Characteristic gene expression in the liver monocyte-macrophage-DC system is associated with the progression of fibrosis in NASH

Xiaoxiao Wang; Zilong Wang; Baiyi Liu; Rui Jin; Yuyun Song; Ran Fei; Xu Cong; Rui Huang; Xiaohe Li; Jia Yang; Lai Wei; Huiying Rao; Feng Liu

doi:10.3389/fimmu.2023.1098056

Characteristic gene expression in the liver monocyte-macrophage-DC system is associated with the progression of fibrosis in NASH

Front Immunol. 2023 Feb 24:14:1098056. doi: 10.3389/fimmu.2023.1098056. eCollection 2023.

Authors

Xiaoxiao Wang¹, Zilong Wang¹, Baiyi Liu¹, Rui Jin¹, Yuyun Song¹, Ran Fei¹, Xu Cong¹, Rui Huang¹, Xiaohe Li¹, Jia Yang¹, Lai Wei², Huiying Rao¹, Feng Liu¹

Affiliations

¹ Peking University People's Hospital, Peking University Hepatology Institute, Beijing Key Laboratory of Hepatitis C and Immunotherapy for Liver Diseases, Beijing International Cooperation Base for Science and Technology on NAFLD Diagnosis, Beijing, China.
² Beijing Tsinghua Changgung Hospital, Tsinghua University, Beijing, China.

Abstract

Background: The monocyte-macrophage-dendritic cell (DC) (MMD) system exerts crucial functions that may modulate fibrogenesis in nonalcoholic steatohepatitis (NASH). In this study, we explored the cell characteristics, distribution and developmental trajectory of the liver MMD system in NASH mice with fibrosis and clarified characteristic genes of the MMD system involved in liver fibrosis progression in NASH mice and patients.

Methods: Single cells in liver tissue samples from NASH and normal mice were quantified using single-cell RNA sequencing (scRNA-seq) analysis. Differentially expressed genes (DEGs) in the MMD system by pseudotime analysis were validated by tyramide signal amplification (TSA)-immunohistochemical staining (IHC) and analyzed by second harmonic generation (SHG)/two-photon excitation fluorescence (TPEF).

Results: Compared with control mice, there were increased numbers of monocytes, Kupffer cells, and DCs in two NASH mouse models. From the transcriptional profiles of these single cells, we identified 8 monocyte subsets (Mono1-Mono8) with different molecular and functional properties. Furthermore, the pseudotime analysis showed that Mono5 and Mono6 were at the beginning of the trajectory path, whereas Mono2, Mono4, Kupffer cells and DCs were at a terminal state. Genes related to liver collagen production were at the late stage of this trajectory path. DEGs analysis revealed that the genes Fmnl1 and Myh9 in the MMD system were gradually upregulated during the trajectory. By TSA-IHC, the Fmnl1 and Myh9 expression levels were increased and associated with collagen production and fibrosis stage in NASH mice and patients.

Conclusions: Our transcriptome data provide a novel landscape of the MMD system that is involved in advanced NASH disease status. Fmnl1 and Myh9 expression in the MMD system was associated with the progression of NASH fibrosis.

Keywords: Fmnl1; Myh9; fibrosis; monocyte-macrophage-DC (MMD) system; nonalcoholic steatohepatitis (NASH); single cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Fibrosis
Gene Expression
Kupffer Cells / metabolism
Mice
Monocytes / metabolism
Non-alcoholic Fatty Liver Disease* / pathology

Grants and funding

This study was supported by the National Natural Science Foundation of China (NSFC) (81870406 HR, 81870407 LW, 82170584 FL), Peking University Medicine Fund of Fostering Young Scholars’ Scientific & Technological Innovation and the Fundamental Research Funds for the Central Universities (BMU2021PYB010 XW), Chinese foundation for hepatitis prevention and control-TianQing liver disease research fund subject (TQGB20210139 XW) and Qin-Min Project (HR).