Abstract
Chronic kidney disease (CKD) is a major global health concern and the third leading cause of premature death. Renal fibrosis is the primary process driving the progression of CKD, but the mechanisms behind it are not fully understood, making treatment options limited. Here, we find that the E3 ligase TRIM65 is a positive regulator of renal fibrosis. Deletion of TRIM65 results in a reduction of pathological lesions and renal fibrosis in mouse models of kidney fibrosis induced by unilateral ureteral obstruction (UUO)- and folic acid. Through screening with a yeast-hybrid system, we identify a new interactor of TRIM65, the mammalian cleavage factor I subunit CFIm25 (NUDT21), which plays a crucial role in fibrosis through alternative polyadenylation (APA). TRIM65 interacts with NUDT21 to induce K48-linked polyubiquitination of lysine 56 and proteasomal degradation, leading to the inhibition of TGF-β1-mediated SMAD and ERK1/2 signaling pathways. The degradation of NUDT21 subsequently altered the length and sequence content of the 3′UTR (3′UTR-APA) of several pro-fibrotic genes including Col1a1, Fn-1, Tgfbr1, Wnt5a, and Fzd2. Furthermore, reducing NUDT21 expression via hydrodynamic renal pelvis injection of adeno-associated virus 9 (AAV9) exacerbated UUO-induced renal fibrosis in the normal mouse kidneys and blocked the protective effect of TRIM65 deletion. These findings suggest that TRIM65 promotes renal fibrosis by regulating NUDT21-mediated APA and highlight TRIM65 as a potential target for reducing renal fibrosis in CKD patients.
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Data availability
The data analyzed during this study are included in this published article. Additional supporting data are available from the corresponding authors upon reasonable request.
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Acknowledgements
We appreciate Dr. Hongbin Shu and Dr. Shu Li (School of Medicine, Wuhan University) for providing HA-Ub mutant plasmids. We thank Alan Jiang (The First Affiliated Hospital, Jiangxi Medical College, Nanchang University) for microscopy assistance.
Funding
This work was supported by grants from the National Natural Science Foundation of China (82160133, 31960147, and 32170793), Jiangxi Provincial Natural Science Foundation (20212ACB216005, 20224ACB216013, 20224BAB206007, and 20212BAB206086).
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YL conceived and designed the study, prepared the figures, analyzed data, and participated in the paper writing. SW and XH performed the experiments, analyzed the data, and wrote the first draft of the manuscript. QZhu, TC, JT, TP, and XK conducted experiments during the study. LZ, TX, and QZ bred and genotyped the mice. YZ critically revised the manuscript for important intellectual content. EL reviewed the manuscript. All authors approved and contributed to the manuscript.
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All animal procedures were approved by The Institutional Animal Care and Use Committee of The First Affiliated Hospital of Nanchang University and performed in accordance with the ARRIVE guideline. All methods were carried out in accordance with relevant guidelines and regulations.
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Wei, S., Huang, X., Zhu, Q. et al. TRIM65 deficiency alleviates renal fibrosis through NUDT21-mediated alternative polyadenylation. Cell Death Differ 31, 1422–1438 (2024). https://doi.org/10.1038/s41418-024-01336-z
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DOI: https://doi.org/10.1038/s41418-024-01336-z
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