Skip to main content
Top
Published in: Virchows Archiv 3/2019

01-09-2019 | Breast Cancer | Original Article

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

Authors: Fabio Aimi, Maria-Giuseppina Procopio, Maria Teresa Alvarez Flores, Jean-Philippe Brouland, Nathalie Piazzon, Saška Brajkovic, Diego Gabriel Dupouy, Martin Gijs, Laurence de Leval

Published in: Virchows Archiv | Issue 3/2019

Login to get access

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.
Appendix
Available only for authorised users
Literature
1.
go back to reference McGuire A, Brown J, Malone C, McLaughlin R, Kerin M (2015) Effects of age on the detection and management of breast cancer. Cancers (Basel) 7(2):908–929CrossRef McGuire A, Brown J, Malone C, McLaughlin R, Kerin M (2015) Effects of age on the detection and management of breast cancer. Cancers (Basel) 7(2):908–929CrossRef
2.
go back to reference Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136(5):E359–E386CrossRefPubMed Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D, Bray F (2015) Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer 136(5):E359–E386CrossRefPubMed
3.
go back to reference Banin Hirata BK, Oda JM, Losi Guembarovski R, Ariza CB, de Oliveira CE, Watanabe MA (2014) Molecular markers for breast cancer: prediction on tumor behavior. Dis Markers 2014:513158CrossRefPubMedPubMedCentral Banin Hirata BK, Oda JM, Losi Guembarovski R, Ariza CB, de Oliveira CE, Watanabe MA (2014) Molecular markers for breast cancer: prediction on tumor behavior. Dis Markers 2014:513158CrossRefPubMedPubMedCentral
5.
go back to reference Bandyopadhyay S, Bluth MH, Ali-Fehmi R (2018) Breast carcinoma: updates in molecular profiling 2018. Clin Lab Med 38(2):401–420CrossRefPubMed Bandyopadhyay S, Bluth MH, Ali-Fehmi R (2018) Breast carcinoma: updates in molecular profiling 2018. Clin Lab Med 38(2):401–420CrossRefPubMed
6.
go back to reference Ciftlik AT, Lehr H-A, Gijs MAM (2013) Microfluidic processor allows rapid HER2 immunohistochemistry of breast carcinomas and significantly reduces ambiguous (2+) read-outs. Proc Natl Acad Sci 110(14):5363–5368CrossRefPubMedPubMedCentral Ciftlik AT, Lehr H-A, Gijs MAM (2013) Microfluidic processor allows rapid HER2 immunohistochemistry of breast carcinomas and significantly reduces ambiguous (2+) read-outs. Proc Natl Acad Sci 110(14):5363–5368CrossRefPubMedPubMedCentral
7.
go back to reference Dupouy DG et al (2016) Continuous quantification of HER2 expression by microfluidic precision immunofluorescence estimates HER2 gene amplification in breast cancer. Sci Rep 6 Dupouy DG et al (2016) Continuous quantification of HER2 expression by microfluidic precision immunofluorescence estimates HER2 gene amplification in breast cancer. Sci Rep 6
8.
go back to reference Brajkovic S, Dupouy DG, De Leval L, Gijs MAM (2017) Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections. Lab Investig 97(8):983–991CrossRefPubMed Brajkovic S, Dupouy DG, De Leval L, Gijs MAM (2017) Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections. Lab Investig 97(8):983–991CrossRefPubMed
9.
go back to reference Hammond MEH et al (2010) American Society of Clinical Oncology/College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Clin Oncol 28(16):2784–2795CrossRefPubMedPubMedCentral Hammond MEH et al (2010) American Society of Clinical Oncology/College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Clin Oncol 28(16):2784–2795CrossRefPubMedPubMedCentral
10.
go back to reference Wolff AC, Hammond ME, Hicks DG, Dowsett M, McShane L, Allison KH, Allred DC, Bartlett JM, Bilous M, Fitzgibbons P, Hanna W, Jenkins RB, Mangu PB, Paik S, Perez EA, Press MF, Spears PA, Vance GH, Viale G, Hayes DF, American Society of Clinical Oncology, College of American Pathologists (2013) Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. TL - 31. J Clin Oncol 31(31):3997–4013CrossRefPubMed Wolff AC, Hammond ME, Hicks DG, Dowsett M, McShane L, Allison KH, Allred DC, Bartlett JM, Bilous M, Fitzgibbons P, Hanna W, Jenkins RB, Mangu PB, Paik S, Perez EA, Press MF, Spears PA, Vance GH, Viale G, Hayes DF, American Society of Clinical Oncology, College of American Pathologists (2013) Recommendations for human epidermal growth factor receptor 2 testing in breast cancer: American Society of Clinical Oncology/College of American Pathologists clinical practice guideline update. TL - 31. J Clin Oncol 31(31):3997–4013CrossRefPubMed
11.
go back to reference Coates AS et al (2015) Tailoring therapies-improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015. Ann Oncol Coates AS et al (2015) Tailoring therapies-improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015. Ann Oncol
12.
