A transposon screen identifies enhancement of NF-κB pathway as a mechanism of resistance to eribulin

Abstract

Background

Eribulin mesylate (eribulin) is an efficient microtubule inhibitor that is used for metastatic breast cancer. However, breast cancer can develop resistance to eribulin. This resistance mechanism needs to be elucidated.

Methods

A transposon mutagenesis screen was conducted using a pPB-SB-CMV-puro-SD plasmid and pCMV-PBase transposase. Viability and cytotoxicity were analyzed by MTT assay and flow cytometry, respectively. Real-time PCR and western blot were used for gene expression analysis. In addition, vivo study was also designed to analyze therapy efficiency.

Results

TAB2, which is part of the nuclear factor-kappa B (NF-κB) pathway, was identified as a candidate eribulin-resistant gene. TAB2 down-regulation resulted in significantly lower cell viability and higher cytotoxicity of cells treated with eribulin, while TAB2 up-regulation showed opposite results. Similarly, combination of NF-κB inhibitors [Bay-117082 and QNZ (quinazoline derivative)] with eribulin showed significantly lower cell viability and higher drug cytotoxicity than single agent treatment with eribulin in MDA-MB-231 cells. However, QNZ increased NF-κB activity in MCF7 cells by up-regulating TAB2, which reduced the sensitivity to eribulin. Furthermore, combination of Bay-117082 with eribulin induced greater regression of MDA-MB-231 tumors compared to eribulin monotherapy in vivo.

Conclusions

These results consistently illustrated that TAB2-NF-κB pathway may increases resistance to eribulin in breast cancer models. Moreover, these results support the use of a combination strategy of eribulin with NF-κB inhibitors, and provide evidence that transposon mutagenesis screens are capable of identifying drug-resistant genes.

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Acknowledgements

We thank Dr. Li in kindly providing the transposon plasmid and Kazuhiro Miyao (laboratory assistant) at Keio University for assistance in the experiments.

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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by XT, TH, and TM. The first draft of the manuscript was written by XT and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Tetsu Hayashida.

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Conflict of interest

T. Hayashida and Y. Kitagawa received research grants and lecture fees from Eisai Co., Ltd. A. Nagayama’s immediate family member has a leadership position with Chugai Co., Ltd. and owns stock options of Chugai Co., Ltd. All remaining authors have no conflicts of interest to declare.

Research involving human participants and/or animals

This article does not contain any studies with human participants performed by any of the authors. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of Keio university.

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Teng, X., Hayashida, T., Murata, T. et al. A transposon screen identifies enhancement of NF-κB pathway as a mechanism of resistance to eribulin. Breast Cancer (2021). https://doi.org/10.1007/s12282-021-01224-1

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Keywords

  • Breast cancer
  • Drug resistance
  • Transposon mutagenesis screen
  • Eribulin
  • NF-κB