title: Mutational analysis of the tyrosine kinome in serous and clear cell endometrial cancer uncovers rare somatic mutations in TNK2 and DDR1.
abstract: BACKGROUND
  Endometrial cancer (EC) is the 8th leading cause of cancer death amongst American women. Most ECs are endometrioid, serous, or clear cell carcinomas, or an admixture of histologies. Serous and clear ECs are clinically aggressive tumors for which alternative therapeutic approaches are needed. The purpose of this study was to search for somatic mutations in the tyrosine kinome of serous and clear cell ECs, because mutated kinases can point to potential therapeutic targets.
  
  
  METHODS
  In a mutation discovery screen, we PCR amplified and Sanger sequenced the exons encoding the catalytic domains of 86 tyrosine kinases from 24 serous, 11 clear cell, and 5 mixed histology ECs. For somatically mutated genes, we next sequenced the remaining coding exons from the 40 discovery screen tumors and sequenced all coding exons from another 72 ECs (10 clear cell, 21 serous, 41 endometrioid). We assessed the copy number of mutated kinases in this cohort of 112 tumors using quantitative real time PCR, and we used immunoblotting to measure expression of these kinases in endometrial cancer cell lines.
  
  
  RESULTS
  Overall, we identified somatic mutations in TNK2 (tyrosine kinase non-receptor, 2) and DDR1 (discoidin domain receptor tyrosine kinase 1) in 5.3% (6 of 112) and 2.7% (3 of 112) of ECs. Copy number gains of TNK2 and DDR1 were identified in another 4.5% and 0.9% of 112 cases respectively. Immunoblotting confirmed TNK2 and DDR1 expression in endometrial cancer cell lines. Three of five missense mutations in TNK2 and one of two missense mutations in DDR1 are predicted to impact protein function by two or more in silico algorithms. The TNK2(P761Rfs*72) frameshift mutation was recurrent in EC, and the DDR1(R570Q) missense mutation was recurrent across tumor types.
  
  
  CONCLUSIONS
  This is the first study to systematically search for mutations in the tyrosine kinome in clear cell endometrial tumors. Our findings indicate that high-frequency somatic mutations in the catalytic domains of the tyrosine kinome are rare in clear cell ECs. We uncovered ten new mutations in TNK2 and DDR1 within serous and endometrioid ECs, thus providing novel insights into the mutation spectrum of each gene in EC.
authors.forename: Meghan L
authors.initials: ML
authors.lastname: Rudd
authors.forename: Hassan
authors.initials: H
authors.lastname: Mohamed
authors.forename: Jessica C
authors.initials: JC
authors.lastname: Price
authors.forename: Andrea J
authors.initials: AJ
authors.lastname: O'Hara
authors.forename: Matthieu
authors.initials: M
authors.lastname: Le Gallo
authors.forename: Mary Ellen
authors.initials: ME
authors.lastname: Urick
authors.forename: Pedro
authors.initials: P
authors.lastname: Cruz
authors.forename: Suiyuan
authors.initials: S
authors.lastname: Zhang
authors.forename: Nancy F
authors.initials: NF
authors.lastname: Hansen
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Godwin
authors.forename: Dennis C
authors.initials: DC
authors.lastname: Sgroi
authors.forename: Tyra G
authors.initials: TG
authors.lastname: Wolfsberg
authors.forename: James C
authors.initials: JC
authors.lastname: Mullikin
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Merino
authors.affiliation: Cancer Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda MD 20892, USA. belldaph@mail.nih.gov.
authors.forename: Daphne W
authors.initials: DW
authors.lastname: Bell
mesh_terms: D018262:Adenocarcinoma, Clear Cell / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D018284:Cystadenocarcinoma, Serous / Q000235:genetics* / Q000473:pathology; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis; D004257:DNA Polymerase II / Q000235:genetics; D000070819:Discoidin Domain Receptor 1; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D053842:Microsatellite Instability; D009154:Mutation; D060787:Neoplasm Grading; D000075223:Poly-ADP-Ribose Binding Proteins; D054730:Protein Interaction Domains and Motifs / Q000235:genetics; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics*
chemical_list: D000075223:Poly-ADP-Ribose Binding Proteins; C092030:DDR1 protein, human; D000070819:Discoidin Domain Receptor 1; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; C403262:TNK2 protein, human; D004257:DNA Polymerase II; C119525:POLE protein, human