Abstract
Chronic rhinosinusitis with nasal polyp (CRSwNP) is a refractory inflammatory disease with epithelial-mesenchymal transition (EMT) as one of the key features. Since ubiquitin modification has been shown to regulate the EMT process in other diseases, targeting ubiquitin ligases may be a potential strategy for the treatment of CRSwNP. In this study we investigated whether certain E3 ubiquitin ligases could regulate the EMT process in CRSwNP, and whether these regulations could be the potential drug targets as well as the underlying mechanisms. After screening the potential drug target by bioinformatic analyses, the expression levels of three potential E3 ubiquitin ligases were compared among the control, eosinophilic nasal polyp (ENP) and non-eosinophilic nasal polyp (NENP) group in clinical samples, and the significant decrement of the expression level of NEDD4L was found. Then, IP-MS, bioinformatics and immunohistochemistry studies suggested that low NEDD4L expression may be associated with the EMT process. In human nasal epithelial cells (hNECs) and human nasal epithelial cell line RPMI 2650, knockdown of NEDD4L promoted EMT, while upregulating NEDD4L reversed this effect, suggesting that NEDD4L inhibited EMT in nasal epithelial cells. IP-MS and Co-IP studies revealed that NEDD4L mediated the degradation of DDR1. We demonstrated that NEDD4L inhibited the β-catenin/HIF-1α positive feedback loop either directly (degrading β-catenin and HIF-1α) or indirectly (mediating DDR1 degradation). These results were confirmed in a murine NP model in vivo. This study for the first time reveals the regulatory role of ubiquitin in the EMT process of nasal epithelial cells, and identifies a novel drug target NEDD4L, which has promising efficacy against both ENP and NENP by suppressing β-catenin/HIF-1α positive feedback loop.
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Acknowledgements
This study was supported by the National Natural Science Foundation of China (No. 82071017, and No. 82271134).
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SYC conceived and designed the study, conducted the experiments and analyzed the data, prepared the figures and drafted the paper. PQL, DXQ, HL, and HQZ provided crucial assistance during the animal experiments. YX edited and revised the manuscript.
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This work was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of Renmin Hospital, Wuhan University (No. WDRY2021-K084). The detailed aims of the study and the planned procedures were explained to all patients, who then signed the informed consent documentation. The Institutional Animal Care and Use Committee of the Renmin Hospital of Wuhan University approved our animal experiments which were conducted in accordance with the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health (License No. WDRM 20211005).
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Chen, Sy., Liu, Pq., Qin, Dx. et al. E3 ubiquitin ligase NEDD4L inhibits epithelial-mesenchymal transition by suppressing the β-catenin/HIF-1α positive feedback loop in chronic rhinosinusitis with nasal polyps. Acta Pharmacol Sin 45, 831–843 (2024). https://doi.org/10.1038/s41401-023-01190-8
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DOI: https://doi.org/10.1038/s41401-023-01190-8