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: 10208640
• 负责人: Gyongyi Szabo
• 金额: $ 43.56万
• 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/10208640
• 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; Prognosis; 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; biomarker signature; candidate marker; clinical application; cohort; disease natural history; exosome; extracellular vesicles; insight; large datasets; liver injury; microvesicles; mortality; novel marker; 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.

摘要 酒精性肝炎（AH）死亡率高，由于缺乏可靠的治疗方法， 肝损伤、炎症和感染的标志物，决定临床结果。细胞外囊泡（EV） 与血清相关生物标志物相比， 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 疾病的独特标志。虽然这些不同EV群体的表征是一个新兴领域， EV作为潜在生物标志物的用途在临床应用中迅速发展。我们的初步结果表明 AH中EV的数量增加，这些EV具有不同的RNA，micro-RNA和蛋白质 货物相比，正常对照，表明其潜在的生物标志物的发现。然而， 区分没有肝病的酗酒者和患有酒精性肝炎的人的生物学 是未知的。在这个应用中，我们建议采取两步的方法来研究电动汽车货物作为一个潜在的 酒精性肝炎的生物标志物。UH 2阶段将分两步关注发现阶段：首先，定义 EV分离、RNA和蛋白质样品制备的最佳方法，第二，中试蛋白质组学， 转录组分析将确定酒精性肝炎的潜在生物标志物候选者， 正常对照（DASH样品）。第二阶段，UH 3，将验证蛋白质组学和RNA-seq 目前在来自AlcHepNet的较大AH患者队列中，UH 2期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）。蛋白组 3组充分表征的人类队列的RNA转录组学谱将鉴定EV相关的 AH的特征不同于没有肝脏疾病的HD和正常对照。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的生存率和/或临床结局。来自这些“组学”分析的大型数据集将 建立一个独特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的研究。