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: 10542286
• 负责人: JASON T BLACKARD
• 金额: $ 72.17万
• 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/10542286
• 关键词: AIDS prevention; AIDS/HIV problem; 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; Ethanol; Exposure to; Fentanyl; Foundations; Free Radicals; Fumarates; General Population; Goals; HIV; HIV Infections; HIV Seronegativity; HIV risk; Health; Hepatitis Viruses; Hepatocyte; Hepatotoxicity; Human; Hybrids; Hydrolysis; In Vitro; Incubated; Individual; Injections; 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; alcohol exposure; alcohol use disorder; base; 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.

摘要 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 艾滋病毒--死亡率。人们认识到几个促成因素，包括抗艾滋病毒药物的肝脏毒性， 肝炎病毒的混合感染和广泛的饮酒。同时使用其他成瘾剂的可能性 物质，如 因为芬太尼很高。过量饮酒与HIV病毒的增加有直接联系 参与艾滋病毒危险行为，以及在艾滋病毒预防干预措施中依从性差。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，对 抗艾滋病毒药物有效性和安全性的决定因素。事实上，我们已经证明了乙醇(酒精) 暴露降低了水解酶的活性 替诺福韦，富马酸阿拉芬胺(TAF)，并产生一种新的代谢物： 乙基TAF。该代谢物是一种杂化分子，部分来自TAF，部分来自乙醇。Taf是一种新的 TDF(富马酸替诺福韦)的版本具有较低的器官毒性，并且两种替诺福韦前体药物 水解，然后磷酸化，产生具有治疗活性的代谢物。Taf的水解物是 羧酸酯酶-1(CES1)，而TDF主要由CES2水解酶。中列出了TAF和TDF ~75%的抗HIV方案用于暴露前预防 (准备)和作为预防的治疗 (TasP)。我们的初步研究还表明，芬太尼增加了趋化因子联合- 并导致艾滋病毒感染增加。这个项目的中心假设是，基于新陈代谢 替诺福韦药物与酒精+/-芬太尼的相互作用产生可靠指示疗效的标志性变化 和安全性，并作为制定依从性监测和 PREP/TasP干预策略。具体目标是：(1)表征代谢物特征和 机械性生物标志物，以及(2)确定酒精+/-芬太尼改变代谢的意义。至 确定酒精+/-芬太尼、人原代肝细胞和外周血特异性代谢物特征 血液单个核细胞(PBMC) 将在存在或不存在的情况下使用替诺福韦方案进行治疗 酒精(+/-芬太尼)；代谢物特征将通过LC-MS/MS确定。 签名将通过体内研究来确定。将在两个组织上执行转录组分析(例如， PBMC)和单细胞水平(选定的样本)来识别机械性生物标记物。要建立 重要的是，肝细胞和PBMC将与替诺福韦+或不含酒精+/-方案孵育 芬太尼，以及病毒载量和细胞毒性将被监测。代谢物特征和代谢产物的联系 具有有效性和安全性的生物标志物将在体内被确定。最后，来自HIV阴性患者的PBMCs 暴露于PrEP、PrEP+酒精或PrEP+酒精/芬太尼将在体外测试抗HIV感染。这个 体外、体外和体内研究设计相结合，确保了高度的科学严谨性。