Objective Patients with alcohol-associated hepatitis (AH) have a high mortality. Alcohol exacerbates liver damage by inducing gut dysbiosis, bacterial translocation and inflammation, which is characterised by increased numbers of circulating and hepatic neutrophils. Design In this study, we performed tandem mass tag (TMT) proteomics to analyse proteins in the faeces of controls (n=19), patients with alcohol-use disorder (AUD; n=20) and AH (n=80) from a multicentre cohort (InTeam). To identify protein groups that are disproportionately represented, we conducted over-representation analysis using Reactome pathway analysis and Gene Ontology to determine the proteins with the most significant impact. A faecal biomarker and its prognostic effect were validated by ELISA in faecal samples from patients with AH (n=70), who were recruited in a second and independent multicentre cohort (AlcHepNet). Result Faecal proteomic profiles were overall significantly different between controls, patients with AUD and AH (principal component analysis p=0.001, dissimilarity index calculated by the method of Bray-Curtis). Proteins that showed notable differences across all three groups and displayed a progressive increase in accordance with the severity of alcohol-associated liver disease were predominantly those located in neutrophil granules. Over-representation and Reactome analyses confirmed that differentially regulated proteins are part of granules in neutrophils and the neutrophil degranulation pathway. Myeloperoxidase (MPO), the marker protein of neutrophil granules, correlates with disease severity and predicts 60-day mortality. Using an independent validation cohort, we confirmed that faecal MPO levels can predict short-term survival at 60 days. Conclusions We found an increased abundance of faecal proteins linked to neutrophil degranulation in patients with AH, which is predictive of short-term survival and could serve as a prognostic non-invasive marker. Data are available in a public, open access repository. All .RAW data and .MZML data are deposited at massive.ucsd.edu, study ID: MassIVE MSV000094610.

中文翻译：

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粪便蛋白质组学将中性粒细胞脱颗粒与酒精相关性肝炎患者的死亡率联系起来


目的 酒精相关性肝炎 （AH） 患者死亡率高。酒精通过诱发肠道菌群失调、细菌易位和炎症来加剧肝脏损伤，其特征是循环和肝脏中性粒细胞数量增加。设计 在这项研究中，我们进行了串联质量标签 （TMT） 蛋白质组学，以分析来自多中心队列 （InTeam） 的对照 （n=19）、酒精使用障碍患者 （AUD;n=20） 和 AH （n=80） 粪便中的蛋白质。为了识别代表性过高的蛋白质组，我们使用 Reactome 通路分析和基因本体进行了过度代表性分析，以确定影响最显着的蛋白质。通过 ELISA 在 AH 患者 （n=70） 的粪便样本中验证粪便生物标志物及其预后影响，这些患者被招募到第二个独立的多中心队列 （AlcHepNet） 中。结果 对照组、AUD 和 AH 患者粪便蛋白质组学特征总体上存在显著差异 （主成分分析 p=0.001，差异指数通过 Bray-Curtis 方法计算）。在所有三组中表现出显着差异并根据酒精相关肝病的严重程度显示进行性增加的蛋白质主要是位于中性粒细胞颗粒中的蛋白质。过度表达和 Reactome 分析证实，差异调节蛋白是中性粒细胞和中性粒细胞脱颗粒途径中颗粒的一部分。髓过氧化物酶 （MPO） 是中性粒细胞颗粒的标志蛋白，与疾病严重程度相关，可预测 60 天死亡率。使用独立的验证队列，我们证实粪便 MPO 水平可以预测 60 天的短期生存率。 结论 我们发现 AH 患者与中性粒细胞脱颗粒相关的粪便蛋白丰度增加，这可预测短期生存，可作为预后非侵入性标志物。数据在公共、开放访问存储库中可用。都。RAW 数据和 .MZML 数据存放在 massive.ucsd.edu，研究 ID：MassIVE MSV000094610。