title: Functional kinomics identifies candidate therapeutic targets in head and neck cancer.
abstract: OBJECTIVE
  To identify novel therapeutic drug targets for p53-mutant head and neck squamous cell carcinoma (HNSCC).
  
  
  METHODS
  RNAi kinome viability screens were performed on HNSCC cells, including autologous pairs from primary tumor and recurrent/metastatic lesions, and in parallel on murine squamous cell carcinoma (MSCC) cells derived from tumors of inbred mice bearing germline mutations in Trp53, and p53 regulatory genes: Atm, Prkdc, and p19(Arf). Cross-species analysis of cell lines stratified by p53 mutational status and metastatic phenotype was used to select 38 kinase targets. Both primary and secondary RNAi validation assays were performed on additional HNSCC cell lines to credential these kinase targets using multiple phenotypic endpoints. Kinase targets were also examined via chemical inhibition using a panel of kinase inhibitors. A preclinical study was conducted on the WEE1 kinase inhibitor, MK-1775.
  
  
  RESULTS
  Our functional kinomics approach identified novel survival kinases in HNSCC involved in G2-M cell-cycle checkpoint, SFK, PI3K, and FAK pathways. RNAi-mediated knockdown and chemical inhibition of the WEE1 kinase with a specific inhibitor, MK-1775, had a significant effect on both viability and apoptosis. Sensitivity to the MK-1775 kinase inhibitor is in part determined by p53 mutational status, and due to unscheduled mitotic entry. MK-1775 displays single-agent activity and potentiates the efficacy of cisplatin in a p53-mutant HNSCC xenograft model.
  
  
  CONCLUSIONS
  WEE1 kinase is a potential therapeutic drug target for HNSCC. This study supports the application of a functional kinomics strategy to identify novel therapeutic targets for cancer.
authors.affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington.
authors.forename: Russell
authors.initials: R
authors.lastname: Moser
authors.affiliation: Department of Otolaryngology: Head and Neck Surgery, University of Washington Medical Center, Seattle, Washington. Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington.
authors.forename: Chang
authors.initials: C
authors.lastname: Xu
authors.affiliation: Department of Otolaryngology: Head and Neck Surgery, University of Washington Medical Center, Seattle, Washington.
authors.forename: Michael
authors.initials: M
authors.lastname: Kao
authors.affiliation: Quellos High Throughput Facility, Institute for Stem Cell And Regenerative Medicine, University of Washington Medicine Research, Seattle, Washington.
authors.forename: James
authors.initials: J
authors.lastname: Annis
authors.affiliation: Department of Otolaryngology: Head and Neck Surgery, University of Washington Medical Center, Seattle, Washington.
authors.forename: Luisa Angelica
authors.initials: LA
authors.lastname: Lerma
authors.affiliation: Toxicology Program, Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, Washington.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Schaupp
authors.affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington.
authors.forename: Kay E
authors.initials: KE
authors.lastname: Gurley
authors.affiliation: Sage Bionetworks, Seattle, Washington.
authors.forename: In Sock
authors.initials: IS
authors.lastname: Jang
authors.affiliation: Deparment of Surgery, Otolaryngology: Head and Neck Surgery, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Asel
authors.initials: A
authors.lastname: Biktasova
authors.affiliation: Deparment of Surgery, Otolaryngology: Head and Neck Surgery, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Wendell G
authors.initials: WG
authors.lastname: Yarbrough
authors.affiliation: Sage Bionetworks, Seattle, Washington.
authors.forename: Adam A
authors.initials: AA
authors.lastname: Margolin
authors.affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington. Quellos High Throughput Facility, Institute for Stem Cell And Regenerative Medicine, University of Washington Medicine Research, Seattle, Washington.
authors.forename: Carla
authors.initials: C
authors.lastname: Grandori
authors.affiliation: Division of Human Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington. edmendez@u.washington.edu cjkemp@fhcrc.org.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Kemp
authors.affiliation: Department of Otolaryngology: Head and Neck Surgery, University of Washington Medical Center, Seattle, Washington. Division of Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, Washington. Surgery and Perioperative Care Service, VA Puget Sound Health Care System, Seattle, Washington. edmendez@u.washington.edu cjkemp@fhcrc.org.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Méndez
mesh_terms: D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D006258:Head and Neck Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D016159:Tumor Suppressor Protein p53 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D034741:RNA, Small Interfering; D016159:Tumor Suppressor Protein p53; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; C499348:Wee1 protein, mouse