Plasma-based analysis of ERBB2 mutational status by multiplex digital PCR in a large series of patients with metastatic breast cancer

Sep 21, 2026

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Erb-b2 receptor tyrosine kinase 2 (ERBB2)-activating mutations are actionable alterations across cancers, including metastatic breast cancer. Multiplex digital PCR assays were developed to detect and quantify ERBB2 mutations in circulating tumor DNA from liquid biopsies. Plasma from 272 HR+/HER2− metastatic breast cancer patients was analyzed for 17 ERBB2 mutations using a screening assay on the Crystal dPCR™ naica® platform. Nine patients (3.3%) carried at least one mutation, with higher rates in lobular histology (5.9%) versus IBC-NST (2.6%).

LanguageEnglish

CategoryResearch & Report

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Pages16

Reading Time40 min

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Introduction

Development of multiplex digital PCR assay

ERBB2 mutational detection in patient plasma

Mutation frequencies and concordance with tumor samples

Plasma-based analysis of ERBB2 mutational status by multiplex digital PCR in a large series of patients with metastatic breast cancer Julien Corn´e1 , V´eronique Quillien 1,2,3, Florence Godey 1,2,3, Mathilde Cherel 1, Agathe Cochet2, Fanny Le Du2, Lucie Robert2, H´eloı¨se Bourien2, Ang´elique Brunot2, Laurence Crouzet2, Christophe Perrin2, Claudia Lefeuvre-Plesse2, V´eronique Di´eras2 and Thibault De la Motte Rouge2 1 Department of Biology, Centre Eug`ene Marquis, Unicancer, Rennes, France 2 Department of Medical Oncology, Centre Eug`ene Marquis, Unicancer, Rennes, France 3 INSERM U1242, University of Rennes, France Keywords circulating tumor DNA; ERBB2; liquid biopsies; metastatic breast cancer; multiplex digital PCR; plasma Correspondence J. Corn´e, Centre Eug`ene Marquis, Avenue de la Bataille Flandres Dunkerque, CS 44229, 35042 Rennes, France Fax: +33 299253164 Tel: +33 299253192 E-mail: [j.corne@rennes.unicancer.fr](j.corne@rennes.unicancer.fr) (Received 7 August 2023, revised 16 November 2023, accepted 15 January 2024, available online 29 January 2024) doi:10 . 1002/1878-0261 .13592 Erb-b2 receptor tyrosine kinase 2 (ERBB2)-activating mutations are therapeutically actionable alterations found in various cancers, including metastatic breast cancer (MBC) . We developed multiplex digital PCR assays to detect and quantify ERBB2 mutations in circulating tumor DNA from liquid biopsies. We studied the plasma from 272 patients with hormone-receptorpositive, human epidermal growth factor receptor 2-negative (HR+/HER2􀀁) MBC to detect 17 ERBB2 mutations using a screening assay. The assay was developed on the three-color Crystal dPCR™ naica® platform with a twostep strategy for precise mutation identiﬁcation. We found that nine patients (3 . 3%) harbored at least one ERBB2 mutation. The mutation rate was higher in patients with lobular histology (5 .9%) compared to invasive breast carcinoma of no special type (2 .6%) . A total of 12 mutations were found with the following frequencies: L755S (25.00%), V777L (25.00%), S310Y (16.67%), L869R (16 .67%), S310F (8 . 33%), and D769H (8 . 33%) . Matched tumor samples from six patients identiﬁed the same mutations with an 83% concordance rate. In summary, our highly sensitive multiplex digital PCR assays are well suited for plasma-based monitoring of ERBB2 mutational status inpatients with MBC. 1. Introduction Human epidermal growth factor receptor 2 (HER2) is encoded by the Erb-b2 receptor tyrosine kinase 2 (ERBB2) gene. HER2 is a common oncogenic driver in breast cancer and a major drug target. Most deregulation of HER2 signaling is caused by HER2/ERBB2 ampliﬁcation but activating mutations leading to constitutive HER2 signaling can also occur, both in patients with hormone receptor (HR)-positive, HER2-negative or HR+/HER2+ metastatic breast cancer (MBC), with different mutation landscapes Abbreviations cfDNA, circulating cell-free DNA; COSMIC, Catalogue of Somatic Mutations in Cancer; ctDNA, circulating tumor DNA; CV, coefﬁcient of variation; DO, drop-off; dPCR, digital PCR; ER, estrogen receptor; ERBB2, Erb-b2 receptor tyrosine kinase 2; ESCAT, ESMO Scale for Clinical Actionability of Molecular Targets; ET, endocrine therapy; FFPE, formalin-ﬁxed, parafﬁn-embedded; gBlock, gBlock gene fragments; gDNA, genomic DNA; HER2, human epidermal growth factor receptor 2; HGVS, Human Genome Variation Society; HR, hormone receptor; IBC-NST, invasive breast carcinoma of no special type; IDC, invasive ductal carcinoma; ILC, invasive lobular carcinoma; LOB, limit of blank; LOD, limit of detection; MAF, mutant allelic frequency; MBC, metastatic breast cancer; MUT, mutated; NF1, neuroﬁbromin 1; NGS, nextgeneration sequencing; OS, overall survival; PIK3CA, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha; REF, reference; SD, stable disease; WT, wild-type. 2714 Molecular Oncology 18 (2024) 2714–2729 ª 2024 The Authors. Molecular Oncology publis

FAQs

What was the main purpose of the study?

To develop multiplex digital PCR assays for detecting and quantifying ERBB2 mutations in circulating tumor DNA from plasma liquid biopsies, and to evaluate them in a large metastatic breast cancer patient series.

How many ERBB2 mutations and patients were analyzed?

The study screened 272 metastatic breast cancer patients for 17 ERBB2 mutations using a plasma-based screening assay.

What were the key findings about ERBB2 mutation rates?

ERBB2 mutations were found in 3.3% of patients overall, with a higher mutation rate in lobular histology (5.9%) than in invasive breast carcinoma of no special type (2.6%), and matching tumor samples showed 83% concordance.

Sep 21, 2026

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