go back to reference Nguyen HT, Trouillon R, Matsuoka S, Fiche M, de Leval L, Bisig B, Gijs MA (2017) Microfluidics-assisted fluorescence in situ hybridization for advantageous human epidermal growth factor receptor 2 assessment in breast cancer. Lab Investig 97(1):93–103CrossRefPubMed Nguyen HT, Trouillon R, Matsuoka S, Fiche M, de Leval L, Bisig B, Gijs MA (2017) Microfluidics-assisted fluorescence in situ hybridization for advantageous human epidermal growth factor receptor 2 assessment in breast cancer. Lab Investig 97(1):93–103CrossRefPubMed
13.
go back to reference Troxell ML, Long T, Hornick JL, Ambaye AB, Jensen KC (2017) Comparison of estrogen and progesterone receptor antibody reagents using proficiency testing data. Arch Pathol Lab Med 141(10):1402–1412CrossRefPubMed Troxell ML, Long T, Hornick JL, Ambaye AB, Jensen KC (2017) Comparison of estrogen and progesterone receptor antibody reagents using proficiency testing data. Arch Pathol Lab Med 141(10):1402–1412CrossRefPubMed
14.
go back to reference Calhoun BC, Mosteller B, Warren D, Smith M, Jordi Rowe J, Lanigan CP, Mrazeck KC, Walker E, Newell AH, Jones R (2018) Analytical and clinical performance of progesterone receptor antibodies in breast cancer. Ann Diagn Pathol 35:21–26CrossRefPubMed Calhoun BC, Mosteller B, Warren D, Smith M, Jordi Rowe J, Lanigan CP, Mrazeck KC, Walker E, Newell AH, Jones R (2018) Analytical and clinical performance of progesterone receptor antibodies in breast cancer. Ann Diagn Pathol 35:21–26CrossRefPubMed
15.
go back to reference Roepman P, Horlings HM, Krijgsman O, Kok M, Bueno-de-Mesquita JM, Bender R, Linn SC, Glas AM, van de Vijver M (2009) Microarray-based determination of estrogen receptor, progesterone receptor, and HER2 receptor status in breast cancer. Clin Cancer Res 15(22):7003–7011CrossRefPubMed Roepman P, Horlings HM, Krijgsman O, Kok M, Bueno-de-Mesquita JM, Bender R, Linn SC, Glas AM, van de Vijver M (2009) Microarray-based determination of estrogen receptor, progesterone receptor, and HER2 receptor status in breast cancer. Clin Cancer Res 15(22):7003–7011CrossRefPubMed
16.
go back to reference Prat A et al (2013) Prognostic significance of progesterone receptor-positive tumor cells within immunohistochemically defined luminal a breast cancer. J Clin Oncol Prat A et al (2013) Prognostic significance of progesterone receptor-positive tumor cells within immunohistochemically defined luminal a breast cancer. J Clin Oncol
17.
go back to reference Goldhirsch A, Winer EP, Coates AS, Gelber RD, Piccart-Gebhart M, Thürlimann B, Senn HJ, Albain KS, André F, Bergh J, Bonnefoi H, Bretel-Morales D, Burstein H, Cardoso F, Castiglione-Gertsch M, Coates AS, Colleoni M, Costa A, Curigliano G, Davidson NE, di Leo A, Ejlertsen B, Forbes JF, Gelber RD, Gnant M, Goldhirsch A, Goodwin P, Goss PE, Harris JR, Hayes DF, Hudis CA, Ingle JN, Jassem J, Jiang Z, Karlsson P, Loibl S, Morrow M, Namer M, Kent Osborne C, Partridge AH, Penault-Llorca F, Perou CM, Piccart-Gebhart MJ, Pritchard KI, Rutgers EJT, Sedlmayer F, Semiglazov V, Shao ZM, Smith I, Thürlimann B, Toi M, Tutt A, Untch M, Viale G, Watanabe T, Wilcken N, Winer EP, Wood WC (2013) Personalizing the treatment of women with early breast cancer: highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Ann Oncol 24:2206–2223CrossRefPubMedPubMedCentral Goldhirsch A, Winer EP, Coates AS, Gelber RD, Piccart-Gebhart M, Thürlimann B, Senn HJ, Albain KS, André F, Bergh J, Bonnefoi H, Bretel-Morales D, Burstein H, Cardoso F, Castiglione-Gertsch M, Coates AS, Colleoni M, Costa A, Curigliano G, Davidson NE, di Leo A, Ejlertsen B, Forbes JF, Gelber RD, Gnant M, Goldhirsch A, Goodwin P, Goss PE, Harris JR, Hayes DF, Hudis CA, Ingle JN, Jassem J, Jiang Z, Karlsson P, Loibl S, Morrow M, Namer M, Kent Osborne C, Partridge AH, Penault-Llorca F, Perou CM, Piccart-Gebhart MJ, Pritchard KI, Rutgers EJT, Sedlmayer F, Semiglazov V, Shao ZM, Smith I, Thürlimann B, Toi M, Tutt A, Untch M, Viale G, Watanabe T, Wilcken N, Winer EP, Wood WC (2013) Personalizing the treatment of women with early breast cancer: highlights of the St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2013. Ann Oncol 24:2206–2223CrossRefPubMedPubMedCentral
Metadata
Title
Microfluidic-based immunohistochemistry for breast cancer diagnosis: a comparative clinical study
Authors
Fabio Aimi
Maria-Giuseppina Procopio
Maria Teresa Alvarez Flores
Jean-Philippe Brouland
Nathalie Piazzon
Saška Brajkovic
Diego Gabriel Dupouy
Martin Gijs
Laurence de Leval
Publication date
01-09-2019
Publisher
Springer Berlin Heidelberg
Published in
Virchows Archiv / Issue 3/2019
Print ISSN: 0945-6317
Electronic ISSN: 1432-2307
DOI
https://doi.org/10.1007/s00428-019-02616-7

Other articles of this Issue 3/2019

Virchows Archiv 3/2019 Go to the issue