A Multicenter Study to Assess EGFR Mutational Status in Plasma: Focus on an Optimized Workflow for Liquid Biopsy in a Clinical Setting

Cancers

卷:10

Daniëlla Galdermans¹  Julie Jacobs²  Elien Augustus²  Laure Sorber²  Patrick Pauwels²  Kaat Van Casteren²  Christian Rolfo²  Karen Zwaenepoel²  Ulrike Himpe³  Thierry Pieters³  XiangHua Zhang⁴  Koen De Winne⁵  Paul Germonpré⁶  Anneke Lefebure⁷  Els De Droogh⁷  Koen Deschepper⁸  Ingel Demedts⁹  JanP. Van Meerbeeck¹⁰  Frédérique Bustin¹⁰  Erika Saenen¹⁰  Ann-Marie Morel¹⁰  Sofie Derijcke¹⁰ 

[1] Thoracic Oncology, Antwerp University Hospital (UZA), 2650 Edegem, Belgium;

[2] Center for Oncological Research Antwerp (CORE), University of Antwerp, 2610 Wilrijk, Belgium;

[3] Department of Pulmonary diseases, AZ Delta, 8800 Roeselare, Belgium;

[4] Department of Pulmonology &

[5] Laboratory of Pathological Anatomy, Antwerp University Hospital (UZA), 2650 Edegem, Belgium;

[6] Pneumology, Cliniques Universitaires Saint-Luc, 1200 Bruxelles, Belgium;

[7] Pneumology, ZNA Middelheim, 2020 Antwerp, Belgium;

[8] Pneumology-Thoracic Oncology, AZ Groeninge, 8500 Kortrijk, Belgium;

[9] Pneumology/Allergology, CHR de la Citadelle, 4000 Liège, Belgium;

[10] Thoracic Oncology Group Antwerp (TOGA), University of Antwerp, 2610 Wilrijk, Belgium;

关键词: liquid biopsy;    non-small cell lung cancer (NSCLC);    EGFR;    ctDNA;    ddPCR;   

DOI  :  10.3390/cancers10090290

来源: DOAJ

【 摘 要 】

A multicenter study was performed to determine an optimal workflow for liquid biopsy in a clinical setting. In total, 549 plasma samples from 234 non-small cell lung cancer (NSCLC) patients were collected. Epidermal Growth Factor Receptor (EGFR) circulating cell-free tumor DNA (ctDNA) mutational analysis was performed using digital droplet PCR (ddPCR). The influence of (pre-) analytical variables on ctDNA analysis was investigated. Sensitivity of ctDNA analysis was influenced by an interplay between increased plasma volume (p < 0.001) and short transit time (p = 0.018). Multistep, high-speed centrifugation both increased plasma generation (p < 0.001) and reduced genomic DNA (gDNA) contamination. Longer transit time increased the risk of hemolysis (p < 0.001) and low temperatures were shown to have a negative effect. Metastatic sites were found to be strongly associated with ctDNA detection (p < 0.001), as well as allele frequency (p = 0.034). Activating mutations were detected in a higher concentration and allele frequency compared to the T790M mutation (p = 0.003, and p = 0.002, respectively). Optimization of (pre-) analytical variables is key to successful ctDNA analysis. Sufficient plasma volumes without hemolysis or gDNA contamination can be achieved by using multistep, high-speed centrifugation, coupled with short transit time and temperature regulation. Metastatic site location influenced ctDNA detection. Finally, ctDNA levels might have further value in detecting resistance mechanisms.

【 授权许可】

Unknown