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Microfluidic-based immunohistochemistry for breast cancer diagnosis: a comparative clinical study

Virchows Archiv volume 475, pages 313–323 (2019)Cite this article

Abstract

Breast cancer is a highly heterogeneous disease. The efficacy of tailored therapeutic strategies relies on the precise detection of diagnostic biomarkers by immunohistochemistry (IHC). Therefore, considering the increasing incidence of breast cancer cases, a concomitantly time-efficient and accurate diagnosis is clinically highly relevant. Microfluidics is a promising innovative technology in the field of tissue diagnostic, enabling for rapid, reliable, and automated immunostaining. We previously reported the microfluidic-based HER2 (human epidermal growth factor receptor 2) detection in breast carcinomas to greatly correlate with the HER2 gene amplification level. Here, we aimed to develop a panel of microfluidic-based IHC protocols for prognostic and therapeutic markers routinely assessed for breast cancer diagnosis, namely HER2, estrogen/progesterone receptor (ER/PR), and Ki67 proliferation factor. The microfluidic IHC protocol for each marker was optimized to reach high staining quality comparable to the standard procedure, while concomitantly shortening the staining time to 16 min—excluding deparaffinization and antigen retrieval step—with a turnaround time reduction up to 7 folds. Comparison of the diagnostic score on 50 formaldehyde-fixed paraffin-embedded breast tumor resections by microfluidic versus standard staining showed high concordance (overall agreement: HER2 94%, ER 95.9%, PR 93.6%, Ki67 93.7%) and strong correlation (ρ coefficient: ER 0.89, PR 0.88, Ki67 0.87; p < 0.0001) for all the analyzed markers. Importantly, HER2 genetic reflex test for all discordant cases confirmed the scores obtained by the microfluidic technique. Overall, the microfluidic-based IHC represents a clinically validated equivalent approach to the standard chromogenic staining for rapid, accurate, and automated breast cancer diagnosis.

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Acknowledgments

The authors thank Noguet Véronique and Prod’Hom Nicole for performing BenchMark stainings, Cloé Bregnard and Monica Esteves for performing FISH analysis, and Jean-Daniel Roman for the support provided with digital imaging.

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Authors and Affiliations

  1. Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne, CH-1015, Lausanne, Switzerland

    Fabio Aimi & Martin Gijs

  2. Institute of Pathology and Molecular Pathology, University Hospital Zurich, CH-8091, Zurich, Switzerland

    Maria-Giuseppina Procopio

  3. Institute of Pathology, Centre Hospitalier Universitaire Vaudois, CH-1011, Lausanne, Switzerland

    Maria Teresa Alvarez Flores, Jean-Philippe Brouland, Nathalie Piazzon & Laurence de Leval

  4. Lunaphore Technologies SA, EPFL Innovation Park-Building C, CH-1015, Lausanne, Switzerland

    Saška Brajkovic & Diego Gabriel Dupouy

Authors
  1. Fabio Aimi
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  2. Maria-Giuseppina Procopio
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  3. Maria Teresa Alvarez Flores
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  4. Jean-Philippe Brouland
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  5. Nathalie Piazzon
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  6. Saška Brajkovic
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  7. Diego Gabriel Dupouy
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  8. Martin Gijs
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  9. Laurence de Leval
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Contributions

S.B., D.G.D., M.G., N.P., and L.d.L designed the study. S.B. and N.P. supervised study execution. F.A. ran the experiments. M.T.A.F. and J.P.B. blindly scored the assessed cases. M.G.P. interpreted the results, F.A. and M.G.P designed the figures, and M.G.P and F.A. wrote the paper. All authors contributed to manuscript writing and revision, and they all approved the submitted version. The authors declare that the article is presently not under consideration for publication in another journal and has not been published previously.

Corresponding author

Correspondence to Saška Brajkovic.

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Conflict of interest

At the submission of the paper, DGD and SB are employed by Lunaphore Technologies SA, which is commercializing a microfluidic-based staining device. DGD and MAMG have equity interest in Lunaphore Technologies SA. The other authors declare no conflict of interest.

Ethical approval

A total of 50 FFPE breast cancer tissue specimens were retrieved from the Pathology Institute of the University Hospital of Lausanne (CHUV) between January 2016 and April 2017 with the ethical committee approval (protocol number 511-12).

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Aimi, F., Procopio, MG., Alvarez Flores, M.T. et al. Microfluidic-based immunohistochemistry for breast cancer diagnosis: a comparative clinical study. Virchows Arch 475, 313–323 (2019). https://doi.org/10.1007/s00428-019-02616-7

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  • DOI: https://doi.org/10.1007/s00428-019-02616-7

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