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tRF3-IleAAT reduced extracellular matrix synthesis in diabetic kidney disease mice by targeting ZNF281 and inhibiting ferroptosis

Acta Pharmacologica Sinica volume 45, pages 1032–1043 (2024)Cite this article

Abstract

It is well established that the synthesis of extracellular matrix (ECM) in mesangial cells is a major determinant of diabetic kidney disease (DKD). Elucidating the major players in ECM synthesis may be helpful to provide promising candidates for protecting against DKD progression. tRF3-IleAAT is a tRNA-derived fragment (tRF) produced by nucleases at tRNA-specific sites, which is differentially expressed in the sera of patients with diabetes mellitus and DKD. In this study we investigated the potential roles of tRFs in DKD. Db/db mice at 12 weeks were adapted as a DKD model. The mice displayed marked renal dysfunction accompanied by significantly reduced expression of tRF3-IleAAT and increased ferroptosis and ECM synthesis in the kidney tissues. The reduced expression of tRF3-IleAAT was also observed in high glucose-treated mouse glomerular mesangial cells. We administered ferrostatin-1 (1 mg/kg, once every two days, i.p.) to the mice from the age of 12 weeks for 8 weeks, and found that inhibition of the onset of ferroptosis significantly improved renal function, attenuated renal fibrosis and reduced collagen deposition. Overexpression of tRF3-IleAAT by a single injection of AAV carrying tRF3-IleAAT via caudal vein significantly inhibited ferroptosis and ECM synthesis in DKD model mice. Furthermore, we found that the expression of zinc finger protein 281 (ZNF281), a downstream target gene of tRF3-IleAAT, was significantly elevated in DKD models but negatively regulated by tRF3-IleAAT. In high glucose-treated mesangial cells, knockdown of ZNF281 exerted an inhibitory effect on ferroptosis and ECM synthesis. We demonstrated the targeted binding of tRF3-IleAAT to the 3’UTR of ZNF281. In conclusion, tRF3-IleAAT inhibits ferroptosis by targeting ZNF281, resulting in the mitigation of ECM synthesis in DKD models, suggesting that tRF3-IleAAT may be an attractive therapeutic target for DKD.

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Fig. 1: tRF3-IleAAT is downregulated in DKD models.
Fig. 2: tRF3-IleAAT overexpression alleviates ECM synthesis in DKD models.
Fig. 3: Inhibition of ferroptosis alleviates ECM synthesis in vivo and in vitro.
Fig. 4: tRF3-IleAAT overexpression alleviates ferroptosis in DKD models.
Fig. 5: Knockdown of the tRF3-IleAAT downstream target gene ZNF281 alleviates ferroptosis in HG-induced mesangial cells.
Fig. 6: tRF3-IleAAT directly targets ZNF281 and negatively regulates its expression.

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Acknowledgements

This study was supported by the National Natural Science Foundation of China (No. 81970664), the Natural Science Foundation of Jiangsu Province (Nos. BK20211385 and BK20191082), the Dedicated Fund for Pediatric Medical Research of Jiangsu Province Medical Association (Nos. SYH-32034-0073 and SYH-32034-0085), and the 789 Outstanding Talent Program of SAHNMU (No. 789ZYRC202090251). We are grateful to the experimental center of the Second Affiliated Hospital of Nanjing Medical University for providing the platform.

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  1. These authors contributed equally: Yun-yang Qiao, Jia-ling Ji

Authors and Affiliations

  1. Department of Pediatric Nephrology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210003, China

    Yun-yang Qiao, Gao-ting Qu, Shan-wen Li, Xing-yue Li, Ran Jin, Yin-fang Li & Hui-min Shi

  2. Department of Pediatrics, the Fourth Affiliated Hospital of Nanjing Medical University, Nanjing, 210031, China

    Yun-yang Qiao, Jia-ling Ji & Ai-qing Zhang

  3. Department of Science and Education, the Affiliated Jiangning Hospital of Nanjing Medical University, Nanjing, 211199, China

    Wei-ling Hou

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Contributions

AQZ and HMS conceived and designed the experiments. YYQ, JLJ and WLH performed the experiments and analysed the data. YYQ wrote the manuscript. JLJ helped edit the manuscript, and WLH drew graphic materials. AQZ and HMS revised the manuscript and confirmed the authenticity of all the raw data. GTQ, SWL, XYL, RJ and YFL contributed to the data analysis and graphical abstract. All authors have read and endorsed the final manuscript.

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Correspondence to Hui-min Shi or Ai-qing Zhang.

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Qiao, Yy., Ji, Jl., Hou, Wl. et al. tRF3-IleAAT reduced extracellular matrix synthesis in diabetic kidney disease mice by targeting ZNF281 and inhibiting ferroptosis. Acta Pharmacol Sin 45, 1032–1043 (2024). https://doi.org/10.1038/s41401-024-01228-5

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