title: Identification of novel mutations in Japanese ovarian clear cell carcinoma patients using optimized targeted NGS for clinical diagnosis.
abstract: OBJECTIVE
  Ovarian clear cell carcinoma (OCCC) is an aggressive ovarian cancer with a higher frequency in Japan and often becomes chemorefractory disease. Reliable genetic diagnosis is essential to affirm the success of precision medicine for OCCC treatment. The aim of this study is, therefore, to identify novel mutations in OCCCs and develop a feasible clinical next generation sequencing (NGS) approach using formalin-fixed paraffin-embedded (FFPE) rather than preferable but not always available fresh frozen (FF) samples.
  
  
  METHODS
  We optimized and evaluated exome analyses of 409 cancer-related genes using FFPE and FF DNA and analyzed NGS data to identify somatic mutations in Japanese OCCCs.
  
  
  RESULTS
  Sufficient and good quality DNAs from FFPE samples were extracted from 18 (FIGO Stage I: 12) out of 29 pairs of matched normal and OCCC for NGS (63%). The fine quality of extracted DNAs depended on the length of storage period (<2years storage). We also identified 45 somatic mutations in 34 genes including unreported variants from those FFPE DNA, in which somatic mutations in the PIK3CA gene was the most common (28%) as previously reported. Seven genes (PIK3CA, ARID1A, CTNNB1, CSMD3, LPHN3, LRP1B, and TP53) were mutated in at least two independent OCCCs. FF samples from 3 out of those 18 OCCCs were available and 13 out of 14 FFPE somatic mutations were confirmed.
  
  
  CONCLUSIONS
  We successfully identified novel genetic alterations in Japanese OCCCs and demonstrated a feasible clinical diagnostic procedure using targeted NGS for OCCC FFPE samples.
authors.affiliation: Department of Molecular Carcinogenesis, Chiba Cancer Center Research Institute, Chiba, Japan; Department of Molecular Biology and Oncology, Graduate School of Medical and Pharmaceutical Sciences, Chiba University, Chiba, Japan.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Maru
authors.affiliation: Department of Gynecology, Chiba Cancer Center, Chiba, Japan.
authors.forename: Naotake
authors.initials: N
authors.lastname: Tanaka
authors.affiliation: Laboratory of Cancer Genomics, Chiba Cancer Center Research Institute, Chiba, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Ohira
authors.affiliation: Division of Surgical Pathology, Chiba Cancer Center, Chiba, Japan.
authors.forename: Makiko
authors.initials: M
authors.lastname: Itami
authors.affiliation: Department of Molecular Carcinogenesis, Chiba Cancer Center Research Institute, Chiba, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Hippo
authors.affiliation: Department of Molecular Biology and Oncology, Graduate School of Medical and Pharmaceutical Sciences, Chiba University, Chiba, Japan; Laboratory of Cancer Genetics, Chiba Cancer Center Research Institute, Chiba, Japan. Electronic address: hnagase@chiba-cc.jp.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Nagase
mesh_terms: D018262:Adenocarcinoma, Clear Cell / Q000235:genetics* / Q000473:pathology; D000328:Adult; D000368:Aged; D005260:Female; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008875:Middle Aged; D009154:Mutation; D010051:Ovarian Neoplasms / Q000235:genetics* / Q000473:pathology; D016612:Paraffin Embedding
keywords: Clinical sequencing; FFPE; Next-generation sequencing; Ovarian clear cell carcinoma; Somatic mutations