Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.

<h4>Purpose</h4>Decades of quality control efforts have raised the standards of immunohistochemistry (IHC), the principle method used for biomarker testing in breast cancer; however, computational pathology and reverse transcription quantitative PCR (RT-qPCR) may also hold promise for ad...

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Autores principales:	Emanuele Caselli, Cristina Pelliccia, Valeria Teti, Guido Bellezza, Martina Mandarano, Ivana Ferri, Kerstin Hartmann, Mark Laible, Ugur Sahin, Zsuzsanna Varga, Chiara Lupi, Fabrizio Stracci, Angelo Sidoni
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/068c088faafb4f7eba579af269baa966
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spelling	oai:doaj.org-article:068c088faafb4f7eba579af269baa9662021-12-02T20:08:04ZLooking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.1932-620310.1371/journal.pone.0255580https://doaj.org/article/068c088faafb4f7eba579af269baa9662021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0255580https://doaj.org/toc/1932-6203<h4>Purpose</h4>Decades of quality control efforts have raised the standards of immunohistochemistry (IHC), the principle method used for biomarker testing in breast cancer; however, computational pathology and reverse transcription quantitative PCR (RT-qPCR) may also hold promise for additional substantial improvements.<h4>Methods</h4>Herein, we investigated discrepancies in the assessment of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and marker of proliferation Ki67 comparing routinely obtained IHC (and FISH) data (ORI) with the results of manual (REV) and semi-automated (DIA) re-evaluation of the original IHC slides and then with RNA expression data from the same tissue block using the MammaTyper® (MT) gene expression assay.<h4>Results</h4>Correlation for ER and PR was high between ORI IHC and the other three study methods (REV, DIA and RT-qPCR). For HER2, 10 out of 96 discrepant cases can be detected between ORI and REV that involved at least one call in the equivocal category (except for one case). For Ki67, 22 (29.1%) cases were categorized differently by either REV alone (n = 17), DIA alone (n = 15) or both (n = 10) and 28 cases (29.2%) for RT-qPCR. Most of the discrepant Ki67 cases changed from low to high between the original and following assessment and belonged to the intermediate Ki67 expression range (between 9 and 30%).<h4>Conclusions</h4>Determination of the breast cancer biomarkers ER, PR, HER2 and Ki67 at the mRNA level shows high degree of correlation with IHC and compares well with correlations between original with subsequent independent manual or semi-automated IHC assessments. The use of methods with wider dynamic range and higher reproducibility such as RT-qPCR may offer more precise assessment of endocrine responsiveness, improve Ki67 standardization and help resolve HER2 cases that remain equivocal or ambiguous by IHC/FISH. In summary, our findings seem to configure RT-qPCR as a complementary method to be used in cases of either equivocal results or presenting, at the traditional determination assays, biomarkers expressions close to the cut-off values.Emanuele CaselliCristina PellicciaValeria TetiGuido BellezzaMartina MandaranoIvana FerriKerstin HartmannMark LaibleUgur SahinZsuzsanna VargaChiara LupiFabrizio StracciAngelo SidoniPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 9, p e0255580 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Emanuele Caselli Cristina Pelliccia Valeria Teti Guido Bellezza Martina Mandarano Ivana Ferri Kerstin Hartmann Mark Laible Ugur Sahin Zsuzsanna Varga Chiara Lupi Fabrizio Stracci Angelo Sidoni Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
description	<h4>Purpose</h4>Decades of quality control efforts have raised the standards of immunohistochemistry (IHC), the principle method used for biomarker testing in breast cancer; however, computational pathology and reverse transcription quantitative PCR (RT-qPCR) may also hold promise for additional substantial improvements.<h4>Methods</h4>Herein, we investigated discrepancies in the assessment of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2) and marker of proliferation Ki67 comparing routinely obtained IHC (and FISH) data (ORI) with the results of manual (REV) and semi-automated (DIA) re-evaluation of the original IHC slides and then with RNA expression data from the same tissue block using the MammaTyper® (MT) gene expression assay.<h4>Results</h4>Correlation for ER and PR was high between ORI IHC and the other three study methods (REV, DIA and RT-qPCR). For HER2, 10 out of 96 discrepant cases can be detected between ORI and REV that involved at least one call in the equivocal category (except for one case). For Ki67, 22 (29.1%) cases were categorized differently by either REV alone (n = 17), DIA alone (n = 15) or both (n = 10) and 28 cases (29.2%) for RT-qPCR. Most of the discrepant Ki67 cases changed from low to high between the original and following assessment and belonged to the intermediate Ki67 expression range (between 9 and 30%).<h4>Conclusions</h4>Determination of the breast cancer biomarkers ER, PR, HER2 and Ki67 at the mRNA level shows high degree of correlation with IHC and compares well with correlations between original with subsequent independent manual or semi-automated IHC assessments. The use of methods with wider dynamic range and higher reproducibility such as RT-qPCR may offer more precise assessment of endocrine responsiveness, improve Ki67 standardization and help resolve HER2 cases that remain equivocal or ambiguous by IHC/FISH. In summary, our findings seem to configure RT-qPCR as a complementary method to be used in cases of either equivocal results or presenting, at the traditional determination assays, biomarkers expressions close to the cut-off values.
format	article
author	Emanuele Caselli Cristina Pelliccia Valeria Teti Guido Bellezza Martina Mandarano Ivana Ferri Kerstin Hartmann Mark Laible Ugur Sahin Zsuzsanna Varga Chiara Lupi Fabrizio Stracci Angelo Sidoni
author_facet	Emanuele Caselli Cristina Pelliccia Valeria Teti Guido Bellezza Martina Mandarano Ivana Ferri Kerstin Hartmann Mark Laible Ugur Sahin Zsuzsanna Varga Chiara Lupi Fabrizio Stracci Angelo Sidoni
author_sort	Emanuele Caselli
title	Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
title_short	Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
title_full	Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
title_fullStr	Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
title_full_unstemmed	Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.
title_sort	looking for more reliable biomarkers in breast cancer: comparison between routine methods and rt-qpcr.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/068c088faafb4f7eba579af269baa966
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Looking for more reliable biomarkers in breast cancer: Comparison between routine methods and RT-qPCR.

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