A Molecular Urine Assay to Detect Recurrences During Surveillance of High-Risk Non-Muscle Invasive Bladder Cancer

Joep J de Jong; Florus C de Jong; Angelique C J van der Made; Niels J van Casteren; Hossain Roshani; Eric H G M Oomens; Rob C M Pelger; Ewout W Steyerberg; Joost L Boormans; Chris H Bangma; Tahlita C M Zuiverloon; Ellen C Zwarthoff

doi:10.3233/BLC-240017

A Molecular Urine Assay to Detect Recurrences During Surveillance of High-Risk Non-Muscle Invasive Bladder Cancer

Bladder Cancer. 2024 Oct 23;10(3):233-242. doi: 10.3233/BLC-240017. eCollection 2024.

Affiliations

¹ Department of Pathology, Erasmus University Medical Center, Rotterdam, The Netherlands.
² Department of Urology, Erasmus MC Urothelial Cancer Research Group, Rotterdam, The Netherlands.
³ Department of Urology, Franciscus Hospital, Rotterdam, The Netherlands.
⁴ Department of Urology, Haga Teaching Hospital, The Hague, The Netherlands.
⁵ Department of Urology, Amphia Hospital, Breda, The Netherlands.
⁶ Department of Urology, Leiden University Medical Center, Leiden, The Netherlands.
⁷ Department of Public Health, Erasmus University Medical Center, Rotterdam, The Netherlands.

Abstract

Background: High-risk non-muscle invasive bladder cancer (HR-NMIBC) patients require long-term surveillance with cystoscopies, cytology and upper tract imaging. Previously, we developed a genomic urine assay for surveillance of HR-NMIBC patients with high sensitivity and anticipatory value.

Objective: We aimed to validate the performance of the assay in an unselected prospectively collected cohort of HR-NMIBC patients under surveillance.

Methods: We included patients from five centers and collected urine sample pairs (evening and morning urines) prior to cystoscopy. Mutation status (FGFR3/TERT) and methylation status (OTX1) was analyzed on DNA from voided urine specimens. A test was considered positive if≥1 alteration was detected in at least one urine sample. The primary endpoint was tumor recurrence. Sensitivity and specificity were determined. A generalized mixed effects model was used to adjust for within-patient correlation. Cox proportional hazard analyses with time-dependent covariates assessed the anticipatory effect of the urine assay.

Results: In total, 204 patients and 736 sample pairs were collected. Sixty-three recurrences were diagnosed for which we had concomitant assay results. On cross-sectional analyses, the assay detected 75% (95% CI 62.1% -84.7%) of recurrences, with a specificity of 70% (95% CI 66.4% -73.5%). Furthermore, mixed effects model analyses revealed OTX1 (p = 0.005) and TERT (p = 0.004) as significant predictors for disease recurrence. Median follow-up was 25.3 months (IQR 18.6-30.7). Twenty-nine tumors were diagnosed without concomitant urine samples, which included recurrences detected after urine collection ended. Longitudinal analyses showed that a positive urine assay predicted a recurrence over time (HR 3.5, p < 0.001). Furthermore, a recurrence during the study period was also a predictor for developing future recurrences (HR 2.1, p < 0.001).

Conclusions: This study validates the performance of a previously developed urine assay in an unselected cohort of HR-NMIBC patients under surveillance. With a robust sensitivity/specificity and a strong anticipatory effect, this assay proves a useful adjunct ready for evaluation in a future randomized controlled trial.

Keywords: Keywords (MeSH): Bladder cancer; methylation; mutation; prospective; urinary biomarkers.