Kreimeyer Henriette, Gonzalez Carlos G, Fondevila Marcos F, Hsu Cynthia L, Hartmann Phillipp, Zhang Xinlian, Stärkel Peter, Bosques-Padilla Francisco, Verna Elizabeth C, Abraldes Juan G, Brown Robert S, Vargas Victor, Altamirano Jose, Caballería Juan, Shawcross Debbie L, Louvet Alexandre, Lucey Michael R, Mathurin Philippe, Garcia-Tsao Guadalupe, Bataller Ramón, Investigators AlcHepNet, Gonzalez David J, Schnabl Bernd
Department of Medicine, University of California San Diego, La Jolla, California, USA.
Department of Pharmacology, University of California San Diego, La Jolla, California, USA.
Gut. 2024 Dec 10;74(1):103-115. doi: 10.1136/gutjnl-2024-332730.
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.
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).
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.
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.
酒精性肝炎（AH）患者死亡率很高。酒精通过诱导肠道菌群失调、细菌易位和炎症加剧肝损伤，其特征是循环和肝脏中性粒细胞数量增加。
在本研究中，我们进行了串联质谱标签（TMT）蛋白质组学分析，以分析来自一个多中心队列（InTeam）的对照组（n = 19）、酒精使用障碍（AUD；n = 20）患者和AH患者（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过表达分析，以确定影响最显著的蛋白质。通过酶联免疫吸附测定（ELISA）在来自第二个独立多中心队列（AlcHepNet）的AH患者（n = 70）的粪便样本中验证粪便生物标志物及其预后作用。
对照组、AUD患者和AH患者的粪便蛋白质组图谱总体上有显著差异（主成分分析p = 0.001，采用Bray-Curtis方法计算的差异指数）。在所有三组中显示出显著差异并随着酒精性肝病严重程度逐渐增加的蛋白质主要是位于中性粒细胞颗粒中的蛋白质。过表达和Reactome分析证实，差异调节的蛋白质是中性粒细胞颗粒和中性粒细胞脱颗粒途径的一部分。中性粒细胞颗粒的标记蛋白髓过氧化物酶（MPO）与疾病严重程度相关，并可预测60天死亡率。使用独立验证队列，我们证实粪便MPO水平可预测60天的短期生存率。
我们发现AH患者粪便中与中性粒细胞脱颗粒相关的蛋白质丰度增加，这可预测短期生存率，并可作为一种预后非侵入性标志物。
