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MAD2B promotes tubular epithelial-to-mesenchymal transition and renal tubulointerstitial fibrosis via Skp2

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Abstract

The mitotic arrest deficient protein MAD2B is a well-defined anaphase-promoting complex/cyclosome (APC/C) inhibitor and a small subunit of DNA polymerase zeta. It is critical for mitotic control and DNA repair. However, the pathological role of MAD2B in kidney diseases has not been fully elucidated. In the present study, we aim to explore the role of MAD2B in the pathogenesis of renal tubulointerstitial fibrosis (TIF) and the underlying mechanism. By immunofluorescence and immunohistochemistry, we found an obvious MAD2B enhancement in tubular area of TIF patients and unilateral ureteral obstruction (UUO) mice. In vitro, transforming growth factor-β1 (TGF-β1) induced a time-dependent MAD2B accumulation prior to tubular epithelial-to-mesenchymal transition (EMT) in a rat proximal tubular epithelial cell line, NRK-52E. Knocking down MAD2B using siRNA dramatically inhibited TGF-β1-induced tubular EMT process and subsequent extracellular matrix (ECM) production. We also found that Skp2, a confirmed APC/C-CDH1 substrate and E-cadherin destroyer, was increased in TGF-β1-treated proximal tubular epithelial cells, which could be blocked by MAD2B depletion. In addition, Skp2 expression was also found to be increased in the renal tubular area of UUO mice. Locally knocking down MAD2B expression in the renal cortex using lentiviral transfection inhibited Skp2 expression, tubular EMT, and subsequent ECM accumulation. Taken together, our data suggests a pro-fibrotic role of MAD2B in the pathogenesis of tubular EMT and TIF by inducing Skp2 expression. MAD2B might be a potential target of promising interventions for renal TIF.

Key messages

  • Renal fibrosis activates MAD2B expression in renal tubules of human and mouse.

  • TGF-β1 contributes to MAD2B enhancement in rat tubular epithelial cells.

  • MAD2B depletion alleviates renal tubulointerstitial fibrosis in vivo and in vitro.

  • MAD2B promotes EMT transition in rat tubular epithelial cells by inducing Skp2.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 81170662, No. 31200872, No. 81470964, No. 81570671, No. 81471490, and No. 81522010), a grant from the Wuhan Science and Technology Bureau (No. 2015060101010039), and a Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20130142110064).

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Authors and Affiliations

  1. Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China

    Hui Tang, Di Fan, Chun-Tao Lei, Chen Ye, Pan Gao, Shan Chen, Hua Su & Chun Zhang

  2. Department of Neurobiology, School of Basic Medical Sciences, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Xian-Fang Meng

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  1. Hui Tang
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Correspondence to Chun Zhang.

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The use of parts of mouse kidney biopsies and human renal biopsies for research purposes was approved by the Ethics Committee of Huazhong University of Science and Technology. Animals were treated humanely, and all the procedures were performed according to the guidelines for use and care of laboratory animals of National Institutes of Health (NIH), and approved by the Animal Care and Use Committee (ACUC) of Tongji Medical College.

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Tang, H., Fan, D., Lei, CT. et al. MAD2B promotes tubular epithelial-to-mesenchymal transition and renal tubulointerstitial fibrosis via Skp2. J Mol Med 94, 1297–1307 (2016). https://doi.org/10.1007/s00109-016-1448-6

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  • DOI: https://doi.org/10.1007/s00109-016-1448-6

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