Longitudinal Multi-Parametric Liquid Biopsy Approach Identifies Unique Features of Circulating Tumor Cell, Extracellular Vesicle, and Cell-Free DNA Characterization for Disease Monitoring in Metastatic Breast Cancer Patients.

Corinna Keup,Rainer Kimmig,Markus Storbeck,Oliver Hoffmann,Vinay Suryaprakash,Sabine Kasimir-Bauer

doi:10.3390/cells10020212

Corinna Keup, Rainer Kimmig + Show 4 more

Open Access

https://doi.org/10.3390/cells10020212

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Journal: Cells	Publication Date: Jan 21, 2021
Citations: 28	License type: CC BY 4.0

Affiliation: Essen University Hospital, Qiagen (Germany)

Abstract

Dynamics of mRNA from circulating tumor cells (CTCs), mRNA from extracellular vesicles (EVs), and cell-free DNA (cfDNA) were assessed to examine the relevance of a longitudinal multi-parametric liquid biopsy strategy. Eighteen milliliters of blood was drawn from 27 hormone receptor-positive and human epidermal growth factor receptor 2 (HER2)-negative metastatic breast cancer (MBC) patients at disease progression and at two subsequent radiologic staging time points. CTC mRNA and EV mRNA were analyzed using multi-marker qPCR, and cfDNA was analyzed using targeted next-generation sequencing (NGS). The presence of ERBB2 or ERBB3 overexpression signals in CTCs significantly correlated with disease progression (87% specificity, 36% sensitivity, p-value = 0.023), and the presence of either ERBB3 signals in CTCs or EVs or cfDNA variants in ERBB3 also showed a significant association with progressive MBC. Fluctuations during treatment were detected in the EV fraction with the appearance of hitherto undetected ERCC1 signals correlating with progressive disease (97% specificity, 18% sensitivity, p-value = 0.030). Allele frequency development of ESR1 and PIK3CA variants detected at subsequent staging time points could be used as a predictor for therapy success and, importantly, might help guide therapy decisions. The three analytes, each with their own unique features for disease monitoring, were shown to be complementary, underlining the usefulness of the longitudinal multi-parametric liquid biopsy approach.

Highlights

Liquid biopsies provide analytes that are powerful for disease monitoring and for the identification of molecular disease features [1,2]
Patient specimens were only included in this multi-parametric liquid biopsy study if sufficient material existed for matched circulating tumor cells (CTCs) isolation (10 mL whole blood), extracellular vesicles (EVs) isolation (4 mL plasma), cell-free DNA (cfDNA) isolation (≥1 mL plasma from CTC-depleted blood), and reliable sequencing result interpretation (>4 nM library yield, >4 million reads fragments per sample)
As this is a longitudinal study, patient samples were only included if the abovementioned inclusion criteria were fulfilled for all three blood samples drawn at the three consecutive staging time points

Summary

Introduction

Liquid biopsies provide analytes that are powerful for disease monitoring and for the identification of molecular disease features [1,2]. Besides the determination of somatic alterations and disrupted pathways, liquid biopsy testing harbors great potential for sensitive detection of minimal residual disease (MRD) or therapy resistance and, recurrence or disease progression [4]. In breast cancer (BC), the leading cancer in women worldwide [5], an increase in circulating tumor cell (CTC) count [6] and an increase in circulating tumor DNA (ctDNA) concentration have been shown to correlate with disease progression [7,8]. In metastatic BC (MBC) patients, a reduction of the apoptotic CTC count by ≥50% (after one completed treatment cycle) correlated with stable disease, while a reduction of apoptotic. In primary BC, the persistent detection of Cytokeratin-19-positive CTCs during the first 5 years after surgery has been related to an increased risk of late relapse [10]

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