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NEK9-mediated Wnt signalling repressor TLE3 rewires Docetaxel resistance in cancer cells by inducing pyroptosis

British Journal of Cancer volume 133, pages 1428–1440 (2025)Cite this article

Subjects

Abstract

Background

Docetaxel is the most common chemotherapy regimen for several neoplasms, including advanced OSCC (Oral Squamous Cell Carcinoma). Unfortunately, chemoresistance leads to relapse and adverse disease outcomes.

Methods

We performed CRISPR-based kinome screening to identify potential players of Docetaxel resistance. Immunohistochemistry was performed to examine the expression profile of the target gene across tumour tissues. Global transcriptome analysis was performed to determine the molecular mechanism underlying Docetaxel resistance. NEK9 kinase assay was performed to identify a putative kinase inhibitor.

Results

Upon conducting CRISPR-based kinome screening, Never In Mitosis Gene-A Related Kinase-9 (NEK9) was identified as a major player of Docetaxel resistance in OSCC, prostate, and pancreatic cancer lines. NEK9 expression was found to be upregulated in chemotherapy non-responder OSCC patients as compared to responders. NEK9 ablation restores Docetaxel-induced cell death in chemoresistant cells. Mechanistically, we found that NEK9 deletion upregulates Transducin-like enhancer protein 3 (TLE3), which in turn represses Wnt signalling. Fostamatinib was identified as a potent NEK9 inhibitor that overcomes Docetaxel resistance.

Conclusions

Our study demonstrated that NEK9 plays an important role in Docetaxel resistance. The novel combination of NEK9 inhibitor Fostamatinib and Docetaxel needs further clinical investigation in advanced OSCC.

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Fig. 1: CRISPR-based Kinome screening identified NEK9 as a potential driver of Docetaxel resistance.
Fig. 2: Ablation of NEK9 sensitizes cancer cells to Docetaxel.
Fig. 3: NEK9 kinase activity is the driver of Docetaxel resistance.
Fig. 4: NEK9 kinase safeguards cancer cells from Docetaxel-induced pyroptosis.
Fig. 5: RNA sequencing revealed NEK9 as a modulator Wnt signalling pathway via the TLE3 axis.
Fig. 6: Evaluation of Fostamatinib as a potent NEK9 inhibitor to sensitize cancer cells to Docetaxel-mediated pyroptosis.

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Data availability

RNA sequencing of DU145 (a human prostate cancer cell line) NEK9 genetically knocked out cell line compared to its wildtype parental counterpart has been updated in Array Express, and the assigned ArrayExpress accession is “E-MTAB-14795”. All data are available in the main text or the supplementary materials. The URL to the deposited RNA sequencing dataset is provided below: https://www.ebi.ac.uk/fg/annotare/#list:completed.

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Acknowledgements

SAA is a UGC-SRF, SM is UGC-SRF, and PM is CSIR-SRF. We acknowledge the Institute of Life Sciences, Bhubaneswar, intramural support and DBT BT/ INF/22/SP28293/2018 (for imaging facility).

Funding

This work is funded by Anusandhan National Research Foundation (ANRF: CRG/2022/003427) and Indian Council of Medical Research (ICMR: 2021-10182/GENOMICS/ADHOC-BMS).

Author information

Authors and Affiliations

  1. Institute of Life Sciences, Bhubaneswar, Odisha, India

    Shamima Azma Ansari, Sibasish Mohanty, Pallavi Mohapatra, Sudeshna Datta, Mamuni Swain, Rajeeb K. Swain, Sunil K. Raghav & Rupesh Dash

  2. Regional Centre for Biotechnology, Faridabad, India

    Shamima Azma Ansari, Sibasish Mohanty & Pallavi Mohapatra

  3. Kalinga Institute of Industrial Technology, School of Biotechnology, Bhubaneswar, Odisha, India

    Sudeshna Datta

  4. Department of Oral & Maxillofacial Pathology, SCB Dental College & Hospital, Cuttack, Odisha, India

    Rachna Rath

  5. Department of Surgical Oncology, All India Institute of Medical Sciences, Bhubaneswar, Odisha, India

    Dillip K. Muduly

  6. Department of Radiation Oncology, All India Institute of Medical Sciences, Bhubaneswar, Odisha, India

    Saroj K. D. Majumdar

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  1. Shamima Azma Ansari
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Contributions

Conceptualization: RD, SAA. Methodology: SAA, SM, PM, MS, RKS, SR. Investigation: RD, RR, DM, SKDM. Visualization: RD, SAA, SD. Supervision: RD. Writing-original draft: RD, SAA.

Corresponding author

Correspondence to Rupesh Dash.

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The authors declare no competing interests.

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Ethics approval and consent to participate

The study was approved by the Institute Review Board and Human Ethics Committees (HEC) of the Institute of Life Sciences, Bhubaneswar (120/HEC/23) and All India Institute of Medical Sciences, Bhubaneswar (T/EMF/ Surg.Onco/19/03). All study participants provided written informed consent and agreed to participate in this study. Animal-related experiments were conducted as per the ARRIVE reporting guidelines 2.0 and in accordance with the protocol approved by the Institutional Animal Ethics Committee of Institute of Life Sciences, Bhubaneswar (ILS/IAEC-289-AH/ NOV-22). The Institutional Biosafety Committee (IBSC) approved all related experiments.

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Ansari, S.A., Mohanty, S., Mohapatra, P. et al. NEK9-mediated Wnt signalling repressor TLE3 rewires Docetaxel resistance in cancer cells by inducing pyroptosis. Br J Cancer 133, 1428–1440 (2025). https://doi.org/10.1038/s41416-025-03148-5

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