Tag-based next generation sequencing: a feasible and reliable assay for EGFR T790M mutation detection in circulating tumor DNA of non small cell lung cancer patients

• Published in: Molecular medicine (Cambridge, Mass.) Vol. 25; no. 1; pp. 15 - 13
• Main Authors: Dono, Mariella, De Luca, Giuseppa, Lastraioli, Sonia, Anselmi, Giorgia, Dal Bello, Maria Giovanna, Coco, Simona, Vanni, Irene, Grossi, Francesco, Vigani, Antonella, Genova, Carlo, Ferrarini, Manlio, Ravetti, Jean Louis, Zupo, Simona
• Format: Journal Article
• Language: English
• Published: London BioMed Central 27.04.2019; BMC
• Subjects: Biomedical and Life Sciences; Biomedicine; Biopsy; Carcinoma, Non-Small-Cell Lung - genetics; Circulating tumor DNA; Circulating Tumor DNA - genetics; EGFR TKIs; ErbB Receptors - genetics; FDA approval; Female; Gene amplification; High-Throughput Nucleotide Sequencing - methods; Humans; Laboratories; Liquid biopsy; Lung cancer; Lung Neoplasms - genetics; Male; Molecular Medicine; Molecular tag; Mutation; Mutation - genetics; NSCLC; Performance evaluation; Plasma; Real time; Research Article; T790M resistance mutation; Targeted cancer therapy; Values; United States > US; Italy; EGFR TKIs; NSCLC; T790M resistance mutation; Molecular tag; Liquid biopsy; C797S; Next generation sequencing; Circulating tumor DNA
• ISSN: 1076-1551; 1528-3658; 1528-3658
• DOI: 10.1186/s10020-019-0082-5

Background The demonstration of EGFR T790M gene mutation in plasma is crucial to assess the eligibility of Non Small Cell Lung Cancer (NSCLC) patients, who have acquired resistance to first or second generation Tyrosine Kinase Inhibitors (TKIs), to receive a subsequent treatment with osimertinib. Since circulating tumor DNA (ctDNA) is present in very low amounts in plasma, high sensitive and specific methods are required for molecular analysis. Improving sensitivity of T790M mutation detection in plasma ctDNA enables a larger number of NSCLC patients to receive the appropriate therapy without any further invasive procedure. Methods A tag-based next generation sequencing (NGS) platform capable of tagging rare circulating tumor DNA alleles was employed in this study for the identification of T790M mutation in 42 post-TKI NSCLC patients. Results Compared to Real Time PCR, tag-based NGS improved the T790M detection rate (42.85% versus 21.4%, respectively), especially in those cases with a low median mutation abundance (i.e. 0.24, range 0.07–0.78). Moreover, the tag-based NGS identified EGFR activating mutations more efficiently than Real Time PCR (85.7% versus 61.9% detection rate, respectively), particularly of the L858R variant type (0.06–0.75 mutation abundance range). Patients in whom the T790M mutation was detected in plasma, achieved an objective response to osimertinib (9/14, 64.28%). Conclusions Tag-based NGS represents an accurate and sensitive tool in a clinical setting for non-invasive assessment and monitoring of T790M variant in NSCLC patients.