Extracellular Vesicles in Alcoholic Liver Disease: Basic and Pre-Clinical Discovery

酒精性肝病中的细胞外囊泡：基础和临床前发现

• 批准号: 10167062
• 负责人: Gyongyi Szabo
• 金额: $ 43.75万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10167062
• 关键词: Alcoholic Hepatitis; Alcoholic Liver Diseases; Alcoholic beverage heavy drinker; Alcohols; Biogenesis; Biological Markers; Biological Process; Biology; Caliber; Cells; Characteristics; Chronic; Clinical; Code; Data; Diagnosis; Disease; Disease Outcome; Future; Hepatitis B; Hepatocyte; Hepatology; Human; Immune; Infection; Inflammation; Literature; Liver diseases; Mass Spectrum Analysis; Mediator of activation protein; MicroRNAs; Observational Study; Organ; Outcome; Patients; Phase; Pilot Projects; Population; Preparation; Proteins; Proteomics; Protocols documentation; Quality Control; RNA; RNA purification; Research Personnel; Research Project Grants; Resources; Sampling; Serum; Source; Translational Research; Untranslated RNA; Whole Blood; alcohol exposure; alcohol research; base; biomarker discovery; candidate marker; clinical application; cohort; disease natural history; exosome; extracellular vesicles; insight; large datasets; liver injury; microvesicles; mortality; novel marker; outcome forecast; potential biomarker; pre-clinical; predict clinical outcome; problem drinker; protein biomarkers; protein purification; tool; transcriptome; transcriptome sequencing; transcriptomics

ABSTRACT Alcoholic hepatitis (AH) has high mortality and management of these patients is limited by the lack of reliable markers of liver injury, inflammation and infection that determine clinical outcomes. Extracellular vesicles (EVs) are novel biomarkers that are more sensitive compared to serum associated-biomarkers because of the stability of biomolecules within the EVs. EVs contain characteristic RNA and protein cargo that can provide a unique signature of disease. While characterization of these different EV populations is an emerging field, the use of EVs as potential biomarkers is rapidly evolving in clinical applications. Our preliminary results suggest that there is an increased number of EVs in AH and these EVs have distinct RNA, micro-RNA and protein cargos compared to normal controls that indicate their potential for biomarker discovery. However, the features of biology that distinguish heavy drinkers with no liver disease from those who present with alcoholic hepatitis are unknown. In this application, we propose to take a two-step approach to study the EV cargo as a potential biomarker in alcoholic hepatitis. The UH2 phase will focus on the discovery phase in two steps: first, to define optimal approaches to EV isolation, RNA and protein sample preparation and, second, pilot proteomics and transcriptome analyses that will identify potential biomarker candidates in alcoholic hepatitis compared to normal controls (DASH samples). The second, UH3, phase will validate the proteomic and RNA-seq characteristic of EVs from the UH2 phase now in a larger cohort of AH patients from the AlcHepNet Observational study and compare those to heavy drinkers (HD) without liver disease (discovery). Proteomic and RNA transcriptomic profiles of the 3 sets of well-characterized human cohorts will identify EV-associated signatures of AH that is different from HD without liver disease and normal controls. Our study will further discover correlations between unique signatures in the protein cargo and/or RNA transcriptome profile of EVs that indicate survival and/or clinical outcomes in AH. The large dataset from these “omics” analyses will establish a unique AH-proteomic-transcriptome interface (AH-PTI) and this will provide invaluable resources for the AlcHepNet investigators and the entire alcohol research field for future hypothesis-driven studies.

抽象的 酒精性肝炎 (AH) 死亡率高，这些患者的治疗因缺乏可靠的治疗方法而受到限制 决定临床结果的肝损伤、炎症和感染标志物。细胞外囊泡 (EV) 是比血清相关生物标志物更敏感的新型生物标志物，因为 EV 内生物分子的稳定性。 EV 含有特征性 RNA 和蛋白质货物，可以提供 疾病的独特特征。虽然这些不同电动汽车群体的表征是一个新兴领域， 使用 EV 作为潜在生物标志物在临床应用中正在迅速发展。我们的初步结果表明 AH 中的 EV 数量增加，并且这些 EV 具有独特的 RNA、micro-RNA 和蛋白质 货物与正常对照相比表明其发现生物标志物的潜力。然而，特点 区分没有肝病的酗酒者和患有酒精性肝炎的酗酒者的生物学原理 未知。在此应用中，我们建议采取两步方法来研究电动汽车货物作为潜在的 酒精性肝炎的生物标志物。 UH2阶段将重点关注发现阶段，分两步：首先，定义 EV 分离、RNA 和蛋白质样品制备以及试点蛋白质组学和蛋白质组学的最佳方法 转录组分析将识别酒精性肝炎的潜在生物标志物候选物 正常对照（DASH 样本）。第二阶段 UH3 将验证蛋白质组学和 RNA 测序 现在来自 AlcHepNet 的更大的 AH 患者队列中 UH2 阶段 EV 的特征 观察性研究并将其与无肝病的重度饮酒者 (HD) 进行比较（发现）。蛋白质组学 3 组特征明确的人类群体的 RNA 转录组图谱将鉴定与 EV 相关的病毒 AH 的特征与无肝病的 HD 和正常对照不同。我们的研究将进一步 发现 EV 的蛋白质货物和/或 RNA 转录组谱中的独特特征之间的相关性 表明 AH 中的生存和/或临床结果。这些“组学”分析的大型数据集将 建立独特的 AH-蛋白质组-转录组接口（AH-PTI），这将为 AlcHepNet 研究人员和整个酒精研究领域的未来假设驱动研究。