿片类药物组合改变 HIV 药物代谢的治疗和机制意义

• 批准号: 10700069
• 负责人: JASON T BLACKARD
• 金额: $ 69.72万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-10 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10700069
• 关键词: AIDS prevention; Acquired Immunodeficiency Syndrome; Adherence; Alcohol abuse; Alcohol consumption; Alcohols; Anti-HIV Agents; Antiviral Response; Apoptosis; Behavior; Behavioral; Biological Markers; Carboxylesterase 1; Cells; Cessation of life; Diphosphates; Ensure; Enzymes; Epidemic; Esterification; Ethanol; Exposure to; Fentanyl; Foundations; Free Radicals; Fumarates; General Population; Goals; HIV; HIV Infections; HIV Seronegativity; HIV risk; HIV/AIDS; Health; Hepatitis Viruses; Hepatocyte; Hepatotoxicity; Human; Hybrids; Hydrolysis; In Vitro; Incubated; Individual; Injections; Intervention; Link; Medicine; Metabolism; Monitor; Ohio; Organ; Oxidative Stress; Patients; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phosphorylation; Prevalence; Prevention; Prodrugs; Regimen; Research Design; Risk; Risk Behaviors; Safety; Sampling; Technology; Tenofovir; Testing; Therapeutic; Time; Tissues; Toxic effect; Viral Load result; Viral hepatitis; alcohol use disorder; biomarker validation; carboxylesterase; chemokine; chronic liver disease; co-infection; cytotoxicity; design; drug metabolism; global health; high risk; in vivo; mortality; multidisciplinary; opioid mortality; originality; pre-exposure prophylaxis; preventive intervention; receptor; single-cell RNA sequencing; stem; substance use; synthetic opioid; transcriptome; transcriptome sequencing; transcriptomics; treatment adherence; treatment as prevention; virology

Abstract HIV/AIDS continues to be a major global health issue, and chronic liver disease has become a major cause of HIV-mortality. Several contributing factors are recognized including hepatotoxicity of anti-HIV drugs, coinfection of hepatitis viruses and widespread alcohol use. The likelihood of concurrent use of other addictive substances such as fentanyl is high. Excessive alcohol use has been linked directly to increased HIV viral load, engagement in HIV-risk behaviors, and poor adherence in HIV prevention interventions. In addition, alcohol and fentanyl are known to alter the functionality of drug metabolizing enzymes and transporters, critical determinants for the efficacy and safety of anti-HIV drugs. Indeed, we have shown that ethanol (alcohol) exposure decreased the hydrolysis of tenofovir alafenamide fumarate (TAF) and produced a new metabolite: ethyl TAF. This metabolite is a hybrid molecule with part from TAF and part from ethanol. TAF is a new version of TDF (tenofovir disoproxil fumarate) with decreased organ toxicity, and both tenofovir prodrugs are hydrolyzed and followed by phosphorylation to produce therapeutically active metabolite. TAF is hydrolyzed by carboxylesterase-1 (CES1), while TDF is hydrolyzed predominantly by CES2. TAF and TDF are listed in ~75% anti-HIV regimens used for both pre-exposure prophylaxis (PrEP) and treatment as prevention (TasP). Our Preliminary Study has also shown that fentanyl increased the expression of chemokine co- receptors and led to increased HIV infection. The central hypothesis of this project is that metabolism-based interactions of tenofovir drugs with alcohol +/- fentanyl produce signature changes that reliably indicate efficacy and safety of tenofovir regimens and serve as a foundation for developing adherence-monitoring and PrEP/TasP intervention strategies. The Specific Aims are: (1) to characterize metabolite signatures and mechanistic biomarkers, and (2) to ascertain the significance of alcohol +/- fentanyl-altered metabolism. To determine the metabolite signatures specific to alcohol +/- fentanyl, human primary hepatocytes and peripheral blood mononuclear cells (PBMCs) will be treated with a tenofovir-regimen in the presence or absence of alcohol (+/- fentanyl); the metabolite signatures will be determined by LC-MS/MS. The altered metabolite signatures will be ascertained by in vivo studies. Transcriptome analysis will be performed at both tissue (e.g., PBMC) and single cell level (selected samples) to identify mechanistic biomarkers. To establish the significance, hepatocytes and PBMCs will be incubated with a tenofovir-regimen with or without alcohol +/- fentanyl, and viral load and cytotoxicity will be monitored. The connection of metabolite signatures and biomarkers with efficacy and safety will be ascertained in vivo. Finally, PBMCs from HIV-negative patients exposed to PrEP, PrEP+alcohol or PrEP+alcohol/fentanyl will be tested against HIV infection ex vivo. The combination of in vitro, ex vivo, and in vivo study design ensures high scientific rigor.

抽象的 艾滋病毒/艾滋病仍然是全球主要的健康问题，慢性肝病已成为 HIV验尸。认识到几种促成因素，包括抗HIV药物的肝毒性， 肝炎病毒和广泛使用饮酒。同时使用其他成瘾性的可能性 这样的物质 因为芬太尼很高。过量的饮酒已直接与HIV病毒升高有关 负载，参与艾滋病毒风险行为以及艾滋病毒预防干预措施的依从性差。此外， 众所周知，酒精和芬太尼会改变药物代谢酶和转运蛋白的功能，关键 抗HIV药物有效性和安全性的决定因素。确实，我们已经表明乙醇（酒精） 暴露减少了 Tenofovir alafenamide富马酸酯（TAF），并产生了一种新的代谢物： 乙基TAF。该代谢产物是一种杂交分子，部分来自TAF，部分来自乙醇。 TAF是新的 具有降低器官毒性的TDF（Tenofovir disoproxil fumarate）的版本，这两个Tenofovir前药都是 水解并进行磷酸化以产生治疗活性代谢产物。 TAF被水解 羧酸酯酶1（CES1），而TDF主要由CES2水解。 TAF和TDF列在 约75％用于两种预防前预防的抗HIV方案 （准备）和治疗作为预防 （TASP）。我们的初步研究还表明，芬太尼增加了趋化因子共同的表达 受体并导致HIV感染增加。该项目的中心假设是基于代谢 Tenofovir药物与酒精+/-芬太尼的相互作用可靠地表明有效性 和替诺福韦方案的安全性，并作为发展依从性监测和的基础 准备/TASP干预策略。具体目的是：（1）表征代谢物特征和 机械生物标志物和（2）确定酒精+/-芬太尼改变代谢的重要性。到 确定特定于酒精+/-芬太尼，人类原发性肝细胞和周围的代谢物特征 血液单核细胞（PBMC） 在存在或没有 酒精（+/-芬太尼）；代谢物特征将由LC-MS/MS确定。改变的代谢产物 体内研究将确定签名。转录组分析将在两个组织上进行（例如， PBMC）和单细胞水平（选定样品）以识别机械生物标志物。建立 意义，肝细胞和PBMC将与具有或不含酒精+/-的替诺福韦型孵育 将监测芬太尼，病毒载荷和细胞毒性。代谢物签名和 具有功效和安全性的生物标志物将在体内确定。最后，来自HIV阴性患者的PBMC 暴露于PREP，PREP+酒精或Prep+酒精/芬太尼时，将针对HIV感染进行测试。这 体外，体内和体内研究设计的结合确保了高科学严格。