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LncRNA H19-EZH2 interaction promotes liver fibrosis via reprogramming H3K27me3 profiles

Acta Pharmacologica Sinica volume 44, pages 2479–2491 (2023)Cite this article

Abstract

Liver fibrosis is a wound-healing process characterized by excess formation of extracellular matrix (ECM) from activated hepatic stellate cells (HSCs). Previous studies show that both EZH2, an epigenetic regulator that catalyzes lysine 27 trimethylation on histone 3 (H3K27me3), and long non-coding RNA H19 are highly correlated with fibrogenesis. In the current study, we investigated the underlying mechanisms. Various models of liver fibrosis including Mdr2−/−, bile duct ligation (BDL) and CCl4 mice were adapted. We found that EZH2 was markedly upregulated and correlated with H19 and fibrotic markers expression in these models. Administration of EZH2 inhibitor 3-DZNeP caused significant protective effects in these models. Furthermore, treatment with 3-DZNeP or GSK126 significantly inhibited primary HSC activation and proliferation in TGF-β-treated HSCs and H19-overexpreesing LX2 cells in vivo. Using RNA-pull down assay combined with RNA immunoprecipitation, we demonstrated that H19 could directly bind to EZH2. Integrated analysis of RNA-sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) further revealed that H19 regulated the reprogramming of EZH2-mediated H3K27me3 profiles, which epigenetically promoted several pathways favoring HSCs activation and proliferation, including epithelial-mesenchymal transition and Wnt/β-catenin signaling. In conclusion, highly expressed H19 in chronic liver diseases promotes fibrogenesis by reprogramming EZH2-mediated epigenetic regulation of HSCs activation. Targeting the H19-EZH2 interaction may serve as a novel therapeutic approach for liver fibrosis.

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Fig. 1: Hepatic EZH2 levels are correlated to the severity of liver fibrosis in various mouse models.
Fig. 2: EZH2 inhibitors 3-DZNeP and GSK126 significantly abrogate TGF-β-mediated fibrotic response in HSCs.
Fig. 3: The inhibition of EZH2 significantly alleviates hepatic fibrosis in vivo.
Fig. 4: H19 overexpression promotes EZH2-mediated fibrotic gene expression.
Fig. 5: EZH2 inhibitors 3-DZNeP and GSK126 significantly inhibit H19-induced HSC activation.
Fig. 6: H19 can directly interact with EZH2.
Fig. 7: H19 overexpression epigenetically regulates cell proliferation, apoptosis and EMT pathways.
Fig. 8: H19 overexpression epigenetically enhances WNT signaling and promotes liver fibrosis.

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Acknowledgements

This work was supported by grants from Beijing Nova Program of Science & Technology (Z201100006820025 and Z211100002121167 to RPL); the National High-Level Talents Special Support Program to XJYL; National Natural Science Foundation of China (Grant No. 82274186 to XJYL, and 82274201 to RPL); Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine (Grant No. ZYYCXTD-C-202006 to XJYL).

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  1. These authors contributed equally: Xiao-jiao-yang Li, Fei Zhou

Authors and Affiliations

  1. School of Life Sciences, Beijing University of Chinese Medicine, Beijing, 100029, China

    Xiao-jiao-yang Li, Fei Zhou, Ya-jing Li, Xiao-yong Xue, Jiao-rong Qu, Jia Liu & Jian-zhi Wu

  2. School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing, 100029, China

    Gui-fang Fan, Qi Zheng & Run-ping Liu

  3. The Second Hospital of Shandong University, Ji-nan, 250033, China

    Rong Sun

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Contributions

XJYL and FZ contributed equally to this work. XJYL and RPL conceived the original idea and supervised the study. FZ, XJYL and RPL prepared the manuscript and figures. YJL, XYX, JRQ, JL, GFF, RS, JZW, and QZ conducted all the experiments and performed data analysis. All authors have approved the final manuscript.

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Correspondence to Xiao-jiao-yang Li or Run-ping Liu.

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Li, Xjy., Zhou, F., Li, Yj. et al. LncRNA H19-EZH2 interaction promotes liver fibrosis via reprogramming H3K27me3 profiles. Acta Pharmacol Sin 44, 2479–2491 (2023). https://doi.org/10.1038/s41401-023-01145-z

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