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


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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  1. Zhang Q, Sakamoto K, Wagner KU. D-type Cyclins are important downstream effectors of cytokine signaling that regulate the proliferation of normal and neoplastic mammary epithelial cells. Mol Cell Endocrinol. 2014;382:583–92.

    Article  CAS  PubMed  Google Scholar 

  2. Fabris S, Agnelli L, Mattioli M, Baldini L, Ronchetti D, Morabito F, et al. Characterization of oncogene dysregulation in multiple myeloma by combined FISH and DNA microarray analyses. Genes Chromosomes Cancer. 2005;42:117–27.

    Article  CAS  PubMed  Google Scholar 

  3. Agnelli L, Bicciato S, Mattioli M, Fabris S, Intini D, Verdelli D, et al. Molecular classification of multiple myeloma: a distinct transcriptional profile characterizes patients expressing CCND1 and negative for 14q32 translocations. J Clin Oncol. 2005;23:7296–306.

    Article  CAS  PubMed  Google Scholar 

  4. Kuehl WM, Bergsagel PL. Early genetic events provide the basis for a clinical classification of multiple myeloma. Hematology Am Soc Hematol Educ Program. 2005;1:346–52.

    Article  Google Scholar 

  5. Bergsagel PL, Kuehl WM, Zhan F, Sawyer J, Barlogie B, Shaughnessy J Jr. Cyclin D dysregulation: an early and unifying pathogenic event in multiple myeloma. Blood. 2005;106:296–303.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Chen BB, Glasser JR, Coon TA, Mallampalli RK. F-box protein FBXL2 exerts human lung tumor suppressor-like activity by ubiquitin-mediated degradation of cyclin D3 resulting in cell cycle arrest. Oncogene. 2012;31:2566–79.

    Article  CAS  PubMed  Google Scholar 

  7. Yoshida A, Choi J, Jin HR, Li Y, Bajpai S, Qie S, et al. Fbxl8 suppresses lymphoma growth and hematopoietic transformation through degradation of cyclin D3. Oncogene. 2021;40:292–306.

    Article  CAS  PubMed  Google Scholar 

  8. Russell A, Thompson MA, Hendley J, Trute L, Armes J, Germain D. Cyclin D1 and D3 associate with the SCF complex and are coordinately elevated in breast cancer. Oncogene. 1999;18:1983–91.

    Article  CAS  PubMed  Google Scholar 

  9. Davis MI, Pragani R, Fox JT, Shen M, Parmar K, Gaudiano EF, et al. Small molecule inhibition of the ibiquitin-specific protease USP2 accelerates cyclin D1 degradation and leads to cell cycle arrest in colorectal cancer and mantle cell lymphoma models. J Biol Chem. 2016;291:24628–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Magiera K, Tomala M, Kubica K, De Cesare V, Trost M, Zieba BJ, et al. Lithocholic acid hydroxyamide destabilizes cyclin D1 and induces G0/G1 arrest by inhibiting deubiquitinase USP2a. Cell Chem Biol. 2017;24:458–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sun T, Xu YJ, Jiang SY, Xu Z, Cao BY, Sethi G, et al. Suppression of the USP10/CCND1 axis induces glioblastoma cell apoptosis. Acta Pharmacol Sin. 2021;42:1338–46.

    Article  CAS  PubMed  Google Scholar 

  12. Gennaro VJ, Stanek TJ, Peck AR, Sun Y, Wang F, Qie S, et al. Control of CCND1 ubiquitylation by the catalytic SAGA subunit USP22 is essential for cell cycle progression through G1 in cancer cells. Proc Natl Acad Sci USA. 2018;115:E9298–307.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang W, Huang X, Xin HB, Fu M, Xue A, Wu ZH. TRAF family member-associated NF-κB activator (TANK) inhibits genotoxic nuclear factor κB activation by facilitating deubiquitinase USP10-dependent deubiquitination of TRAF6 ligase. J Biol Chem. 2015;290:13372–85.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Zhang Z, Tong J, Tang X, Juan J, Cao B, Hurren R, et al. The ubiquitin ligase HERC4 mediates c-Maf ubiquitination and delays the growth of multiple myeloma xenografts in nude mice. Blood. 2016;127:1676–86.

    Article  CAS  PubMed  Google Scholar 

  15. Zhuang H, Ren Y, Mao C, Zhong Y, Zhang Z, Cao B, et al. Induction of zinc finger protein RNF6 auto-ubiquitination for the treatment of myeloma and chronic myeloid leukemia. J Biol Chem. 2022;298:102314.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Huang X, Zhang Q, Lou Y, Wang J, Zhao X, Wang L, et al. USP22 deubiquitinates CD274 to suppress anticancer immunity. Cancer Immunol Res. 2019;7:1580–90.

