Liquid biopsy for alcoholic hepatitis: diagnosis, prognosis and technology development

项目摘要/摘要 该翻译U01的长期目标是定义病理生理学信息和临床 基于细胞外小泡(EV)鞘脂体货物的相关生物标记物 酒精性肝炎的诊断和预后。啊是酒精引起的最严重的形式 肝病死亡率高，卫生保健利用率高，缺乏有效的治疗方法。医疗保健 专业人员受到缺乏预测和预后生物标记物的限制，这些生物标记物可以对AH患者进行风险分层 并根据生物标记物的概况个性化他们的治疗。目前的建议将肝细胞来源的 EVS上的鞘磷脂载体通过提出EVS上的鞘磷脂对巨噬细胞介导的肝脏炎症的影响 从乙醇受损的肝细胞中招募巨噬细胞进入肝脏，导致肝脏损伤和 发炎。我们的初步数据显示，EV在AH时升高，具有以下特征：1)EV 鞘磷脂在AH患者中升高；ii)EV鞘脂水平与临床参数相关，如 国际标准化比率、胆红素和MELD评分；iii)EV激活促炎巨噬细胞 通过鞘脂信号转导的效应器反应；以及iv)肝细胞来源的EV可以通过一种新的 纳米等离子体增强散射(NPES)分析。这导致了一个中心假设，即循环中的EV 鞘磷脂Cargo是一种与发病相关的生物标志物，可用于AH的诊断和预后。 因此，这项建议的目标是：i)开发针对AH的EV鞘脂体学诊断特征；ii) 证明EV鞘脂组学预测急性肝炎死亡率的能力；iii)将EV鞘脂组学扩展到 治疗药物对急性肝炎患者疗效观察的“机制验证”分析 在我们的关联补充临床试验试点U01(RFA-AA-18-005)中提出；以及iv)开发一种新的 NPEs技术检测肝细胞来源诊断急性肝炎的临床试验 血浆微量样品中的EVS。首先，我们将确定EV鞘脂对急性胰腺炎的诊断性能。 先是单站点培训队列，然后是多站点验证队列。第二，我们将直接测试 EV鞘脂可以预测存活率和对治疗的反应的假说预计将减少 从肝细胞释放EV。第三，我们将开发和验证一种新的临床诊断方法 阿。因此，这种多重PIU01授权申请将产生一种液体活检生物标记物，用于AH的诊断和 预后。

PROJECT SUMMARY/ABSTRACT The long-term objective of this translational U01 is to define pathophysiologically informed and clinically relevant biomarkers based on the sphingolipidomic cargo of extracellular vesicles (EVs) that will serve for the diagnosis and prognosis of subjects with alcoholic hepatitis (AH). AH is the severest form of alcohol-induced liver disease with high mortality, high health care utilization, and the absence of effective therapies. Health care professionals are limited by the lack of predictive and prognostic biomarkers that could risk-stratify AH patients and individualize their therapy based on biomarker profiles. The current proposal links hepatocyte-derived sphinogolipid cargo on EVs to macrophage-mediated liver inflammation by proposing that sphingolipids on EVs from ethanol-damaged hepatocytes recruit macrophages in to the liver, resulting in liver injury and inflammation. Our preliminary data show that EVs are elevated in AH with the following features: i) EV sphingolipids are elevated in AH; ii) EV sphingolipid levels correlate with clinical parameters such as the international normalized ratio, bilirubin, and MELD score; iii) EVs activate proinflammatory macrophage effector responses via sphingolipid signaling; and iv) Hepatocyte-derived EVs can be detected via a novel nanoplasmon enhanced scattering (nPES) assay. This has led to the central hypothesis that circulating EV sphingolipid cargo is a pathogenically relevant biomarker for the diagnosis and prognosis of AH. Therefore, the goals of this proposal are to: i) develop an EV sphingolipidomic diagnostic signature for AH; ii) Demonstrate the ability of EV sphingolipidomics to predict mortality in AH; iii) Extend EV sphingolipidomics to a “proof-of-mechanism” assay for the beneficial effects observed in AH patients treated with therapeutic agents proposed in our linked complementary Clinical Trial Pilot U01 (RFA-AA-18-005); and iv) Develop a novel clinical test for the diagnosis of AH utilizing nPES technology to diagnose AH by detecting hepatocyte-derived EVs in plasma microsamples. First, we will determine the diagnostic performance of EV sphingolipids for AH in a single-site training cohort followed by a multi-site validation cohort. Second, we will directly test the hypothesis that EV sphingolipids can predict survival and response to a therapy that is expected to reduce the release of EVs from hepatocytes. Third, we will develop and validate a novel clinical assay for the diagnosis of AH. Thus, this multi-PI U01 grant application will yield a liquid biopsy biomarker for AH diagnosis and prognosis.