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

项目总结/摘要 本翻译U01的长期目标是定义病理生理学信息和临床 基于细胞外囊泡（EV）的鞘脂组学货物的相关生物标志物，其将用于 酒精性肝炎（AH）患者的诊断和预后。AH是酒精诱导的最严重的形式 肝脏疾病具有高死亡率，高卫生保健利用率和缺乏有效治疗。保健 由于缺乏可对AH患者进行风险分层的预测性和预后性生物标志物， 并根据生物标志物的特征对他们进行个体化治疗。目前的建议将肝细胞来源的 通过提出EV上鞘脂，EV上的鞘脂货物对巨噬细胞介导的肝脏炎症的影响 从乙醇损伤的肝细胞募集巨噬细胞进入肝脏，导致肝损伤， 炎症我们的初步数据显示，EV在AH中升高，具有以下特征：i）EV AH中鞘脂升高; ii）EV鞘脂水平与临床参数相关，例如 国际标准化比率、胆红素和MELD评分; iii）EV激活促炎性巨噬细胞 通过鞘脂信号传导的效应子应答;和iv）肝细胞衍生的EV可以通过新的免疫荧光检测。 纳米等离子体增强散射（nPES）测定。这导致了一个中心假设，即循环EV 鞘脂货物是AH诊断和预后的病理相关生物标志物。 因此，该提案的目标是：i）开发AH的EV鞘脂组学诊断特征; ii） 证明EV鞘脂组学预测AH死亡率的能力; iii）将EV鞘脂组学扩展到 在接受治疗剂治疗的AH患者中观察到的有益作用的"机制证明"试验 在我们的相关补充临床试验试点U01（RFA-AA-18 - 005）中提出;和iv）开发一种新的 利用nPES技术通过检测肝细胞源性 血浆微量样本中的EV。首先，我们将确定EV鞘脂对AH的诊断性能， 单中心培训队列，随后是多中心验证队列。第二，我们将直接测试 假设EV鞘脂可以预测生存率和对预期减少 从肝细胞释放EV。第三，我们将开发和验证一种新的临床试验，用于诊断 啊因此，该多PI U01资助申请将产生用于AH诊断的液体活检生物标志物， 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.