PAD4+ neutrophils promote hepatic stellate cell activation and accelerate MASH fibrosis progression viaNET-DNA/TAOK1/MAPK pathways
Jiajia Shen, Shanshan Huang, Yaohui Wang, Qingyuan Wang, Shibo Lin, Wei Guan, Yingyun Gong, Yiming Si, Ming Zhao, Hongwen Zhou, Hui Liang
Jiajia Shen, Shanshan Huang, Yaohui Wang, Qingyuan Wang, Shibo Lin, Wei Guan, Yingyun Gong, Yiming Si, Ming Zhao, Hongwen Zhou, Hui Liang
View: Text | PDF
Research Article Cell biology Hepatology Metabolism

PAD4+ neutrophils promote hepatic stellate cell activation and accelerate MASH fibrosis progression viaNET-DNA/TAOK1/MAPK pathways

Abstract

Neutrophils play a pivotal role in the progression of metabolic dysfunction–associated steatohepatitis (MASH) by mediating inflammatory responses. However, the heterogeneity of neutrophil subsets in MASH and their specific contributions to disease progression remain unclear. In this study, analysis of liver biopsies from 265 patients revealed a strong association between elevated neutrophil counts and MASH severity, particularly fibrosis. Five distinct neutrophil subsets were identified in human liver tissue, with PAD4+ neutrophils serving as key drivers in MASH progression. Mechanistically, PAD4+ neutrophils generate neutrophil extracellular traps (NETs) and activate hepatic stellate cells via the TAOK1-dependent MAPK signaling pathway. Inhibition of PAD4+ neutrophils in vivo attenuated the progression of liver fibrosis without exacerbating liver injury. Collectively, these findings elucidate the pivotal involvement of PAD4+ neutrophils in MASH progression and identify them as promising therapeutic targets for mitigating fibrosis and inflammation.

Authors

Jiajia Shen, Shanshan Huang, Yaohui Wang, Qingyuan Wang, Shibo Lin, Wei Guan, Yingyun Gong, Yiming Si, Ming Zhao, Hongwen Zhou, Hui Liang

×

Figure 4

scRNA-Seq reveals neutrophil heterogeneity in human MASLD liver, with PAD4+ neutrophils correlating with MASH development.

Options: View larger image (or click on image) Download as PowerPoint
scRNA-Seq reveals neutrophil heterogeneity in human MASLD liver, with PA...
(A) Uniform manifold approximation and projection (UMAP) analysis of transcriptional profiles of myeloid cells (n = 15,479) from human MASL (n = 6) and MASH (n = 6) livers, with each cluster represented by a different color. (B) Bar plots showing the proportion of 6 myeloid cell subpopulations in human MASL and MASH livers. (C) UMAP analysis of transcriptional profiles of neutrophils (n = 2,428) from human MASL (n = 6) and MASH (n = 6) livers, with each cluster represented by a different color. (D) Violin plots showing expression levels and distribution of representative marker genes across neutrophil clusters. (E) UMAP analysis showing the distribution of 5 neutrophil subclusters in human MASL and MASH livers. (F) Pseudotime trajectory analysis of neutrophils using Palantir software (https://palantir.readthedocs.io), with cells colored by pseudotime progression. (G) UMAP showing NET formation signature scores in neutrophils, with color intensity reflecting scaled signature scores. Seurat (v4.3) AddModuleScore function was used to calculate NET gene signature scores. (H) UMAP of neutrophils colored by PAD4 cluster in MASL and MASH livers. (I) Flow cytometry analysis of PAD4+ neutrophils from livers of WD/CCl4–fed mice. (J) The proportion of PAD4+ neutrophils within the liver-infiltrating neutrophils in HFD- and WD/CCl4–fed mice. Statistical analyses were performed using 2-tailed Student’s t test. Data are shown as the mean ± SEM.