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MafG/MYH9-LCN2 axis promotes liver fibrosis through inhibiting ferroptosis of hepatic stellate cells

Abstract

Hepatic stellate cells (HSCs) secrete extracellular matrix for collagen deposition, contributing to liver fibrosis. Ferroptosis is a novel type of programmed cell death induced by iron overload-dependent lipid peroxidation. Regulation of ferroptosis in hepatic stellate cells (HSCs) may have therapeutic potential for liver fibrosis. Here, we found that Maf bZIP transcription factor G (MafG) was upregulated in human and murine liver fibrosis. Interestingly, MafG knockdown increased HSCs ferroptosis, while MafG overexpression conferred resistance of HSCs to ferroptosis. Mechanistically, MafG physically interacted with non-muscle myosin heavy chain IIa (MYH9) to transcriptionally activate lipocalin 2 (LCN2) expression, a known suppressor for ferroptosis. Site-directed mutations of MARE motif blocked the binding of MafG to LCN2 promoter. Re-expression of LCN2 in MafG knockdown HSCs restored resistance to ferroptosis. In bile duct ligation (BDL)-induced mice model, we found that treatment with erastin alleviated murine liver fibrosis by inducing HSC ferroptosis. HSC-specific knowdown MafG based on adeno-associated virus 6 (AAV-6) improved erastin-induced HSC ferroptosis and alleviation of liver fibrosis. Taken together, MafG inhibited HSCs ferroptosis to promote liver fibrosis through transcriptionally activating LCN2 expression. These results suggest that MafG/MYH9-LCN2 signaling pathway could be a novel targets for the treatment of liver fibrosis.

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Fig. 1: The expression level of MafG is upregulated in liver fibrosis.
Fig. 2: MafG knockdown promotes erastin-mediated HSC ferroptosis.
Fig. 3: MafG overexpression inhibits erastin-induced ferroptosis in HSC.
Fig. 4: LCN2 is an effector gene of MafG in regulating ferroptosis.
Fig. 5: Silencing MafG contributes to ferroptosis of HSC through LCN2.
Fig. 6: MYH9 is required for MafG in transcriptional regulation of LCN2.
Fig. 7: Block of MafG contributes to the IKE-induced HSC ferroptosis in vivo.

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Data availability

All data generated during this study are available within the article and its supplementary information files can be acquired from the corresponding author upon reasonable request. Human MafG ChIP-seq data were obtained from GEO datasets (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE92076). RNA-seq data was acquired from GEO dataset (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE14323).

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Funding

Funding

This work was supported by grants from the National Natural Science Foundation of China (81974074, 82172654 and 82070632), the Natural Science Foundation of Hunan Province (2023JJ30916 and 2023JJ41020), Hunan Provincial Science and Technology Department (2021RC4012 and R2023124).

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Conceptualization, YC and TL; Methodology, YD, LL; Software, DZ and HZ; Validation, YD, LL, YT. and XT; Formal analysis, YF and MG; Investigation, HY and BY; Resources, YC; Data curation, YD and YC; Writing – Original Draft Preparation, YD: Writing – Review & Editing, YC and TL; Visualization, YD; Supervision, YC and TL; Project Administration, YC and TL. All authors have read and approved the final version of the manuscript.

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Correspondence to Ting Liu or Yongheng Chen.

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All experimental procedures for mice were carried out under the Central South University Animal Care and Use Committee criteria. The research protocol for human liver samples conformed to the guidelines of ethical committees of the Xiangya Hospital of Central South University.

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Deng, Y., Lu, L., Zhu, D. et al. MafG/MYH9-LCN2 axis promotes liver fibrosis through inhibiting ferroptosis of hepatic stellate cells. Cell Death Differ (2024). https://doi.org/10.1038/s41418-024-01322-5

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