    Article  CAS  PubMed  Google Scholar 

  17. Tiedemann RE, Mao X, Shi CX, Zhu YX, Palmer SE, Sebag M, et al. Identification of kinetin riboside as a repressor of CCND1 and CCND2 with preclinical antimyeloma activity. J Clin Invest. 2008;118:1750–64.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. He Y, Jiang S, Mao C, Zheng H, Cao B, Zhang Z, et al. The deubiquitinase USP10 restores PTEN activity and inhibits non-small cell lung cancer cell proliferation. J Biol Chem. 2021;297:101088.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Zhang M, Hu C, Tong D, Xiang S, Williams K, Bai W, et al. Ubiquitin-specific peptidase 10 (USP10) deubiquitinates and stabilizes MutS homolog 2 (MSH2) to regulate cellular sensitivity to DNA damage. J Biol Chem. 2016;291:10783–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Yuan J, Luo K, Zhang L, Cheville JC, Lou Z. USP10 regulates p53 localization and stability by deubiquitinating p53. Cell. 2010;140:384–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Naderi S, Gutzkow KB, Låhne HU, Lefdal S, Ryves WJ, Harwood AJ, et al. cAMP-induced degradation of cyclin D3 through association with GSK-3beta. J Cell Sci. 2004;117:3769–83.

    Article  CAS  PubMed  Google Scholar 

  22. Låhne HU, Kloster MM, Lefdal S, Blomhoff HK, Naderi S. Degradation of cyclin D3 independent of Thr-283 phosphorylation. Oncogene. 2006;25:2468–76.

    Article  PubMed  Google Scholar 

  23. Liu J, Xia H, Kim M, Xu L, Li Y, Zhang L, et al. Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13. Cell. 2011;147:223–34.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Niesvizky R, Badros AZ, Costa LJ, Ely SA, Singhal SB, Stadtmauer EA, et al. Phase 1/2 study of cyclin-dependent kinase (CDK)4/6 inhibitor palbociclib (PD-0332991) with bortezomib and dexamethasone in relapsed/refractory multiple myeloma. Leuk Lymphoma. 2015;56:3320–8.

    Article  CAS  PubMed  Google Scholar 

  25. Zhan F, Huang Y, Colla S, Stewart JP, Hanamura I, Gupta S, et al. The molecular classification of multiple myeloma. Blood. 2006;108:2020–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Diehl JA, Zindy F, Sherr CJ. Inhibition of cyclin D1 phosphorylation on threonine-286 prevents its rapid degradation via the ubiquitin-proteasome pathway. Genes Dev. 1997;11:957–72.

    Article  CAS  PubMed  Google Scholar 

  27. Qie S, Diehl JA. Cyclin D degradation by E3 ligases in cancer progression and treatment. Semin Cancer Biol. 2020;67:159–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Shan J, Zhao W, Gu W. Suppression of cancer cell growth by promoting cyclin D1 degradation. Mol Cell. 2009;36:469–76.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Deng M, Yang X, Qin B, Liu T, Zhang H, Guo W, et al. Deubiquitination and activation of AMPK by USP10. Mol Cell. 2016;61:614–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Bhattacharya U, Neizer-Ashun F, Mukherjee P, Bhattacharya R. When the chains do not break: The role of USP10 in physiology and pathology. Cell Death Dis. 2020;11:1033.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Bomberger JM, Barnaby RL, Stanton BA. The deubiquitinating enzyme USP10 regulates the post-endocytic sorting of cystic fibrosis transmembrane conductance regulator in airway epithelial cells. J Biol Chem. 2009;284:18778–89.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Takahashi M, Kitaura H, Kakita A, Kakihana T, Katsuragi Y, Nameta M, et al. USP10 is a driver of ubiquitinated protein aggregation and aggresome formation to inhibit apoptosis. iScience. 2018;9:433–50.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Yu M, Fang ZX, Wang WW, Zhang Y, Bu ZL, Liu M, et al. Wu-5, a novel USP10 inhibitor, enhances crenolanib-induced FLT3-ITD-positive AML cell death via inhibiting FLT3 and AMPK pathways. Acta Pharmacol Sin. 2021;42:604–12.

    Article  CAS  PubMed  Google Scholar 

  34. Toogood PL, Harvey PJ, Repine JT, Sheehan DJ, VanderWel SN, Zhou H, et al. Discovery of a potent and selective inhibitor of cyclin-dependent kinase 4/6. J Med Chem. 2005;48:2388–406.

    Article  CAS  PubMed  Google Scholar 

  35. Wang H, Nicolay BN, Chick JM, Gao X, Geng Y, Ren H, et al. The metabolic function of cyclin D3-CDK6 kinase in cancer cell survival. Nature. 2017;546:426–30.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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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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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).

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