Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients

Abstract With the approval of new therapies targeting the PI3K pathway, the detection of PIK3CA mutations has become a key factor in treatment management for HR+/HER2− metastatic breast cancer (MBC). We developed multiplex digital PCR (dPCR) assays to detect and quantify PIK3CA mutations. A first sc...

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Autores principales:	Julien Corné, Fanny Le Du, Véronique Quillien, Florence Godey, Lucie Robert, Héloïse Bourien, Angélique Brunot, Laurence Crouzet, Christophe Perrin, Claudia Lefeuvre-Plesse, Véronique Diéras, Thibault De la Motte Rouge
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/493c25d955e1403bb36fc1a721cea902
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spelling	oai:doaj.org-article:493c25d955e1403bb36fc1a721cea9022021-12-02T16:35:05ZDevelopment of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients10.1038/s41598-021-96644-62045-2322https://doaj.org/article/493c25d955e1403bb36fc1a721cea9022021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-96644-6https://doaj.org/toc/2045-2322Abstract With the approval of new therapies targeting the PI3K pathway, the detection of PIK3CA mutations has become a key factor in treatment management for HR+/HER2− metastatic breast cancer (MBC). We developed multiplex digital PCR (dPCR) assays to detect and quantify PIK3CA mutations. A first screening assay allows the detection of 21 mutations, with a drop-off system targeting the 542–546 hotspot mutations combined with the simultaneous detection of N345K, C420R, H1047L and H1047R mutations. In the case of a positive result, a sequential strategy based on other assays that we have developped allows for precise mutation identification. Clinical validity was determined by analyzing plasma circulating free DNA (cfDNA) from 213 HR+/HER2− MBC samples, as well as DNA extracted from 97 available matched tumors from 89 patients. Our assays have shown reliable specificity, accuracy and reproducibility, with limits of blank of three and four droplets for the screening assay. Sixty-eight patients (32%) had at least one PIK3CA mutation detectable in their plasma, and we obtained 83.1% agreement between the cfDNA analysis and the corresponding tumors. The high sensitivity and robustness of these new dPCR assays make them well-suited for rapid and cost-effective detection of PIK3CA mutations in the plasma of MBC patients.Julien CornéFanny Le DuVéronique QuillienFlorence GodeyLucie RobertHéloïse BourienAngélique BrunotLaurence CrouzetChristophe PerrinClaudia Lefeuvre-PlesseVéronique DiérasThibault De la Motte RougeNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Julien Corné Fanny Le Du Véronique Quillien Florence Godey Lucie Robert Héloïse Bourien Angélique Brunot Laurence Crouzet Christophe Perrin Claudia Lefeuvre-Plesse Véronique Diéras Thibault De la Motte Rouge Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
description	Abstract With the approval of new therapies targeting the PI3K pathway, the detection of PIK3CA mutations has become a key factor in treatment management for HR+/HER2− metastatic breast cancer (MBC). We developed multiplex digital PCR (dPCR) assays to detect and quantify PIK3CA mutations. A first screening assay allows the detection of 21 mutations, with a drop-off system targeting the 542–546 hotspot mutations combined with the simultaneous detection of N345K, C420R, H1047L and H1047R mutations. In the case of a positive result, a sequential strategy based on other assays that we have developped allows for precise mutation identification. Clinical validity was determined by analyzing plasma circulating free DNA (cfDNA) from 213 HR+/HER2− MBC samples, as well as DNA extracted from 97 available matched tumors from 89 patients. Our assays have shown reliable specificity, accuracy and reproducibility, with limits of blank of three and four droplets for the screening assay. Sixty-eight patients (32%) had at least one PIK3CA mutation detectable in their plasma, and we obtained 83.1% agreement between the cfDNA analysis and the corresponding tumors. The high sensitivity and robustness of these new dPCR assays make them well-suited for rapid and cost-effective detection of PIK3CA mutations in the plasma of MBC patients.
format	article
author	Julien Corné Fanny Le Du Véronique Quillien Florence Godey Lucie Robert Héloïse Bourien Angélique Brunot Laurence Crouzet Christophe Perrin Claudia Lefeuvre-Plesse Véronique Diéras Thibault De la Motte Rouge
author_facet	Julien Corné Fanny Le Du Véronique Quillien Florence Godey Lucie Robert Héloïse Bourien Angélique Brunot Laurence Crouzet Christophe Perrin Claudia Lefeuvre-Plesse Véronique Diéras Thibault De la Motte Rouge
author_sort	Julien Corné
title	Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
title_short	Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
title_full	Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
title_fullStr	Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
title_full_unstemmed	Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients
title_sort	development of multiplex digital pcr assays for the detection of pik3ca mutations in the plasma of metastatic breast cancer patients
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/493c25d955e1403bb36fc1a721cea902
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Development of multiplex digital PCR assays for the detection of PIK3CA mutations in the plasma of metastatic breast cancer patients

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