title: Next-generation sequencing of tyrosine kinase inhibitor-resistant non-small-cell lung cancers in patients harboring epidermal growth factor-activating mutations.
abstract: BACKGROUND
  The aim of this study was to detect the epidermal growth factor receptor (EGFR)-activating mutations and other oncogene alterations in patients with non-small-cell lung cancers (NSCLC) who experienced a treatment failure in response to EGFR-tyrosine kinase inhibitors (TKIs) with a next generation sequencer.
  
  
  METHODS
  Fifteen patients with advanced NSCLC previously treated with EGFR-TKIs were examined between August 2005 and October 2014. For each case, new biopsies were performed, followed by DNA sequencing on an Ion Torrent Personal Genome Machine (PGM) system using the Ion AmpliSeq Cancer Hotspot Panel version 2.
  
  
  RESULTS
  All 15 patients were diagnosed with NSCLC harboring EGFR-activating mutations (seven cases of exon 19 deletion, seven cases of L858R in exon 21, and one case of L861Q in exon 21). Of the 15 cases, acquired T790M resistance mutations were detected in 9 (60.0%) patients. In addition, other mutations were identified outside of EGFR, including 13 cases (86.7%) exhibiting TP53 P72R mutations, 5 cases (33.3%) of KDR Q472H, and 2 cases (13.3%) of KIT M541L.
  
  
  CONCLUSIONS
  Here, we showed that next-generation sequencing (NGS) is able to detect EGFR T790M mutations in cases not readily diagnosed by other conventional methods. Significant differences in the degree of EGFR T790M and other EGFR-activating mutations may be indicative of the heterogeneity of disease phenotype evident within these patients. The co-existence of known oncogenic mutations within each of these patients may play a role in acquired EGFR-TKIs resistance, suggesting the need for alternative treatment strategies, with PCR-based NGS playing an important role in disease diagnosis.
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. masago@fbri.org.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Masago
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. sfujita@fbri.org.
authors.forename: Shiro
authors.initials: S
authors.lastname: Fujita
authors.affiliation: Thermo Fisher Scientific, Tokyo, Japan. miho.muraki@thermofisher.com.
authors.forename: Miho
authors.initials: M
authors.lastname: Muraki
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. a-hata@fbri.org.
authors.forename: Akito
authors.initials: A
authors.lastname: Hata
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. chiyuki-okuda@fbri.org.
authors.forename: Chiyuki
authors.initials: C
authors.lastname: Okuda
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. kyokoniceday@hotmail.co.jp.
authors.forename: Kyoko
authors.initials: K
authors.lastname: Otsuka
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. naya@fbri.org.
authors.forename: Reiko
authors.initials: R
authors.lastname: Kaji
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. jumpeinr2tfm3@fbri.org.
authors.forename: Jumpei
authors.initials: J
authors.lastname: Takeshita
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. katou@fbri.org.
authors.forename: Ryoji
authors.initials: R
authors.lastname: Kato
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. katakami@fbri.org.
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Katakami
authors.affiliation: Division of Integrated Oncology, Institute of Biomedical Research and Innovation, 2-2 Minatojima-minamimachi, Cyuo-ku, Kobe City, Hyogo, 650-0047, Japan. yhirata@fbri.org.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Hirata
mesh_terms: D000230:Adenocarcinoma / Q000188:drug therapy / Q000235:genetics*; D000368:Aged; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000188:drug therapy / Q000235:genetics*; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D066246:ErbB Receptors / Q000235:genetics*; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D009154:Mutation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*
chemical_list: D000970:Antineoplastic Agents; D047428:Protein Kinase Inhibitors; D066246:ErbB Receptors