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DNA 5mC and RNA m6A modification successively facilitates the initiation and perpetuation stages of HSC activation in liver fibrosis progression

Cell Death & Differentiation volume 30, pages 1211–1220 (2023)Cite this article

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Abstract

Hepatic stellate cells (HSC) are key effector cells in liver fibrosis. Upon stimulation, the quiescent HSC undergoes complex morphological and functional changes to transdifferentiate into activated collagen-producing myofibroblasts. DNA/RNA methylations (5mC/m6A) are both implicated to participate in hepatic fibrosis, yet their respective roles and specific targets in HSC activation remain elusive. Here, we demonstrate that 5mC is indispensable for the initiation stage of HSC activation (myofibroblast transdifferentiation), whereas m6A is essential for the perpetuation stage of HSC activation (excessive ECM production). Mechanistically, DNA 5mC hypermethylation on the promoter of SOCS3 and PPARγ genes leads to STAT3-mediated metabolic reprogramming and lipid loss in the initiation stage. RNA m6A hypermethylation on the transcripts of major collagen genes enhances the mRNA stability in a YTHDF1-dependent manner, which contributes to massive ECM production. Vitamin A-coupled YTHDF1 siRNA alleviates CCl4-induced liver fibrosis in mice through HSC-specific inhibition of collagen production. HIF-1α, which is transactivated by STAT3, serves as a bridge linking the initiation and the perpetuation stages through transactivating YTHDF1. These findings indicate successive roles of DNA 5mC and RNA m6A modification in the progression of HSC activation, which provides new drug targets for epigenetic therapy of liver fibrosis.

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Fig. 1: The time-course analysis characterizes the initiation and perpetuation stages of HSC activation.
Fig. 2: 5mC/m6A modification exhibits stage-specific increase during HSC activation.
Fig. 3: STAT3 is activated in the initiation stage via 5mC-mediated SOCS3 suppression.
Fig. 4: YTHDF1-mediated m6A mechanism contributes to excessive collagen production.
Fig. 5: HIF-1α links the initiation and the perpetuation stages through transactivating YTHDF1.
Fig. 6: Proposed working model of the proposed mechanism in this study.

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Data availability

Data supporting the present study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (32272962, 31972638), and the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. MOE Joint International Research Laboratory of Animal Health and Food Safety, Nanjing Agricultural University, Nanjing, Jiangsu, PR China

    Yue Feng, Shihui Guo, Yulan Zhao, Lei Wu, Yimin Jia & Ruqian Zhao

  2. Key Laboratory of Animal Physiology & Biochemistry, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, Jiangsu, PR China

    Yue Feng, Shihui Guo, Yulan Zhao, Jiayu Qian, Lei Wu, Yimin Jia & Ruqian Zhao

  3. Center for Translational Biomedical Research, UNCG, Kannapolis, NC, USA

    Haibo Dong

  4. College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu, PR China

    Yun Hu

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Contributions

RZ and YF formulated the idea and designed the experiments. YF, SG and YZ performed and analyzed most of the experiments. HD and YH helped with animal work and provided technical help. JQ, LW and YJ performed some experiments and helped with data analysis. RZ and YF wrote the manuscript.

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Correspondence to Ruqian Zhao.

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The authors declare no competing interests.

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The animal experiment was approved by the Animal Ethics Committee of Nanjing Agricultural University. The sampling procedure followed the “Guidelines on Ethical Treatment of Experimental Animals” (2006) No. 398 set by the Ministry of Science and Technology, China.

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Feng, Y., Guo, S., Zhao, Y. et al. DNA 5mC and RNA m6A modification successively facilitates the initiation and perpetuation stages of HSC activation in liver fibrosis progression. Cell Death Differ 30, 1211–1220 (2023). https://doi.org/10.1038/s41418-023-01130-3

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