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/艾滋病仍然是一个重大的全球健康问题，慢性肝病已成为导致艾滋病的主要原因 艾滋病毒死亡率。人们认识到几个影响因素，包括抗艾滋病毒药物的肝毒性、 肝炎病毒和广泛饮酒的双重感染。同时使用其他成瘾物质的可能性 物质如 因为芬太尼含量很高。过量饮酒与艾滋病病毒感染增加直接相关 负荷、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， 已知酒精和芬太尼会改变药物代谢酶和转运蛋白的功能，这一点至关重要 抗HIV药物的有效性和安全性的决定因素。事实上，我们已经证明乙醇（酒精） 暴露减少了水解 富马酸替诺福韦艾拉酚胺 (TAF) 并产生了一种新的代谢物： 乙基TAF。该代谢物是一种混合分子，部分来自 TAF，部分来自乙醇。 TAF是一个新的 TDF（富马酸替诺福韦二吡呋酯）的版本，具有降低的器官毒性，并且两种替诺福韦前药都是 水解并随后磷酸化产生具有治疗活性的代谢物。 TAF 被水解 羧酸酯酶-1 (CES1)，而 TDF 主要由 CES2 水解。 TAF 和 TDF 列于 约 75% 的抗 HIV 方案用于暴露前预防 (PrEP) 和治疗作为预防 （塔斯普）。我们的初步研究还表明芬太尼增加了趋化因子辅酶的表达 受体并导致艾滋病毒感染增加。该项目的中心假设是基于新陈代谢的 替诺福韦药物与酒精+/-芬太尼的相互作用产生可靠地表明疗效的特征变化 和替诺福韦治疗方案的安全性，并作为开发依从性监测和治疗的基础 PrEP/TasP 干预策略。具体目标是：(1) 表征代谢特征和 机制生物标志物，(2) 确定酒精+/-芬太尼改变代谢的重要性。到 确定酒精+/-芬太尼、人类原代肝细胞和外周血的特定代谢特征 血液单核细胞 (PBMC) 在存在或不存在的情况下将接受替诺福韦方案治疗 酒精（+/- 芬太尼）；代谢物特征将通过 LC-MS/MS 确定。改变的代谢物 签名将通过体内研究来确定。将在两个组织中进行转录组分析（例如， PBMC）和单细胞水平（选定的样本）来识别机械生物标志物。建立 重要的是，肝细胞和 PBMC 将与含或不含酒精的替诺福韦方案一起孵育 +/- 芬太尼、病毒载量和细胞毒性将受到监测。代谢特征和代谢特征的联系 具有有效性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/芬太尼将进行离体 HIV 感染测试。这 体外、离体和体内研究设计的结合确保了高度的科学严谨性。