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USP19 potentiates autophagic cell death via inhibiting mTOR pathway through deubiquitinating NEK9 in pancreatic cancer

Cell Death & Differentiation volume 32, pages 702–713 (2025)Cite this article

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Abstract

The ubiquitin-specific protease (USP) family is the largest and most diverse deubiquitinase (DUBs) family and plays a significant role in maintaining cell homeostasis. Dysregulation of USPs has been associated with carcinogenesis of various tumors. We identified that USP19 was downregulated in pancreatic tumor tissues and forced expression of USP19 diminished tumorigenicity of pancreatic cancer. Mechanistically, USP19 directly interacts with and stabilized NEK9 via inhibiting K48-specific polyubiquitination process on NEK9 protein at K525 site through its USP ___domain. Moreover, NEK9 phosphorylates the regulatory associated protein of mTOR (Raptor) at Ser792 and links USP19 to the inhibition of mTORC1 signaling pathway, which further leads to autophagic cell death of pancreatic cancer cells. Inhibition of autophagy by Atg5 knockdown or lysosome inhibitor bafilomycin A1 abolished the decreased malignant phenotype of USP19- and NEK9-overexpressing cancer cells. Importantly, USP19 expression exhibits a positive correlation with NEK9 expression in clinical samples, and low USP19 or NEK9 expression is associated with a worse prognosis. This study revealed that USP19-mediated NEK9 deubiquitylation is a regulatory mechanism for mTORC1 inhibition and provides a therapeutic target for diseases involving mTORC1 dysregulation.

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Fig. 1: USP19 is downregulated in pancreatic cancer and related with a worse prognosis.
Fig. 2: Inhibitory effects of USP19 on proliferation and metastasis of pancreatic cancer cells.
Fig. 3: USP19 binds to and inhibits NEK9 degradation.
Fig. 4: Effects of USP19 on ubiquitination of NEK9.
Fig. 5: USP19 inhibits pancreatic cancer proliferation and metastasis through interacting with and stabling NEK9.
Fig. 6: USP19/NEK9 cascade inhibited Warburg effect and activated autophagy pathway.
Fig. 7: NEK9 binds to and promotes Raptor phosphorylation at Ser792.

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Data availability

All datasets are available from the corresponding author on reasonable request.

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Funding

This work was supported by the grant from the Natural Science Foundation of Jiangsu Province (BK20221416), Clinical Science and Technology Climbing program - “Spark” basic research project (ZJ202124), Young Scholars Fostering Fund of the First Affiliated Hospital of Nanjing Medical University (PY202404) and Innovation Capability Development Project of Jiangsu Province (No. BM2015004). All samples used in current study were obtained from Pancreas Biobank, The First Affiliated Hospital with Nanjing Medical University, which is a part of Jiangsu Biobank of Clinical Resource.

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Author notes

  1. These authors contributed equally: Guangfu Wang, Shangnan Dai, Jin Chen, Kai Zhang.

Authors and Affiliations

  1. Pancreas Center, First Affiliated Hospital of Nanjing Medical University, Nanjing, China

    Guangfu Wang, Shangnan Dai, Kai Zhang, Jinfan Zhang, Fuye Lin, Yong Gao, Lingdi Yin, Kuirong Jiang & Zipeng Lu

  2. Pancreas Institute, Nanjing Medical University, Nanjing, China

    Guangfu Wang, Shangnan Dai, Kai Zhang, Jinfan Zhang, Kunxin Xie, Fuye Lin, Yong Gao, Lingdi Yin, Kuirong Jiang, Yi Miao & Zipeng Lu

  3. Department of Gynecological Oncology, Jiangsu Cancer Hospital, Nanjing, China

    Jin Chen

  4. Department of Biomedical Engineering, University of California, Irvine, CA, USA

    Chenyu Huang

  5. Department of Medicine, University of California, Irvine, CA, USA

    Chenyu Huang

  6. Pancreas Center, The Affiliated BenQ Hospital of Nanjing Medical University, Nanjing, China

    Huijuan Wang & Yi Miao

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Contributions

Conception and design: GFW; data acquisition, analysis, and interpretation: GFW, SND, JC, KZ; investigation: GFW, SND, JC, KZ, JFZ, FYL, HJW; methodology: KZ, ZPL, CYH, LDY; acquisition of patient specimens: JFZ, KXX, FYL, HJW, YG; supervision: ZPL, YM, KRJ; resources: ZPL, YM, JC; and article drafting and revising: GFW, SND. All authors approved the final version of the manuscript.

Corresponding authors

Correspondence to Kuirong Jiang, Yi Miao or Zipeng Lu.

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The authors declare no competing interests.

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All participants provided informed consent. All human tissue research in this study had the approval of ethics committees of the First Affiliated Hospital of Nanjing Medical University (Nanjing, China). All animal experiments were performed in accordance with a protocol approved by the Institutional Animal Care and Use Committee of the Nanjing Medical University.

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Wang, G., Dai, S., Chen, J. et al. USP19 potentiates autophagic cell death via inhibiting mTOR pathway through deubiquitinating NEK9 in pancreatic cancer. Cell Death Differ 32, 702–713 (2025). https://doi.org/10.1038/s41418-024-01426-y

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