Validation of a DNA methylation-mutation urine assay to select patients with hematuria for cystoscopy

Only 3-28% of patients referred to the urology clinic for hematuria are diagnosed with bladder cancer (BC). Cystoscopies lead to high diagnostic costs and patient burden. Therefore, to improve selection of patients for cystoscopy, reduce costs and over-testing we aimed to validate a recently developed diagnostic urine assay MATERIALS AND METHODS: We included 200 patients from three European countries undergoing cystoscopy for hematuria (n=97 BC, n=103 non-malignant). Voided urine samples were collected prior to cystoscopy. DNA was extracted and analyzed for mutations in FGFR3, TERT and HRAS and methylation of OTX1, ONECUT2 and TWIST1. Logistic regression was used to analyze the association between predictor variables and BC.

Combining the methylation and mutation markers with age led to an AUC of 0.96 (95% CI 0.92-0.99, sensitivity 93% and specificity 86%) and an optimism-corrected AUC of 0.95. The AUC was higher for ≥T1 tumors (0.99) compared to Ta tumors (0.93). The AUC was also higher in high grade tumors (1.00) compared to low grade tumors (0.93). The overall negative predictive value was 99%, based on 5-10% prevalence of BC in patients with hematuria. This would lead to 77% reduction in diagnostic cystoscopies.

Analyzing hematuria patients for their risk of BC using novel molecular markers may lead to a reduction of diagnostic cystoscopies. Combining methylation analysis (OTX1, ONECUT2 and TWIST1) with mutation analysis (FGFR3, TERT and HRAS) and age resulted in a validated accurate prediction model.

The Journal of urology. 2016 Oct 13 [Epub ahead of print]

Kim E M van Kessel, Willemien Beukers, Irene Lurkin, Angelique Ziel-van der Made, Kirstin A van der Keur, Joost L Boormans, Lars Dyrskjøt, Mirari Márquez, Torben F Ørntoft, Francisco X Real, Ulrika Segersten, Núria Malats, Per-Uno Malmström, Wim Van Criekinge, Ellen C Zwarthoff

Department of Pathology, Erasmus Medical Center, Rotterdam, the Netherlands., Department of Urology, Erasmus Medical Center, Rotterdam, the Netherlands., Department of Molecular Medicine, Aarhus University Hospital, Aarhus, Denmark., Genetic and Molecular Epidemiology Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain., Epithelial Carcinogenesis Group, Spanish National Cancer Research Centre (CNIO), Madrid, Spain; Department of Experimental and Health Sciences, Universitat Pompeu Fabra, Barcelona, Spain., Department of Surgical Sciences, Uppsala University, Uppsala, Sweden., MDxHealth, Inc., Irvine, CA, USA; Laboratory of Bioinformatics and Computational Genomics, Ghent University, Belgium., Department of Pathology, Erasmus Medical Center, Rotterdam, the Netherlands. Electronic address: .