title: Telomerase reverse transcriptase promoter mutations in bladder cancer: high frequency across stages, detection in urine, and lack of association with outcome.
abstract: BACKGROUND
  Hotspot mutations in the promoter of the gene coding for telomerase reverse transcriptase (TERT) have been described and proposed to activate gene expression.
  
  
  OBJECTIVE
  To investigate TERT mutation frequency, spectrum, association with expression and clinical outcome, and potential for detection of recurrences in urine in patients with urothelial bladder cancer (UBC).
  
  
  METHODS
  A set of 111 UBCs of different stages was used to assess TERT promoter mutations by Sanger sequencing and TERT messenger RNA (mRNA) expression by reverse transcription-quantitative polymerase chain reaction. The two most frequent mutations were investigated, using a SNaPshot assay, in an independent set of 184 non-muscle-invasive and 173 muscle-invasive UBC (median follow-up: 53 mo and 21 mo, respectively). Voided urine from patients with suspicion of incident UBC (n=174), or under surveillance after diagnosis of non-muscle-invasive UBC (n=194), was tested using a SNaPshot assay.
  
  
  METHODS
  Association of mutation status with age, sex, tobacco, stage, grade, fibroblast growth factor receptor 3 (FGFR3) mutation, progression-free survival, disease-specific survival, and overall survival.
  
  
  CONCLUSIONS
  In the two series, 78 of 111 (70%) and 283 of 357 (79%) tumors harbored TERT mutations, C228T being the most frequent substitution (83% for both series). TERT mutations were not associated with clinical or pathologic parameters, but were more frequent among FGFR3 mutant tumors (p=0.0002). There was no association between TERT mutations and mRNA expression (p=0.3). Mutations were not associated with clinical outcome. In urine, TERT mutations had 90% specificity in subjects with hematuria but no bladder tumor, and 73% in recurrence-free UBC patients. The sensitivity was 62% in incident and 42% in recurrent UBC. A limitation of the study is its retrospective nature.
  
  
  CONCLUSIONS
  Somatic TERT promoter mutations are an early, highly prevalent genetic event in UBC and are not associated with TERT mRNA levels or disease outcomes. A SNaPshot assay in urine may help to detect UBC recurrences.
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain; Université Paris-Est Créteil, Institut Mondor de Recherche Biomédicale, Créteil, France.
authors.forename: Yves
authors.initials: Y
authors.lastname: Allory
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Willemien
authors.initials: W
authors.lastname: Beukers
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Sagrera
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Flández
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Marqués
authors.affiliation: Genetic and Molecular Epidemiology Group, Human Cancer Genetics Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Mirari
authors.initials: M
authors.lastname: Márquez
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Kirstin A
authors.initials: KA
authors.lastname: van der Keur
authors.affiliation: Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Lars
authors.initials: L
authors.lastname: Dyrskjot
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Irene
authors.initials: I
authors.lastname: Lurkin
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Marcel
authors.initials: M
authors.lastname: Vermeij
authors.affiliation: Medical Oncology Department, Ramón y Cajal University Hospital, Madrid, Spain.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Carrato
authors.affiliation: Department of Pathology, Hospital del Mar-Parc de Salut Mar, Barcelona, Spain; Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain.
authors.forename: Josep
authors.initials: J
authors.lastname: Lloreta
authors.affiliation: Urology Service, Hospital del Mar-Parc de Salut Mar, Barcelona, Spain.
authors.forename: José A
authors.initials: JA
authors.lastname: Lorente
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Enrique
authors.initials: E
authors.lastname: Carrillo-de Santa Pau
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Roy G
authors.initials: RG
authors.lastname: Masius
authors.affiliation: Centre de Recerca d'Epidemiologia Ambiental, Barcelona, Spain; IMIM-Institut de Recerca Hospital del Mar, Barcelona, Spain; CIBER Epidemiología y Salud Pública (CIBERESP), Barcelona, Spain; National School of Public Health, Athens, Greece.
authors.forename: Manolis
authors.initials: M
authors.lastname: Kogevinas
authors.affiliation: Department of Public Health, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Ewout W
authors.initials: EW
authors.lastname: Steyerberg
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Angela A G
authors.initials: AA
authors.lastname: van Tilborg
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands; Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Cheno
authors.initials: C
authors.lastname: Abas
authors.affiliation: Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark.
authors.forename: Torben F
authors.initials: TF
authors.lastname: Orntoft
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Tahlita C M
authors.initials: TC
authors.lastname: Zuiverloon
authors.affiliation: Genetic and Molecular Epidemiology Group, Human Cancer Genetics Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain.
authors.forename: Núria
authors.initials: N
authors.lastname: Malats
authors.affiliation: Department of Pathology, Erasmus MC, Rotterdam, The Netherlands.
authors.forename: Ellen C
authors.initials: EC
authors.lastname: Zwarthoff
authors.affiliation: Epithelial Carcinogenesis Group, Molecular Pathology Program, CNIO (Spanish National Cancer Research Center), Madrid, Spain; Departament de Ciències Experimentals i de la Salut, Universitat Pompeu Fabra, Barcelona, Spain. Electronic address: preal@cnio.es.
authors.forename: Francisco X
authors.initials: FX
authors.lastname: Real
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000652:urine; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D018450:Disease Progression; D018572:Disease-Free Survival; D005260:Female; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D008297:Male; D009154:Mutation; D060787:Neoplasm Grading; D009364:Neoplasm Recurrence, Local; D009367:Neoplasm Staging; D009426:Netherlands; D010641:Phenotype; D011237:Predictive Value of Tests; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000652:urine; D012189:Retrospective Studies; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012307:Risk Factors; D013030:Spain; D019098:Telomerase / Q000235:genetics* / Q000652:urine; D013997:Time Factors; D001749:Urinary Bladder Neoplasms / Q000201:enzymology* / Q000235:genetics* / Q000401:mortality / Q000473:pathology / Q000628:therapy / Q000652:urine
chemical_list: D014408:Biomarkers, Tumor; D012333:RNA, Messenger; C509186:TERT protein, human; D019098:Telomerase
keywords: Bladder cancer; Clinical end point; Somatic mutations; TERT gene; Urine-based diagnosis