title: Next-Generation Sequencing of Pulmonary Sarcomatoid Carcinoma Reveals High Frequency of Actionable MET Gene Mutations.
abstract: OBJECTIVE
  To further understand the molecular pathogenesis of pulmonary sarcomatoid carcinoma (PSC) and develop new therapeutic strategies in this treatment-refractory disease.
  
  
  METHODS
  Whole-exome sequencing in a discovery set (n = 10) as well as targeted MET mutation screening in an independent validation set (n = 26) of PSC were performed. Reverse transcriptase polymerase chain reaction and Western blotting were performed to validate MET exon 14 skipping. Functional studies for validation of the oncogenic roles of MET exon 14 skipping were conducted in lung adenosquamous cell line H596 (MET exon 14 skipped and PIK3CA mutated) and gastric adenocarcinoma cell line Hs746T (MET exon 14 skipped). Response to MET inhibitor therapy with crizotinib in a patient with advanced PSC and MET exon 14 skipping was evaluated to assess clinical translatability.
  
  
  RESULTS
  In addition to confirming mutations in known cancer-associated genes (TP53, KRAS, PIK3CA, MET, NOTCH, STK11, and RB1), several novel mutations in additional genes, including RASA1, CDH4, CDH7, LAMB4, SCAF1, and LMTK2, were identified and validated. MET mutations leading to exon 14 skipping were identified in eight (22%) of 36 patient cases; one of these tumors also harbored a concurrent PIK3CA mutation. Short interfering RNA silencing of MET and MET inhibition with crizotinib showed marked effects on cell viability and decrease in downstream AKT and mitogen-activated protein kinase activation in Hs746T and H596 cells. Concurrent PIK3CA mutation required addition of a second agent for successful pathway suppression and cell viability effect. Dramatic response to crizotinib was noted in a patient with advanced chemotherapy-refractory PSC carrying a MET exon 14 skipping mutation.
  
  
  CONCLUSIONS
  Mutational events of MET leading to exon 14 skipping are frequent and potentially targetable events in PSC.
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Xuewen
authors.initials: X
authors.lastname: Liu
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Yuxia
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Mark B
authors.initials: MB
authors.lastname: Stoopler
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Yufeng
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Haiying
authors.initials: H
authors.lastname: Cheng
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Jinli
authors.initials: J
authors.lastname: Chen
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Mahesh
authors.initials: M
authors.lastname: Mansukhani
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Sanjay
authors.initials: S
authors.lastname: Koul
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA. bahalmos@montefiore.org.
authors.forename: Balazs
authors.initials: B
authors.lastname: Halmos
authors.affiliation: Xuewen Liu, Mark B. Stoopler, Yufeng Shen, Jinli Chen, Mahesh Mansukhani, Sanjay Koul, Balazs Halmos, and Alain C. Borczuk, Columbia University Medical Center; Haiying Cheng, Montefiore Medical Center/Albert Einstein College of Medicine, New York, NY; Xuewen Liu, Sun Yat-sen University Cancer Center, Guangzhou, People's Republic of China; and Yuxia Jia, Penn State Milton S. Hershey Medical Center, Penn State College of Medicine, Hershey, PA.
authors.forename: Alain C
authors.initials: AC
authors.lastname: Borczuk
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D048708:Cell Growth Processes / Q000502:physiology; D058534:Class I Phosphatidylinositol 3-Kinases; D015331:Cohort Studies; D000077547:Crizotinib; D004252:DNA Mutational Analysis / Q000379:methods; D004273:DNA, Neoplasm / Q000235:genetics; D005091:Exons; D005260:Female; D055785:Gene Knockdown Techniques; D006801:Humans; D007527:Isoenzymes; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics; D017354:Point Mutation; D047428:Protein Kinase Inhibitors / Q000627:therapeutic use; D019859:Proto-Oncogene Proteins c-met / Q000037:antagonists & inhibitors / Q000235:genetics*; D011720:Pyrazoles / Q000627:therapeutic use; D011725:Pyridines / Q000627:therapeutic use; D012509:Sarcoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology
chemical_list: D004273:DNA, Neoplasm; D007527:Isoenzymes; D047428:Protein Kinase Inhibitors; D011720:Pyrazoles; D011725:Pyridines; D000077547:Crizotinib; D019869:Phosphatidylinositol 3-Kinases; D058534:Class I Phosphatidylinositol 3-Kinases; C484760:PIK3CA protein, human; C491743:MET protein, human; D019859:Proto-Oncogene Proteins c-met