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Inhibition of USP7 induces p53-independent tumor growth suppression in triple-negative breast cancers by destabilizing FOXM1

Cell Death & Differentiation volume 30, pages 1799–1810 (2023)Cite this article

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Abstract

Although numerous studies indicate that inhibition of USP7 suppresses tumor growth by activating p53, the precise mechanism by which USP7 contributes to tumor growth through the p53-independent manner is not well understood. p53 is frequently mutated in most triple-negative breast cancers (TNBC), characterized as the very aggressive form of breast cancers with limited treatment options and poor patient outcomes. Here, we found that the oncoprotein Forkhead Box M1 (FOXM1) acts as a potential driver for tumor growth in TNBC and, surprisingly, through a proteomic screen, we identified USP7 as a major regulator of FOXM1 in TNBC cells. USP7 interacts with FOXM1 both in vitro and in vivo. USP7 stabilizes FOXM1 through deubiquitination. Conversely, RNAi-mediated USP7 knockdown in TNBC cells, dramatically reduced the levels of FOXM1. Moreover, based upon the proteolysis targeting chimera (PROTAC) technology, we generated PU7-1 (protein degrader for USP7-1), as a USP7 specific degrader. PU7-1 induces rapid USP7 degradation at low nanomolar concentrations in cells but shows no obvious effect on other USP family proteins. Strikingly, the treatment of TNBC cells with PU7-1 significantly abrogates FOXM1 functions and effectively suppresses cell growth in vitro. By using xenograft mouse models, we found that PU7-1 markedly represses tumor growth in vivo. Notably, ectopic overexpression of FOXM1 can reverse the tumor growth suppressive effects induced by PU7-1, underscored the specific effect on FOXM1 induced by USP7 inactivation. Together, our findings indicate that FOXM1 is a major target of USP7 in modulating tumor growth in a p53-independent manner and reveals the USP7 degrader as a potential therapeutic tool for the treatment of triple-negative breast cancers.

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Fig. 1: FOXM1 directly interacts with USP7 deubiquitinase.
Fig. 2: USP7 deubiquitinates and stabilizes FOXM1 protein.
Fig. 3: PU7-1 is a potent and selective PROTAC degrader for USP7 protein.
Fig. 4: PU7-1 treatment leads to the FOXM1 degradation and the suppression of its transcriptional targets.
Fig. 5: PU7-1 mediates FOXM1 degradation, cell proliferation inhibition and tumor suppression in TNBC.
Fig. 6: FOXM1 is required for the PU7-1-mediated growth inhibition and tumor suppression in TNBC.

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Data availability

All uncropped blots and chemistry details were provided in supplementary file. All other data supporting the finding of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Dr. Richard Baer for kind support in this project by providing critical reagents. This work was supported by the National Cancer Institute of the National Institutes of Health under Award R35CA253059, RO1CA258390 and R01CA254970 to WG. We acknowledge the support from the Herbert Irving Comprehensive Cancer Center (HICCC; P30 CA13696) and thank the Molecular Pathology and Proteomics of Shared Resources of HICCC. This work utilized the NMR Spectrometer Systems at Mount Sinai acquired with funding from the NIH’s SIG grants 1S10OD025132 and 1S10OD028504. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

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Authors and Affiliations

  1. Institute for Cancer Genetics, and Herbert Irving Comprehensive Cancer Center, Vagelos College of Physicians & Surgeons, Columbia University, 1130 Nicholas Ave, New York, NY, 10032, USA

    Jingjie Yi, Huan Li, Bo Chu, Ning Kon & Wei Gu

  2. Department of Cell Biology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China

    Bo Chu

  3. Mount Sinai Center for Therapeutics Discovery, Departments of Pharmacological Sciences, Oncological Sciences and Neuroscience, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, NY, 10029, USA

    Xiaoping Hu, Jianping Hu, Yan Xiong, H. Umit Kaniskan & Jian Jin

  4. Department of Pathology and Cell Biology, Vagelos College of Physicians & Surgeons, Columbia University, 1130 Nicholas Ave, New York, NY, 10032, USA

    Wei Gu

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  1. Jingjie Yi
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Contributions

Conception and experimental design: JY and WG. Methodology and data acquisition: JY, HL, and BC. Analysis and interpretation of data: JY, HL, BC, and WG. Manuscript writing: JY and WG.

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Correspondence to Wei Gu.

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Competing interests

JJ is a cofounder and equity shareholder in Cullgen, Inc., a scientific cofounder and scientific advisory board member of Onsero Therapeutics, Inc., and a consultant for Cullgen, Inc., EpiCypher, Inc., Accent Therapeutics, Inc, and Tavotek Biotherapeutics, Inc. The Jin laboratory received research funds from Celgene Corporation, Levo Therapeutics, Inc., Cullgen, Inc. and Cullinan Oncology, Inc. The other authors declare that they have no competing financial interests.

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Yi, J., Li, H., Chu, B. et al. Inhibition of USP7 induces p53-independent tumor growth suppression in triple-negative breast cancers by destabilizing FOXM1. Cell Death Differ 30, 1799–1810 (2023). https://doi.org/10.1038/s41418-023-01180-7

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