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Molecular Diagnostics

Exosomal transfer of miR-181b-5p confers senescence-mediated doxorubicin resistance via modulating BCLAF1 in breast cancer

British Journal of Cancer volume 128, pages 665–677 (2023)Cite this article

Subjects

Abstract

Background

Doxorubicin resistance represents a major clinical challenge for treating patients with advanced breast cancer (BC). Exosomes, exchanging genetic cargo between heterogeneous populations of tumour cells, have been proposed to mediate drug resistance and cancer progression in other cancer types. However, their specific role in mediating doxorubicin resistance in BC remains unclear. Here, we demonstrate the important role of exosomal miR-181b-5p (exo-miR-181b-5p) in mediating doxorubicin resistance.

Methods

Small-RNA sequencing and bioinformatic analyses were used to screen miRNAs mediating doxorubicin resistance in BC, which were further verified by RT-qPCR. SA-β-gal staining assays allowed us to measure cellular senescence. Exosomes from patients’ serum before and after neoadjuvant chemotherapy were isolated for exo-miR-181b-5p quantification.

Results

Doxorubicin-resistant BC cell lines exhibited upregulated exosomal miR-181b-5p. Addition of exo-miR-181b-5p actively fused with recipient cells and transferred a drug-resistant phenotype. Overexpression of miR-181b-5p downregulated p53/p21 levels and inhibited doxorubicin-induced G1 arrest and senescence by suppressing BCLAF1 expression in vitro. Further, in vivo experiments showed treatment of exo-miR-181b-5p inhibitors exhibited superior tumour control and reversed the doxorubicin-resistance phenotype, accompanied with increased tumoral BCLAF1.

Conclusion

Our data suggests exo-miR-181b-5p as a prognostic biomarker and a therapeutic potential for exo-miR-181b-5p inhibitors in the treatment of doxorubicin-resistant BC patients.

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Fig. 1: Conditioned medium from DOX-resistant cells transfers a senescence-mediated DOX-resistant phenotype in breast cancer cells.
Fig. 2: Exosomes derived from DOX-resistant cells transfer a DOX-resistant phenotype consistent with inhibition of DOX-induced cellular senescence.
Fig. 3: Identification of miR-181b-5p secreted in exosomes by DOX-resistant cells as a key mediator of DOX-resistance transfer in breast cancer cell lines; and correlates with DOX-resistance in patient samples.
Fig. 4: Exosomal miR-181b-5p induces DOX-resistance in recipient cells via downregulation of BCLAF1 and subsequent evasion of senescence mediated by p53/p21.
Fig. 5: Systemically injected exosomes pre-loaded with inhibitors of miR-181b-5p sensitises the response to DOX in vivo.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by The Tianjin Science Foundation (No. 19YFZCSY00030 to JZ); The China Scholarship Council Award (No. 202006940028 to TP, No. 201806010012 to JD); Tianjin Municipal Education Research Project (2020KJ136 to SZ); Tianjin Key Medical Discipline (Specialty) Construction Project (TJYXZDXK-009A); and CRUK Early Detection and Diagnosis Committee Project (No. C1519/A27375 to JMV).

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Author notes

  1. These authors contributed equally: Shaorong Zhao, Teng Pan, Jinhai Deng, Lixia Cao.

Authors and Affiliations

  1. The 3rd Department of Breast Cancer, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Breast Cancer Prevention and Therapy, Tianjin Medical University, Ministry of Education, Tianjin, China

    Shaorong Zhao, Teng Pan, Jingjing Liu, Guanglin Zhou & Jin Zhang

  2. Richard Dimbleby Laboratory of Cancer Research, School of Cancer & Pharmaceutical Sciences, King’s College London, London, UK

    Jinhai Deng, Jose M. Vicencio & Tony Ng

  3. Tianjin Key Laboratory of Tumor Microenvironment and Neurovascular Regulation, Medical College of Nankai University, 300071, Tianjin, China

    Lixia Cao

  4. Cancer Institute, Paul O’Gorman Building, University College London, London, UK

    Jose M. Vicencio & Tony Ng

  5. Cancer Research UK City of London Centre, London, England

    Tony Ng

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Contributions

SZ, TP, JD and LC have contributed equally to this work. JZ is the corresponding author to this work. Conceptualisation, resources, supervision and project administration were done by JZ; Methodology, software, formal analysis, investigation, data curation and visualisation were done by TP, SZ, JD, LC and GZ. Writing—original draft and writing—review & editing were done by JZ, TP, SZ, JD and LC. JMV, JL and TN reviewed and edited the manuscript. The authors reviewed and approved the final manuscript.

Corresponding author

Correspondence to Jin Zhang.

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The authors declare no competing interests.

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The study was conformed to the principles of the Declaration of Helsinki, and authorised by the Medical Ethics Committee of Tianjin Medical University Cancer Institute and Hospital and Tianjin Medical University Cancer Institute and Hospital Laboratory Animal Care and Use Committee.

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Table S1. Breast cancer patient characteristics

Table S2. Sequences used in cell transfection

Supplementary figure legends

Figure S1. Conditioned medium (CM) from doxorubicin-resistant cells suppressed senescence in breast cancer cells.

Figure S2: Uptake of exosomes derived from DOX-resistant cells decreased DOX-induced senescence in breast cancer cells.

41416_2022_2077_MOESM7_ESM.tif

Figure S3. Exosomal miR-181b-5p from DOX-resistant cells is responsible for drug resistance and suppresses cellular senescence.

Figure S4. Exosomal miR-181b-5p induced senescence reduction in recipient cells via downregulation of BCLAF1.

Figure S5. Exosomal miR-181b-5p is related to DOX-resistance in vivo.

Table S3-Cal51vsCALDOX.Differential_analysis_results

Table S4-MCF-7vsMCF-7A02.Differential_analysis_results

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Zhao, S., Pan, T., Deng, J. et al. Exosomal transfer of miR-181b-5p confers senescence-mediated doxorubicin resistance via modulating BCLAF1 in breast cancer. Br J Cancer 128, 665–677 (2023). https://doi.org/10.1038/s41416-022-02077-x

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