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Inhibition of USP10 induces myeloma cell apoptosis by promoting cyclin D3 degradation

Acta Pharmacologica Sinica volume 44, pages 1920–1931 (2023)Cite this article

Abstract

The cell cycle regulator cyclin D3 (CCND3) is highly expressed in multiple myeloma (MM) and it promotes MM cell proliferation. After a certain phase of cell cycle, CCND3 is rapidly degraded, which is essential for the strict control of MM cell cycle progress and proliferation. In the present study, we investigated the molecular mechanisms regulating CCND3 degradation in MM cells. By utilizing affinity purification-coupled tandem mass spectrometry, we identified the deubiquitinase USP10 interacting with CCND3 in human MM OPM2 and KMS11 cell lines. Furthermore, USP10 specifically prevented CCND3 from K48-linked polyubiquitination and proteasomal degradation, therefore enhancing its activity. We demonstrated that the N-terminal domain (aa. 1–205) of USP10 was dispensable for binding to and deubiquitinating CCND3. Although Thr283 was important for CCND3 activity, it was dispensable for CCND3 ubiquitination and stability modulated by USP10. By stabilizing CCND3, USP10 activated the CCND3/CDK4/6 signaling pathway, phosphorylated Rb, and upregulated CDK4, CDK6 and E2F-1 in OPM2 and KMS11 cells. Consistent with these findings, inhibition of USP10 by Spautin-1 resulted in accumulation of CCND3 with K48-linked polyubiquitination and degradation that synergized with Palbociclib, a CDK4/6 inhibitor, to induce MM cell apoptosis. In nude mice bearing myeloma xenografts with OPM2 and KMS11 cells, combined administration of Spautin-l and Palbociclib almost suppressed tumor growth within 30 days. This study thus identifies USP10 as the first deubiquitinase of CCND3 and also finds that targeting the USP10/CCND3/CDK4/6 axis may be a novel modality for the treatment of myeloma.

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Fig. 1: USP10 interacts with CCND3.
Fig. 2: USP10 prevents CCND3 from K48-linked polyubiquitination.
Fig. 3: USP10 stabilizes CCND3 in MM cells.
Fig. 4: The N-terminal aa. 1–205 fragment is dispensable for USP10 to bind to and deubiquitinate CCND3.
Fig. 5: USP10 stabilizes both wild-type and T283A mutant CCND3.
Fig. 6: USP10 activates the CCND3 signaling pathway and promotes MM cell proliferation.
Fig. 7: Inhibition of USP10 synergizes with CDK4/6 inhibitor to induce MM cell apoptosis.

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Acknowledgements

This study was supported in part by National Natural Science Foundation of China (#81970194 and #82170176 to XLM), the National Key Research and Development Program of China (#2022YFC2705003 to XLM), Guangzhou Medical University Discipline Construction Funds (Basic Medicine) (Grant No. JCXKJS2022A05 to XLM), and Department of Education of Guangdong Province of China (Grant No. 2021ZDZX2009 to XLM).

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  1. These authors contributed equally: Yu-jia Xu, Kun Zeng.

Authors and Affiliations

  1. Department of Hematology, the First Affiliated Hospital & GMU-GIBH Joint School of Life Sciences, the Guangdong-Hong Kong-Macao Joint Laboratory for Cell Fate Regulation and Diseases, Guangzhou Medical University, Guangzhou, 510120, China

    Yu-jia Xu, Ying-hui Ye, Xiao-ting Zhu, Zhen-qian Huang & Xin-liang Mao

  2. Guangdong & Guangzhou Key Laboratory of Protein Modification and Degradation, School of Basic Medical Sciences, Guangzhou Medical University, Guangzhou, 511436, China

    Yu-jia Xu, Zi-ying Sun & Xin-liang Mao

  3. Department of Pharmacology, Soochow University, Suzhou, 215123, China

    Kun Zeng, Ying Ren, Bi-yin Cao, Zu-bin Zhang, Guo-qiang Xu & Xin-liang Mao

  4. School of Medicine, Southern University of Science and Technology, Shenzhen, 518055, China

    Chen-yu Mao

  5. Science and Technology Innovation Center, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China

    Xiao-ting Zhu & Xin-liang Mao

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Contributions

XLM designed the study. KZ, YJX, and XLM developed methods. KZ, YJX, YR, ZQH, CYM, YHY, XTZ, and ZYS performed experiments. XLM, ZQH, YJX, and GQX analyzed and interpreted data. XLM and YJX wrote and composed the manuscript. XLM, ZQH, BYC, and ZBZ administrated the projected.

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Correspondence to Zhen-qian Huang or Xin-liang Mao.

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Xu, Yj., Zeng, K., Ren, Y. et al. Inhibition of USP10 induces myeloma cell apoptosis by promoting cyclin D3 degradation. Acta Pharmacol Sin 44, 1920–1931 (2023). https://doi.org/10.1038/s41401-023-01083-w

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