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Thiostrepton alleviates experimental colitis by promoting RORγt ubiquitination and modulating dysbiosis

Cellular & Molecular Immunology volume 20, pages 1352–1366 (2023)Cite this article

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Abstract

Thiostrepton (TST) is a natural antibiotic with pleiotropic properties. This study aimed to elucidate the therapeutic effect of TST on experimental colitis and identify its targets. The effect of TST on colon inflammation was evaluated in a dextran sulfate sodium (DSS)-induced colitis model and a T-cell transfer colitis model. The therapeutic targets of TST were investigated by cytokine profiling, immunophenotyping and biochemical approaches. The effect of TST on the gut microbiota and its contribution to colitis were evaluated in mice with DSS-induced colitis that were subjected to gut microbiota depletion and fecal microbiota transplantation (FMT). Alterations in the gut microbiota caused by TST were determined by 16S rDNA and metagenomic sequencing. Here, we showed that TST treatment significantly ameliorated colitis in the DSS-induced and T-cell transfer models. Specifically, TST targeted the retinoic acid-related orphan nuclear receptor RORγt to reduce the production of IL-17A by γδ T cells, type 3 innate lymphoid cells (ILC3s) and Th17 cells in mice with DSS-induced colitis. Similarly, TST selectively prevented the development of Th17 cells in the T-cell transfer colitis model and the differentiation of naïve CD4+ T cells into Th17 cells in vitro. Mechanistically, TST induced the ubiquitination and degradation of RORγt by promoting the binding of Itch to RORγt. Moreover, TST also reversed dysbiosis to control colonic inflammation. Taken together, these results from our study describe the previously unexplored role of TST in alleviating colonic inflammation by reducing IL-17A production and modulating dysbiosis, suggesting that TST is a promising candidate drug for the treatment of IBD.

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Data availability

Raw 16 S rRNA sequencing and metagenomic sequencing data have been deposited in the European Nucleotide Archive (http://www.ebi.ac.uk/ena) with Study No. PRJEB53485. The other data are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFA0507900), the National Natural Science Foundation of China (81802460) and the Natural Science Foundation of Chongqing (CSTB2022NSCQ-MSX0184). We thank Hongwei Li (Third Military Medical University, Chongqing, China) for technical assistance in molecular docking. We thank Dr. Lilin Ye and Dr. Jihang Zhang (Third Military Medical University, Chongqing, China) for the fruitful discussions and review of our manuscript and for helping with the FACS experiment.

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  1. These authors contributed equally: Ya Luo, Cheng Liu, Yuan Luo.

Authors and Affiliations

  1. Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, China

    Ya Luo, Cheng Liu, Jing Li, Changjiang Hu & Shiming Yang

  2. Department of Gastroenterology, The Second Affiliated Hospital of Zunyi Medical University, Zunyi Medical University, Zunyi, 563006, China

    Ya Luo

  3. Department of Immunology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Yuan Luo

  4. Department of Gastroenterology, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, China

    Xianglian Zhang

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YL, CL and YL were involved in the study design and drafted the paper; YL, CL and YL performed all the experiments with the help of XLZ and JL. SMY and CJH devised, coordinated, and supervised the project.

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Correspondence to Changjiang Hu or Shiming Yang.

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Luo, Y., Liu, C., Luo, Y. et al. Thiostrepton alleviates experimental colitis by promoting RORγt ubiquitination and modulating dysbiosis. Cell Mol Immunol 20, 1352–1366 (2023). https://doi.org/10.1038/s41423-023-01085-y

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