Noninvasive detection of nonalcoholic steatohepatitis (NASH), the progressive form of nonalcoholic fatty liver disease, promises to improve patient screening, accelerate drug trials, and reduce health care costs. On the basis of protease dysregulation of the biological pathways of fibrotic NASH, we developed the Glympse Bio Test System (GBTS) for multiplexed quantification of liver protease activity. GBTS-NASH comprises a mixture of 19 mass-barcoded PEGylated peptides that is administered intravenously and senses liver protease activity by releasing mass-barcoded reporters into urine for analysis by mass spectrometry. To identify a protease signature of NASH, transcriptomic analysis of 355 human liver biopsies identified a 13-protease panel that discriminated clinically relevant NASH ≥F2 fibrosis from F0-F1 with high classification accuracy across two independent patient datasets. We screened 159 candidate substrates to identify a panel of 19 peptides that exhibited high activity for our 13-protease panel. In the choline-deficient, L-amino acid-defined, high-fat diet (CDAHFD) mouse model, binary classifiers trained on urine samples discriminated fibrotic NASH from simple steatosis and healthy controls across a range of nondisease conditions and indicated disease regression upon diet change [area under receiver operating characteristics (AUROCs) > 0.97]. Using a hepatoprotective triple combination treatment (FXR agonist, ACC and ASK1 inhibitors) in a rat model of NASH, urinary classification distinguished F0-F1 from ≥F2 animals and indicated therapeutic response as early as 1 week on treatment (AUROCs >0.91). Our results support GBTS-NASH to diagnose fibrotic NASH via an infusion of peptides, monitor changes in disease severity, and indicate early treatment response.

中文翻译：

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通过基于质量条形码的活动传感器基于肽的尿液监测纤维化非酒精性脂肪性肝炎

非酒精性脂肪性肝炎 (NASH) 的无创检测是非酒精性脂肪性肝病的进行性形式，有望改善患者筛查、加速药物试验并降低医疗保健成本。基于纤维化 NASH 的生物学途径的蛋白酶失调，我们开发了 Glympse 生物测试系统 (GBTS) 用于肝蛋白酶活性的多重量化。GBT-NASH 包含 19 种质量条码 PEG 化肽的混合物，静脉内给药并通过将质量条码报告分子释放到尿液中进行质谱分析来检测肝蛋白酶活性。为了识别 NASH 的蛋白酶特征，对 355 例人类肝脏活检的转录组学分析确定了一个 13 种蛋白酶组，该组可在两个独立的患者数据集中区分临床相关的 NASH ≥F2 纤维化和 F0-F1，具有很高的分类准确度。我们筛选了 159 种候选底物，以确定一组 19 种肽，它们对我们的 13 种蛋白酶组表现出高活性。在胆碱缺乏、L-氨基酸定义的高脂肪饮食 (CDAHFD) 小鼠模型中，对尿液样本进行训练的二元分类器将纤维化 NASH 与简单脂肪变性和健康对照区分开来，并表明饮食会导致疾病消退改变 [接收器操作特性下的区域 (AUROC) > 0.97]。在 NASH 大鼠模型中使用保肝三联疗法（FXR 激动剂、ACC 和 ASK1 抑制剂），尿液分类将 F0-F1 与≥F2 动物区分开来，并在治疗后 1 周表明治疗反应（AUROCs >0.91）。我们的结果支持 GBT-NASH 通过输注肽诊断纤维化 NASH，监测疾病严重程度的变化，并指示早期治疗反应。