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Exosomes derived from hepatitis B virus-infected hepatocytes promote liver fibrosis via miR-222/TFRC axis

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Abstract

Exosomal miRNAs activates hepatic stellate cell (HSC) and promote fibrosis. miR-222 was found to be increased in hepatitis B virus (HBV)-infected hepatocytes, and ferroptosis was reported to ameliorate liver fibrosis (LF). Although miR-222 and ferroptosis have been implicated in LF, the association between miR-222 and ferroptosis and how they coordinate to regulate LF are still not explicit. This study investigates the roles of miR-222 and transferrin receptor (TFRC) in LF. Lipid reactive oxygen species (ROS) level was analyzed by flow cytometry. FerroOrange staining was used to measure intracellular iron level. Luciferase reporter assay was adopted to confirm the binding of miR-222 and TFRC. Real-time quantitative PCR and immunoblots were applied to analyze gene and protein expression. The results showed that supplementation of exosomes derived from HBV-infected LO2 cells remarkably enhanced LX-2 cell activation, evidenced by elevated hydroxyprolin (Hyp) secretion and α-SMA and COL1A2 expression. miR-222 was significantly increased in HBV-Exo. Overexpressing miR-222 upregulated cell viability, secretion of Hpy, and expression of α-SMA and COL1A2, which were all blocked by overexpression of TFRC. Further study showed that TFRC was a target of miR-222, and miR-222 promoted LX-2 cell activation through suppressing TFRC-induced ferroptosis in LX-2 cells. Exosomal miR-222 derived from HBV-infected hepatocytes promoted LF through inhibiting TFRC and TFRC-induced ferroptosis. This study emphasizes the significance of miR-222/TFRC axis in LF and suggests new insights in clinical decision making while treating LF.

Graphical abstract

Exosomes derived from HBV-infected LO2 cells promote LX-2 cell activation and liver fibrosis in mouse

Exosomal miR-222 derived from HBV-infected LO2 cells promotes LX-2 cell activation

TFRC is a target of miR-222 and inhibits LX-2 cell activation induced by miR-222

miR-222 promotes LX-2 cell activation through inhibiting TFRC-induced ferroptosis

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Funding

This study was supported by the National Science and Technology Major Project of China (2017ZX10202203-007-005), National Natural Science Foundation of China (81600479 and 81670548), and Wang Baoen liver fibrosis research fund (2020013).

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  1. Qidi Zhang and Ying Qu contributed equally.

Authors and Affiliations

  1. Department of Gastroenterology, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine, No. 650 New Songjiang Road, Shanghai, 201620, China

    Qidi Zhang, Ying Qu, Fei Li, Binghang Li, Zhenghong Li, Yuwei Dong, Lungen Lu & Xiaobo Cai

  2. Department of Gastroenterology, Ruian People’s Hospital, No. 108 Wansong Road, Ruian, 325200, China

    Qingqing Zhang

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  1. Qidi Zhang
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Contributions

Qidi Zhang and Lungen Lu conceived this study. Ying Qu, Qingqing Zhang, Fei Li, Binghang Li, Zhenghong Li, and Yuwei Dong performed the experiments, collected the data, and performed the data analysis. Xiaobo Cai and Ying Qu wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Lungen Lu or Xiaobo Cai.

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The study had approval from the Ethics Committee of Shanghai General Hospital and was conducted in accordance with the Declaration of Helsinki. The animal study was approved by the IACUC and the Ethics Committee of Shanghai General Hospital.

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Zhang, Q., Qu, Y., Zhang, Q. et al. Exosomes derived from hepatitis B virus-infected hepatocytes promote liver fibrosis via miR-222/TFRC axis. Cell Biol Toxicol 39, 467–481 (2023). https://doi.org/10.1007/s10565-021-09684-z

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