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Deubiquitylation of Rab35 by USP32 promotes the transmission of imatinib resistance by enhancing exosome secretion in gastrointestinal stromal tumours

Oncogene volume 42, pages 894–910 (2023)Cite this article

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Abstract

Imatinib is a tyrosine kinase inhibitor that is widely used to combat gastrointestinal stromal tumours (GISTs). However, secondary resistance to imatinib is an important challenge in GIST treatment. Recent studies have demonstrated that cancer-derived nanosized exosomes play a key role in intercellular communication, but little is known about the roles of exosomes in imatinib-resistant GISTs. Here, we reveal that exosomes released from imatinib-resistant GISTs transmit drug resistance to imatinib-sensitive tumours. By using iTRAQ technology, we demonstrate that Ras-related protein Rab-35 (Rab35) is upregulated differentially in imatinib-resistant GISTs. Loss of Rab35 decreases exosome secretion, thereby hampering the transmission of imatinib resistance to sensitive tumours. Mechanistically, we showed that the ubiquitin‒proteasome system is involved in elevated Rab35 expression and that ubiquitin-specific protease 32 (USP32), a deubiquitylating enzyme, is bound to Rab35. Further experiments demonstrate that this protease protects Rab35 from proteasomal degradation by reducing Lys48 (K48)-ubiquitination. Additionally, we found that the transcription factor ETV1, which is a lineage survival factor in GISTs, promotes USP32 expression. Collectively, our results reveal that exosomes transmit imatinib resistance in GISTs and that deubiquitylation plays a key role in regulating the transmission process. The USP32-Rab35 axis provides a potential target for interventions to reduce the occurrence of imatinib resistance in GISTs.

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Fig. 1: Exosomes transmit imatinib resistance in GIST cells.
Fig. 2: Rab35 is differentially upregulated in imatinib-resistant GISTs and affects the transmission of resistance by regulating exosome secretion in GIST.
Fig. 3: Ubiquitination modification affects Rab35 protein levels.
Fig. 4: USP32 stabilizes Rab35 at the protein level.
Fig. 5: USP32 deubiquitinates Rab35.
Fig. 6: USP32 regulates exosome secretion in GIST cells.
Fig. 7: Targeting USP32 decreases the transmission of imatinib resistance in vivo.
Fig. 8: ETV1 upregulates USP32 transcription.

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All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Additional data related to this paper may be requested from the authors.

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Acknowledgements

This work was supported by grants from National Natural Science Foundation of China (grant number: 82072708); Youth Program of National Natural Science Foundation of China (grant number: 81902461); Natural Science Foundation of Province (grant number: BK20191495, BK20191073); The Priority Academic Program Development of Jiangsu Higher Education Institutions (grant number: PAPD, JX10231801); Jiangsu Key Medical Discipline (General Surgery) (grant number: ZDXKA2016005).

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  1. These authors contributed equally: Chao Li, Zhishuang Gao, Zhiwei Cui.

Authors and Affiliations

  1. Department of General Surgery, the First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China

    Chao Li, Zhishuang Gao, Zhiwei Cui, Zonghang Liu, Haoyu Sun, Nuofan Wang, Zhongyuan He, Bowen Li, Fengyuan Li, Zheng Li, Linjun Wang, Diancai Zhang, Li Yang, Zekuan Xu & Hao Xu

  2. Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Jiangsu Collaborative Innovation Center for Cancer Personalized Medical University, Nanjing, 211166, China

    Chao Li, Zhishuang Gao, Zhiwei Cui, Zonghang Liu, Haoyu Sun, Nuofan Wang, Zhongyuan He, Bowen Li, Fengyuan Li, Zheng Li, Linjun Wang, Diancai Zhang, Li Yang, Zekuan Xu & Hao Xu

  3. Department of Oncology, the Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China

    Yibo Bian

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Contributions

Conception and design: CL and ZSG; data acquisition, analysis, and interpretation: ZSG, ZWC, and ZHL; investigation: YBB, HYS, NFW, ZYH, ZL, BWL and CL; acquisition of patient specimens: FYL, DCZ, and LY; article drafting and revising: CL and HX; article writing: CL; support of experimental and clinical techniques: LJW and ZKX. All authors approved the final version of the manuscript.

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Correspondence to Zekuan Xu or Hao Xu.

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The collection of specimens and animal handling for the study have been reviewed and approved by the Ethics Committee of the First Affiliated Hospital of Nanjing Medical University.

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Li, C., Gao, Z., Cui, Z. et al. Deubiquitylation of Rab35 by USP32 promotes the transmission of imatinib resistance by enhancing exosome secretion in gastrointestinal stromal tumours. Oncogene 42, 894–910 (2023). https://doi.org/10.1038/s41388-023-02600-1

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