title: Development and Validation of Urine-based Peptide Biomarker Panels for Detecting Bladder Cancer in a Multi-center Study.
abstract: OBJECTIVE
  Urothelial bladder cancer presents high recurrence rates, mandating continuous monitoring via invasive cystoscopy. The development of noninvasive tests for disease diagnosis and surveillance remains an unmet clinical need. In this study, validation of two urine-based biomarker panels for detecting primary and recurrent urothelial bladder cancer was conducted.
  
  
  METHODS
  Two studies (total n = 1,357) were performed for detecting primary (n = 721) and relapsed urothelial bladder cancer (n = 636). Cystoscopy was applied for detecting urothelial bladder cancer, while patients negative for recurrence had follow-up for at least one year to exclude presence of an undetected tumor at the time of sampling. Capillary electrophoresis coupled to mass spectrometry (CE-MS) was employed for the identification of urinary peptide biomarkers. The candidate urine-based peptide biomarker panels were derived from nested cross-sectional studies in primary (n = 451) and recurrent (n = 425) urothelial bladder cancer.
  
  
  RESULTS
  Two biomarker panels were developed on the basis of 116 and 106 peptide biomarkers using support vector machine algorithms. Validation of the urine-based biomarker panels in independent validation sets, resulted in AUC values of 0.87 and 0.75 for detecting primary (n = 270) and recurrent urothelial bladder cancer (n = 211), respectively. At the optimal threshold, the classifier for detecting primary urothelial bladder cancer exhibited 91% sensitivity and 68% specificity, while the classifier for recurrence demonstrated 87% sensitivity and 51% specificity. Particularly for patients undergoing surveillance, improved performance was achieved when combining the urine-based panel with cytology (AUC = 0.87).
  
  
  CONCLUSIONS
  The developed urine-based peptide biomarker panel for detecting primary urothelial bladder cancer exhibits good performance. Combination of the urine-based panel and cytology resulted in improved performance for detecting disease recurrence. Clin Cancer Res; 22(16); 4077-86. ©2016 AACR.
authors.affiliation: Mosaiques diagnostics GmbH, Hannover, Germany. Biotechnology Division, Biomedical Research Foundation Academy of Athens, Athens, Greece. frantzi@mosaiques-diagnostics.com.
authors.forename: Maria
authors.initials: M
authors.lastname: Frantzi
authors.affiliation: Department of Pathology, Erasmus MC Cancer Institute, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Kim E
authors.initials: KE
authors.lastname: van Kessel
authors.affiliation: Department of Pathology, Erasmus MC Cancer Institute, Erasmus Medical Center, Rotterdam, the Netherlands.
authors.forename: Ellen C
authors.initials: EC
authors.lastname: Zwarthoff
authors.affiliation: Genetic and Molecular Epidemiology Group, Spanish National Cancer Research Center (CNIO), Madrid, Spain.
authors.forename: Mirari
authors.initials: M
authors.lastname: Marquez
authors.affiliation: Genetic and Molecular Epidemiology Group, Spanish National Cancer Research Center (CNIO), Madrid, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Rava
authors.affiliation: Genetic and Molecular Epidemiology Group, Spanish National Cancer Research Center (CNIO), Madrid, Spain.
authors.forename: Núria
authors.initials: N
authors.lastname: Malats
authors.affiliation: Department of Urology, University of Lübeck, Lübeck, Germany.
authors.forename: Axel S
authors.initials: AS
authors.lastname: Merseburger
authors.affiliation: Department of Urology, Laikon Hospital, Medical School of Athens, Athens, Greece.
authors.forename: Ioannis
authors.initials: I
authors.lastname: Katafigiotis
authors.affiliation: Department of Urology, Laikon Hospital, Medical School of Athens, Athens, Greece.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Stravodimos
authors.affiliation: Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: William
authors.initials: W
authors.lastname: Mullen
authors.affiliation: Biotechnology Division, Biomedical Research Foundation Academy of Athens, Athens, Greece.
authors.forename: Jerome
authors.initials: J
authors.lastname: Zoidakis
authors.affiliation: Biotechnology Division, Biomedical Research Foundation Academy of Athens, Athens, Greece.
authors.forename: Manousos
authors.initials: M
authors.lastname: Makridakis
authors.affiliation: Mosaiques diagnostics GmbH, Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Pejchinovski
authors.affiliation: Biotechnology Division, Biomedical Research Foundation Academy of Athens, Athens, Greece.
authors.forename: Elena
authors.initials: E
authors.lastname: Critselis
authors.affiliation: Institute of Clinical Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Ralph
authors.initials: R
authors.lastname: Lichtinghagen
authors.affiliation: Institute of Clinical Chemistry, Hannover Medical School, Hannover, Germany.
authors.forename: Korbinian
authors.initials: K
authors.lastname: Brand
authors.affiliation: Mosaiques diagnostics GmbH, Hannover, Germany.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Dakna
authors.affiliation: Laboratory of Biology, Department of Basic Medical Sciences, University of Athens School of Medicine, Athens, Greece.
authors.forename: Maria G
authors.initials: MG
authors.lastname: Roubelakis
authors.affiliation: University of Colorado, Department of Surgery and Pharmacology, Aurora, Colorado. University of Colorado Comprehensive Cancer Center, Aurora, Colorado.
authors.forename: Dan
authors.initials: D
authors.lastname: Theodorescu
authors.affiliation: Biotechnology Division, Biomedical Research Foundation Academy of Athens, Athens, Greece.
authors.forename: Antonia
authors.initials: A
authors.lastname: Vlahou
authors.affiliation: Mosaiques diagnostics GmbH, Hannover, Germany. Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Harald
authors.initials: H
authors.lastname: Mischak
authors.affiliation: Laboratory of Biology, Department of Basic Medical Sciences, University of Athens School of Medicine, Athens, Greece. Laboratory of Cell and Gene Therapy, Biomedical Research Foundation of the Academy of Athens, Athens, Greece.
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Anagnou
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor; D003430:Cross-Sectional Studies; D003558:Cystoscopy; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D010455:Peptides / Q000652:urine*; D012372:ROC Curve; D012008:Recurrence; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis* / Q000652:urine*
chemical_list: D014408:Biomarkers, Tumor; D010455:Peptides