Open AccessBarcode sequencing identifies resistant mechanisms to epidermal growth factor receptor inhibitors in circulating tumor DNA of lung cancer patients
Author(s) -
Kitazono Satoru,
Sakai Kazuko,
Yanagitani Noriko,
Ariyasu Ryo,
Yoshizawa Takahiro,
Dotsu Yosuke,
Koyama Junji,
Saiki Masafumi,
Sonoda Tomoaki,
Nishikawa Shingo,
Uchibori Ken,
Horiike Atsushi,
Nishio Kazuto,
Nishio Makoto
Publication year - 2019
Publication title -
cancer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 1347-9032
DOI - 10.1111/cas.14153
Subject(s) - kras , amplicon , epidermal growth factor receptor , digital polymerase chain reaction , t790m , biology , lung cancer , mutation , dna sequencing , cold pcr , amplicon sequencing , resistance mutation , microbiology and biotechnology , barcode , cancer research , polymerase chain reaction , cancer , gene , genetics , medicine , point mutation , reverse transcriptase , 16s ribosomal rna , computer science , operating system
Most patients with epidermal growth factor receptor ( EGFR ) mutation‐positive non‐small cell lung cancer ( NSCLC ) will inevitably develop acquired resistance induced by treatment with EGFR tyrosine kinase inhibitors ( EGFR ‐ TKI ). The mechanisms of resistance to EGFR ‐ TKI are multifactorial, and the detection of these mechanisms is critical for treatment choices in patients who have progressed after EGFR ‐ TKI therapy. We evaluated the feasibility of a molecular barcode method using next‐generation sequencing to detect multifactorial resistance mechanisms in circulating tumor DNA and compared the results with those obtained using other technologies. Plasma samples were collected from 25 EGFR mutation‐positive NSCLC patients after the development of EGFR ‐ TKI resistance. Somatic mutation profiles of these samples were assessed using two methods of next‐generation sequencing and droplet digital PCR (dd PCR ). The positive rate for EGFR ‐sensitizing mutations was 18/25 (72.0%) using dd PCR , 17/25 (68.0%) using amplicon sequencing, and 19/25 (76.0%) using molecular barcode sequencing. Rate of the EGFR T790M resistance mutation among patients with EGFR ‐sensitizing mutations was shown to be 7/18 (38.9%) using dd PCR , 6/17 (35.3%) using amplicon sequencing, and 8/19 (42.1%) using molecular barcode sequencing. Copy number gain in the MET gene was detected in three cases using dd PCR . PIK 3 CA , KRAS and TP 53 mutations were detected using amplicon sequencing. Molecular barcode sequencing detected PIK 3 CA , TP 53 , KRAS , and MAP 2K1 mutations. Results of the three assays were comparable; however, in cell‐free DNA , molecular barcode sequencing detected mutations causing multifactorial resistance more sensitively than did the other assays.