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Inflammatory breast cancer cells are characterized by abrogated TGFβ1-dependent cell motility and SMAD3 activity

  • Preclinical Study
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Abstract

Purpose

Inflammatory breast cancer (IBC) is an aggressive form of breast cancer with elevated metastatic potential, characterized by tumor emboli in dermal and parenchymal lymph vessels. This study has investigated the hypothesis that TGFβ signaling is implicated in the molecular biology of IBC.

Methods

TGFβ1-induced cell motility and gene expression patterns were investigated in three IBC and three non-IBC (nIBC) cell lines. Tissue samples from IBC and nIBC patients were investigated for the expression of nuclear SMAD2, SMAD3, and SMAD4. SMAD protein levels were related to gene expression data.

Results

TGFβ1-induced cell motility was strongly abrogated in IBC cells (P = 0.003). Genes differentially expressed between IBC and nIBC cells post TGFβ1 exposure revealed attenuated expression of SMAD3 transcriptional regulators, but overexpression of MYC target genes in IBC. IBC patient samples demonstrated a near absence of SMAD3 and -4 expression in the primary tumor compared to nIBC patient samples (P < 0.001) and a further reduction of staining intensity in tumor emboli. Integration of gene and protein expression data revealed that a substantial fraction of the IBC signature genes correlated with SMAD3 and these genes are indicative of attenuated SMAD3 signaling in IBC.

Conclusion

We demonstrate attenuated SMAD3 transcriptional activity and SMAD protein expression in IBC, together with obliterated TGFβ1-induced IBC cell motility. The further reduction of nuclear SMAD expression levels in tumor emboli suggests that the activity of these transcription factors is involved in the metastatic dissemination of IBC cells, possibly by enabling collective invasion after partial EMT.

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Data availability

All data generated or analyzed during this study are included in this published article and its supplementary information files. The Affymetrix gene expression datasets E-MTAB-1006 and E-MTAB-1547 are publicly available at https://www.ebi.ac.uk/.

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Acknowledgements

The authors would like to thank Prof. Diether Lambrechts and his research team for their collaboration on RNA-sequencing, and Dr. Cecile Colpaert and Dr. Gert Van den Eynden for their input in the histological analysis. We also thank the World IBC Consortium (WIBCC) for their support.

Funding

This work was supported by an Emmanuel Van der Schueren grant for Charlotte Rypens from Kom op tegen Kanker (Stand up to Cancer), the Flemish cancer society.

Author information

Author notes

  1. Charlotte Rypens and Melike Marsan share first authorship.

Authors and Affiliations

  1. Center for Oncological Research (CORE), Faculty of Medicine and Health Sciences - University of Antwerp, Campus Drie Eiken - Universiteitsplein 1, 2610, Wilrijk - Antwerp, Belgium

    Charlotte Rypens, Christophe Van Berckelaer, Charlotte Billiet, Peter van Dam, Piet Dirix, Peter Vermeulen & Steven J. Van Laere

  2. Translational Cancer Research Unit (TCRU), GZA Ziekenhuizen campus Sint-Augustinus, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium

    Charlotte Rypens, Melike Marsan, Christophe Van Berckelaer, Peter Vermeulen, Luc Dirix & Steven J. Van Laere

  3. Multidisciplinary Breast Clinic, Unit Gynaecologic Oncology, Antwerp University Hospital (UZA), Antwerp, Belgium

    Christophe Van Berckelaer & Peter van Dam

  4. Radiation Oncology, Iridium Kankernetwerk, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium

    Charlotte Billiet & Piet Dirix

  5. Karel de Grote Hogeschool, Antwerp, Belgium

    Kirsten Melis

  6. University Complutense, Madrid, Spain

    Sara Perez Lopez

  7. Division of Surgical Sciences, Department of Surgery, Duke University Medical Sciences, Durham, NC, USA

    Gayathri R. Devi

  8. Predictive Oncology Team, Centre de Recherche en Cancérologie de Marseille (CRCM), Inserm, CNRS, Aix-Marseille Université, Institut Paoli-Calmettes, Marseille, France

    Pascal Finetti & François Bertucci

  9. Morgan Welch Inflammatory Breast Cancer Research Program and Clinic & Department of Breast Medical Oncology, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA

    Naoto T. Ueno

  10. Department of Oncology, KU Leuven & Department of Gynaecology and Obstetrics, University Hospitals Leuven, Leuven, Belgium

    Patrick Neven

  11. GZA Ziekenhuizen, campus Sint-Augustinus, Oosterveldlaan 24, 2610, Wilrijk - Antwerp, Belgium

    Luc Dirix

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  1. Charlotte Rypens
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Corresponding author

Correspondence to Charlotte Rypens.

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The authors declare that they have no conflict of interest.

Ethical approval

This study is conducted in accordance with the ethical standards of the GZA Hospitals.

Research involved human and animal participants

This article does not contain any studies with animals performed by any of the authors.

Informed consent

The need for informed consent was waived by the ethical committee because remnant, non-diagnostic tissue was used for all analyses. Patient have had the opportunity to opt-out.

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Rypens, C., Marsan, M., Van Berckelaer, C. et al. Inflammatory breast cancer cells are characterized by abrogated TGFβ1-dependent cell motility and SMAD3 activity. Breast Cancer Res Treat 180, 385–395 (2020). https://doi.org/10.1007/s10549-020-05571-z

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