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Updated: Apr 29, 2026
Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation
Published on: April 3, 2017
Single-cell profiling reveals hepatic monocyte-derived macrophages heterogeneity during steatotic liver disease
Haozhe Xu1,2, Lu Yang3, Peiyang Fang1,2
1Medical Research Center, Beijing Institute of Respiratory Medicine, Beijing Chao-Yang Hospital, Capital Medical University, Beijing, China.
Background:
Monocyte-derived macrophages (MoMFs) play key roles in liver inflammation and fibrogenesis, with their heterogeneity affecting metabolic dysfunction-associated steatotic liver disease (MASLD) progression. However, current therapeutic strategies targeting macrophage-mediated inflammation have shown limited clinical efficacy in MASLD.
Methods:
We conducted single-cell RNA sequencing (scRNA-seq) of liver-infiltrating 7AAD-CD45+Ly6G-CD11bhiF4/80int MoMFs from NCD-fed and MCD-fed mice. Monocle 2 and CellChat analyses explored developmental trajectories and intercellular interactions, respectively.
Results:
Seven distinct clusters (c0-c6) with unique molecular signatures were identified. Beyond the classical CD206+ M2-polarized MoMFs, we identified 2 distinct subsets: CCR3+ (c3) MoMFs with enhanced pro-inflammatory and oxidative stress activities, and CCR7+ (c4) MoMFs with specialized antigen-presenting capacity in MASLD mouse livers. Although CCR2+ MoMFs are classically considered pro-inflammatory, our study revealed that the predominant CD14+CCR2+ (c0) MoMFs exhibit additional functional roles in fibrosis, lipid accumulation, and antigen presentation. Our pseudotime and in vitro data demonstrate that resident basal c1 MoMFs are phenotypically plastic, capable of acquiring CD14+/CCR7+ markers and transitioning toward c0-like and c4-like states, implying a potential intrahepatic origin for these subsets in diet-induced steatohepatitis. Notably, these MoMFs subsets and their dynamic changes during disease progression were conserved between the mouse models and human MASLD samples.
Conclusions:
Our study systematically characterized the heterogeneity and dynamic changes in intrahepatic MoMFs during MASLD progression. Resident c1 MoMFs are plastic and could be a local source for CD14+/CCR7+ subsets in MASLD without relying solely on bone marrow recruitment. These findings provide new insights into the therapeutic strategies that target macrophage-mediated inflammation in MASLD.
Insights
Monocyte-derived macrophages (MoMFs) exhibit diverse subtypes in metabolic dysfunction-associated steatotic liver disease (MASLD). Resident MoMFs are plastic and can differentiate into subsets crucial for MASLD progression, offering new therapeutic targets.
Area of Science:
- Immunology
- Hepatology
- Cell Biology
Background:
- Monocyte-derived macrophages (MoMFs) are critical in liver inflammation and fibrogenesis.
- Macrophage heterogeneity influences metabolic dysfunction-associated steatotic liver disease (MASLD) progression.
- Current therapies for MASLD targeting macrophage inflammation show limited success.
Purpose of the Study:
- To systematically characterize the heterogeneity and dynamic changes of intrahepatic MoMFs in MASLD.
- To identify distinct MoMF subsets and their functional roles in MASLD pathogenesis.
- To explore the origin and plasticity of MoMFs in diet-induced liver disease.
Main Methods:
- Single-cell RNA sequencing (scRNA-seq) of liver-infiltrating MoMFs from NCD- and MCD-fed mice.
- Computational analyses (Monocle 2, CellChat) to determine developmental trajectories and intercellular interactions.
- In vitro experiments to validate MoMF plasticity and differentiation.
Main Results:
- Seven distinct MoMF clusters were identified, including classical M2, pro-inflammatory CCR3+ (c3), and antigen-presenting CCR7+ (c4) subsets.
- CD14+CCR2+ (c0) MoMFs, classically pro-inflammatory, also play roles in fibrosis, lipid accumulation, and antigen presentation.
- Resident basal c1 MoMFs are phenotypically plastic, transitioning to c0-like and c4-like states, suggesting an intrahepatic origin.
- Identified MoMF subsets and their dynamics are conserved between mouse models and human MASLD.
Conclusions:
- Intrahepatic MoMF heterogeneity significantly impacts MASLD progression.
- Resident MoMFs are plastic and can serve as a local source for pro-inflammatory and antigen-presenting subsets in MASLD.
- These findings offer novel insights for developing targeted therapies against macrophage-driven inflammation in MASLD.
