title: KIF5B-ALK, a novel fusion oncokinase identified by an immunohistochemistry-based diagnostic system for ALK-positive lung cancer.
abstract: OBJECTIVE
  EML4-ALK is a transforming fusion tyrosine kinase, several isoforms of which have been identified in lung cancer. Immunohistochemical detection of EML4-ALK has proved difficult, however, likely as a result of low transcriptional activity conferred by the promoter-enhancer region of EML4. The sensitivity of EML4-ALK detection by immunohistochemistry should be increased adequately.
  
  
  METHODS
  We developed an intercalated antibody-enhanced polymer (iAEP) method that incorporates an intercalating antibody between the primary antibody to ALK and the dextran polymer-based detection reagents.
  
  
  RESULTS
  Our iAEP method discriminated between tumors positive or negative for EML4-ALK in a test set of specimens. Four tumors were also found to be positive for ALK in an archive of lung adenocarcinoma (n = 130) and another 4 among fresh cases analyzed in a diagnostic laboratory. These 8 tumors were found to include 1 with EML4-ALK variant 1, 1 with variant 2, 3 with variant 3, and 2 with previously unidentified variants (designated variants 6 and 7). Inverse reverse transcription-PCR analysis revealed that the remaining tumor harbored a novel fusion in which intron 24 of KIF5B was ligated to intron 19 of ALK. Multiplex reverse transcription-PCR analysis of additional archival tumor specimens identified another case of lung adenocarcinoma positive for KIF5B-ALK.
  
  
  CONCLUSIONS
  The iAEP method should prove suitable for immunohistochemical screening of tumors positive for ALK or ALK fusion proteins among pathologic archives. Coupling of PCR-based detection to the iAEP method should further facilitate the rapid identification of novel ALK fusion genes such as KIF5B-ALK.
authors.affiliation: The Cancer Institute, Japanese Foundation for Cancer Research, Japan. kentakeuchi-tky@umin.net
authors.forename: Kengo
authors.initials: K
authors.lastname: Takeuchi
authors.forename: Young Lim
authors.initials: YL
authors.lastname: Choi
authors.forename: Yuki
authors.initials: Y
authors.lastname: Togashi
authors.forename: Manabu
authors.initials: M
authors.lastname: Soda
authors.forename: Satoko
authors.initials: S
authors.lastname: Hatano
authors.forename: Kentaro
authors.initials: K
authors.lastname: Inamura
authors.forename: Shuji
authors.initials: S
authors.lastname: Takada
authors.forename: Toshihide
authors.initials: T
authors.lastname: Ueno
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yamashita
authors.forename: Yukitoshi
authors.initials: Y
authors.lastname: Satoh
authors.forename: Sakae
authors.initials: S
authors.lastname: Okumura
authors.forename: Ken
authors.initials: K
authors.lastname: Nakagawa
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Ishikawa
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Mano
mesh_terms: D016475:3T3 Cells; D000230:Adenocarcinoma / Q000201:enzymology*; D000077548:Anaplastic Lymphoma Kinase; D000818:Animals; D000911:Antibodies, Monoclonal; D002289:Carcinoma, Non-Small-Cell Lung / Q000201:enzymology*; D002471:Cell Transformation, Neoplastic; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D014644:Genetic Variation; D006801:Humans; D007124:Immunoenzyme Techniques; D016547:Kinesins / Q000235:genetics / Q000378:metabolism*; D008175:Lung Neoplasms / Q000201:enzymology*; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D015514:Oncogene Proteins, Fusion / Q000235:genetics / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry; D011379:Prognosis; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020794:Receptor Protein-Tyrosine Kinases; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000911:Antibodies, Monoclonal; C522504:EML4-ALK fusion protein, human; C488101:KIF5B protein, human; D015514:Oncogene Proteins, Fusion; D011108:Polymers; D012333:RNA, Messenger; C000626173:ALK protein, human; C000626174:Alk protein, mouse; D000077548:Anaplastic Lymphoma Kinase; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D016547:Kinesins