Steixner-Kumar Agnes Anna, Santacruz Diana, Geiger Tobias, Rust Werner, Böttner Dennis, Krenkel Oliver, Bahrami Ehsan, Okafo George, Barth Thomas F E, Haenle Mark, Kratzer Wolfgang, Schlingeloff Patrycja, Schmidberger Julian, Neubauer Heike, Dick Alec, Werner Markus, Simon Eric
Department of Global Computational Biology and Digital Sciences, Boehringer Ingelheim Pharma GmbH & Co.KG, Biberach, Germany.
Department of Cardiometabolic Research, Boehringer Ingelheim Pharma GmbH & Co.KG, Biberach, Germany.
Hepatol Commun. 2025 Apr 21;9(5). doi: 10.1097/HC9.0000000000000643. eCollection 2025 May 1.
Metabolic dysfunction-associated steatotic liver disease (MASLD) progresses to metabolic dysfunction-associated steatohepatitis (MASH) and is a major cause of liver cirrhosis. Although liver inflammation is the hallmark feature of MASH versus MASLD, the involvement of the peripheral immune cell compartments in disease progression is poorly understood, and single-cell profiles of peripheral immune cells in MASLD/MASH are not known.
Patients with MASLD/MASH and healthy volunteers have been prospectively enrolled in a cross-sectional study. Patients have been histologically stratified and further characterized by liver bulk RNA sequencing (RNA-Seq). Peripheral immune cells from patients and control blood samples have been comprehensively profiled using bulk and single RNA-Seq.
Twenty-two patients with fibrosis stage less than F3 have been histologically stratified into patients with low, medium, and high disease activity scores (NAFLD activity score [NAS]). In contrast to fibrosis, the NAS group correlated with noninvasive imaging readouts and blood biomarkers of liver damage and inflammation (ALT, AST). The prevalence of type 2 diabetes and obesity increased with the NAS stage. Bulk RNA-seq profiling of patient liver biopsies revealed gene signatures that were positively and negatively associated with NAS. Known marker genes for liver fibrosis where upregulated on RNA level. Blood bulk RNA-seq showed only moderate differences in patients versus healthy controls. In contrast, single-cell analysis of white blood cells revealed multiple alterations of immune (sub-)populations, including an increased abundance of immature B cells and myeloid suppressor cells in patients with MASLD/MASH as compared to healthy controls.
The study gives new insights into the pathophysiology of MASLD/MASH already manifesting relatively early in peripheral immune cell compartments. This opens new avenues for the development of new biomarker diagnostics and disease therapies.
代谢功能障碍相关脂肪性肝病（MASLD）会进展为代谢功能障碍相关脂肪性肝炎（MASH），是肝硬化的主要病因。尽管肝脏炎症是MASH区别于MASLD的标志性特征，但外周免疫细胞区室在疾病进展中的作用尚不清楚，MASLD/MASH中外周免疫细胞的单细胞图谱也未知。
MASLD/MASH患者和健康志愿者被前瞻性纳入一项横断面研究。患者已根据组织学进行分层，并通过肝脏整体RNA测序（RNA-Seq）进行进一步表征。使用整体和单细胞RNA-Seq对患者的外周免疫细胞和对照血样进行全面分析。
22例纤维化分期小于F3的患者已根据组织学分为疾病活动评分（非酒精性脂肪性肝病活动评分[NAS]）低、中、高的患者。与纤维化不同，NAS组与肝脏损伤和炎症的非侵入性成像读数及血液生物标志物（ALT、AST）相关。2型糖尿病和肥胖的患病率随NAS分期增加。患者肝脏活检的整体RNA-seq分析揭示了与NAS呈正相关和负相关的基因特征。已知的肝纤维化标志物基因在RNA水平上上调。血液整体RNA-seq显示患者与健康对照之间只有适度差异。相比之下，白细胞的单细胞分析揭示了免疫（亚）群的多种改变，包括与健康对照相比，MASLD/MASH患者中未成熟B细胞和髓系抑制细胞的丰度增加。
该研究为MASLD/MASH的病理生理学提供了新的见解，其在外周免疫细胞区室中相对较早地表现出来。这为开发新的生物标志物诊断方法和疾病治疗开辟了新途径。
