Translational Therapeutics

Inhibition of interferon-signalling halts cancer-associated fibroblast-dependent protection of breast cancer cells from chemotherapy

Abstract

Background

Triple negative breast cancers (TNBC) have poor prognoses despite aggressive treatment with cytotoxic chemotherapy. Cancer-associated fibroblasts (CAFs) are prominent in tumour stroma. Our hypothesis was that CAFs modulate chemotherapy sensitivity.

Methods

TNBC cells and breast fibroblasts were cultured; survival after chemotherapeutics was assessed using luciferase or clonogenic assays. Signalling was investigated using transcriptomics, reporters, recombinant proteins and blocking antibodies. Clinical relevance was investigated using immunohistochemistry.

Results

Breast CAFs dose-dependently protected TNBC cell lines MDA-MB-231 and MDA-MB-157, but not MDA-MB-468s, from chemotherapy. CAF-induced protection was associated with interferon (IFN) activation. CAFs were induced to express IFNβ1 by chemotherapy and TNBC co-culture, leading to paracrine activation in cancer cells. Recombinant IFNs were sufficient to protect MDA-MB-231 and MDA-MB-157 but not MDA-MB-468 cells. In TNBC patients, IFNβ1 expression in CAFs correlated with cancer cell expression of MX1, a marker of activated IFN signalling. High expression of IFNβ1 (CAFs) or MX1 (tumour cells) correlated with reduced survival after chemotherapy, especially in claudin-low tumours (which MDA-MB-231 and MDA-MB-157 cells represent). Antibodies that block IFN receptors reduced CAF-dependent chemoprotection.

Conclusions

CAF-induced activation of IFN signalling in claudin-low TNBCs results in chemoresistance. Inhibition of this pathway represents a novel method to improve breast cancer outcomes.

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Fig. 1: Breast CAFs, but not NFs, protect some TNBC lines from chemotherapy.
Fig. 2: CAFs stimulate IFN signalling in some co-cultured breast epithelial cell lines.
Fig. 3: Epithelial:fibroblast crosstalk induces IFNβ1 expression in CAFs and IFN signalling in epithelial cells.
Fig. 4: Recombinant IFNs are sufficient to stimulate chemoresistance in MDA-MB-231 and MDA-MB-157 cells.
Fig. 5: In primary cancers, IFNβ1 in CAFs and MX1 in cancer cells correlate with each other and with poor survival.
Fig. 6: Blocking antibodies inhibit CAF-induced chemoprotection.

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Acknowledgements

Fiona Errington-Mais and Andrew Macdonald (both University of Leeds, UK) kindly provided materials for the study.

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Authors

Contributions

R.V.B.—planned project, designed and performed experiments, analysed data. S.J.J.—provided resources, designed and performed experiments, analysed data. M.C.T.—performed experiments. L.M.W.—provided resources, analysed data. A.M.H.—supervised project, performed and oversaw pathology analyses. J.L.T.—supervised and managed project. T.A.H.—supervised and managed project, designed experiments, analysed data, lead manuscript writing. All authors contributed to writing the manuscript.

Corresponding author

Correspondence to Thomas A. Hughes.

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Ethical permissions for use of patient material and data from patients was granted by Leeds (East) REC (references 09/H1326/108 and 06/Q1206/180). Patients recruited and informed consent was taken in line with these permissions. The study was performed in accordance with the Declaration of Helsinki.

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not applicable.

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Āll data are available either within the manuscript and supplementary material, or directly from the corresponding author.

Competing interests

The authors declare no competing interests.

Funding information

R.V.B. was supported by Breast Cancer Action (charity #1170369-9) and a studentship from the University of Leeds. J.L.T. was supported by a University Academic Fellowship from the University of Leeds. L.M.W. was supported by a fellowship from the Leeds CRUK Cancer Centre.

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Broad, R.V., Jones, S.J., Teske, M.C. et al. Inhibition of interferon-signalling halts cancer-associated fibroblast-dependent protection of breast cancer cells from chemotherapy. Br J Cancer (2021). https://doi.org/10.1038/s41416-020-01226-4

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