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AUF1 protects against ferroptosis to alleviate sepsis-induced acute lung injury by regulating NRF2 and ATF3

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Abstract

Background

The AU-rich element (ARE)-binding factor 1 (AUF1) acts as a switch for septic shock, although its underlying mechanisms remain largely unknown. In this study, we examined the biological significance and potential molecular mechanism of AUF1 in regulating ferroptosis in sepsis-induced acute lung injury (ALI).

Methods

Alveolar epithelial cells (AECs) challenged with ferroptosis-inducing compounds and cecum ligation and puncture (CLP)-induced ALI were used as the in vitro and in vivo model, respectively. The stability of AUF1 and its degradation by ubiquitin–proteasome pathway were examined by cycloheximide chase analysis and co-immunoprecipitation assay. The regulation of AUF1 on nuclear factor E2-related factor 2 (NRF2) and activation transcription factor 3 (ATF3) was explored by RNA immunoprecipitation (RIP), RNA pull-down, and mRNA stability assays. Functionally, the effects of altering AUF1, NRF2 or ATF3 on ferroptosis in AECs or ALI mice were evaluated by measuring cell viability, lipid peroxidation, iron accumulation, and total glutathione level.

Results

AUF1 was down-regulated in AECs challenged with ferroptosis-inducing compounds, both on mRNA and protein levels. The E3 ubiquitin ligase FBXW7 was responsible for protein degradation of AUF1 during ferroptosis. By up-regulating NRF2 and down-regulating ATF3, AUF1 antagonized ferroptosis in AECs in vitro. In the CLP-induced ALI model, the survival rate of AUF1 knockout mice was significantly reduced and the lung injuries were aggravated, which were related to the enhancement of lung ferroptosis.

Conclusions

FBXW7 mediates the ubiquitination and degradation of AUF1 in ferroptosis. AUF1 antagonizes ferroptosis by regulating NRF2 and ATF3 oppositely. Activating AUF1 pathway may be beneficial to the treatment of sepsis-induced ALI.

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All data generated or analyzed during this study are included in this published article.

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

Funding

This work was supported by National Natural Science Foundation of China (No.31872800), Foundation of The Third Affiliated Hospital of Guangzhou Medical University (No.2019Z02) and Foundation of Guangzhou Science and Technology Burea (No.202102010089).

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Author notes

  1. Yichun Wang and Diyu Chen co-first authors.

Authors and Affiliations

  1. Department of Critical Care Medicine, The Third Affiliated Hospital of Guangzhou Medical University, No.63, Duobao Road, Liwan District, Guangdong, 510150, Guangzhou, People’s Republic of China

    Yichun Wang, Mingwang Jia, Fang Peng & Feifei Guo

  2. Department of Obstetrics and Gynecology, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, Guangdong, 510150, Guangzhou, People’s Republic of China

    Diyu Chen

  3. Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangdong, 510150, Guangzhou, People’s Republic of China

    Han Xie & Xiaofang Sun

  4. Department of Surgery, UT Southwestern Medical Center, Dallas, TX, 75390, USA

    Daolin Tang

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Contributions

Guarantor of integrity of the entire study: CD, WYC. Study concepts: WYC, TDL. Study design: WYC, TDL, SXF. Definition of intellectual content: WYC, SXF. Literature research: XH, GFF, PF. Experimental studies: GFF, WYC, CDY. Data acquisition: JMW, GFF. Data analysis: XH, SXF. Statistical analysis: JMW, XH. Manuscript preparation: XH, CDY. Manuscript editing: SXF. Manuscript review: WYC, TDL. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yichun Wang or Daolin Tang.

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All animal protocols were reviewed and approved by the Institutional Animal Care and Use Committee of Guangzhou Medical University (Guangzhou, Guangdong, China).

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18_2022_4248_MOESM1_ESM.tif

Supplementary file 1. Fig. S1. Liproxstatin-1 antagonizes the pro-ferroptotic effects of shNRF2. shNRF2 and control shRNA (shNC) lentiviruses were transduced into AECs for 48 h, before the cells were treated with Lirpoxstatin-1 (inhibitor of ferroptosis, 25 nM) for 12 h. Ferroptosis was assessed by measuring cell viability using MTT assay (A), intracellular iron (B), BODIPY staining (C), and total glutathione (D) levels in indicated cells. *P < 0.05, **P < 0.01 and ***P < 0.001.

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Wang, Y., Chen, D., Xie, H. et al. AUF1 protects against ferroptosis to alleviate sepsis-induced acute lung injury by regulating NRF2 and ATF3. Cell. Mol. Life Sci. 79, 228 (2022). https://doi.org/10.1007/s00018-022-04248-8

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