Yu Xia, Tian Weihong, Bao Xuanwen, Li Zhiwei, Tu Huilan, Xu Xianbin, Chen Jiyang, Zhang Quan, Chen Jianing, Liu Chuan, Dai Xiaomeng, Xu Xinyi, Weng Haoda, Yu Yue, Zhang Xiuding, Zhou Ruoqi, Zhu Wenfeng, Lan Yan, Cheng Jinlin, Xu Min, Wu Wei, Huang Rui, Yang Yida, Sheng Jifang, Guo Jing, Shi Yu
State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, National Clinical Research Center for Infectious Diseases, National Medical Center for Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Department of lmmunology, School of Medicine, Jiangsu University, Zhenjiang, Jiangsu, China.
Hepatology. 2025 Jun 12. doi: 10.1097/HEP.0000000000001427.
Acute-on-chronic liver failure (ACLF) is driven by systemic inflammation and immune dysregulation. This study aims to characterize the immune landscape in ACLF and identify potential therapeutic targets.
We employed single-cell RNA sequencing, vision spatial sequencing, bulk RNA sequencing, and bioinformatics analysis to profile immune cells in ACLF livers compared to acute liver failure, decompensated cirrhosis, and controls. ACLF livers exhibited a distinct immune signature with increased neutrophils, particularly CCL4 + subsets, which drive unresolved inflammation by recruiting additional neutrophils and inflammatory CD14 + S100A9 + monocytes. Resident KCs were reduced, and monocytes differentiated into TREM2 + macrophages with an M1-like pro-inflammatory phenotype, exacerbating inflammation. Additionally, lymphoid cells showed significant dysfunction, with reduced NK cells and relatively expanded T cells exhibiting diminished cytotoxicity or pro-inflammatory cytokine production. Cell-cell communication analysis identified the ANXA1-FPR1 axis as a key interaction between T cells and myeloid cells, serving as a negative feedback mechanism to dampen inflammation. Plasma and hepatic ANXA1 levels were elevated in ACLF patients, correlating with disease severity. In the preclinical model, the ANXA1 peptide Ac2-26 improved liver function, reduced inflammation, and promoted macrophage polarization from M1 to M2. In vitro, Ac2-26 inhibited CCL4-mediated monocyte chemotaxis and M1 polarization, effects partially blocked by the FPR1 inhibitor Randialic acid B. Mechanically, Ac2-26 activated AMPK and inhibited mTOR signaling.
Our findings provide a comprehensive immune profile in ACLF and highlight ANXA1 as a potential therapeutic target for resolving immune dysregulation and improving outcomes in ACLF.
慢加急性肝衰竭（ACLF）由全身炎症和免疫失调驱动。本研究旨在描绘ACLF中的免疫格局并确定潜在的治疗靶点。
我们采用单细胞RNA测序、视觉空间测序、批量RNA测序和生物信息学分析，对ACLF肝脏中的免疫细胞进行分析，并与急性肝衰竭、失代偿期肝硬化及对照组进行比较。ACLF肝脏呈现出独特的免疫特征，中性粒细胞增加，尤其是CCL4+亚群，其通过招募更多中性粒细胞和炎性CD14+S100A9+单核细胞来驱动炎症持续存在。驻留的枯否细胞减少，单核细胞分化为具有M1样促炎表型的TREM2+巨噬细胞，加剧炎症。此外，淋巴细胞表现出明显功能障碍，自然杀伤细胞减少，相对扩增的T细胞表现出细胞毒性或促炎细胞因子产生减少。细胞间通讯分析确定ANXA1-FPR1轴是T细胞与髓样细胞之间的关键相互作用，作为一种负反馈机制来减轻炎症。ACLF患者血浆和肝脏中ANXA1水平升高，与疾病严重程度相关。在临床前模型中，ANXA1肽Ac2-26改善肝功能、减轻炎症，并促进巨噬细胞从M1向M2极化。在体外，Ac2-26抑制CCL4介导的单核细胞趋化和M1极化，FPR1抑制剂兰地酸B可部分阻断这些作用。机制上，Ac2-26激活AMPK并抑制mTOR信号传导。
我们的研究结果提供了ACLF全面的免疫图谱，并突出了ANXA1作为解决免疫失调和改善ACLF预后的潜在治疗靶点。
