Fibroblast Growth Factor Receptor 3 Mutation Analysis on Voided Urine for Surveillance of Patients with Low-Grade Non-Muscle–Invasive Bladder Cancer

Abstract

Purpose: Mutations in the fibroblast growth factor receptor 3 (FGFR3) have been found in 70% of the low-grade non-muscle-invasive bladder cancer (NMI-BC) tumors. We aim to determine the potential of FGFR3 mutation analysis on voided urine to detect recurrences during surveillance of patients with lowgrade NMI-BC.Experimental Design: FGFR3 mutation status of the study inclusion tumor was determined from 200 low-grade NMI-BC patients. Patients with an FGFR3-mutant inclusion tumor were selected for analysis and monitored by cystoscopy, and voided urine samples were collected. FGFR3 mutation analysis was done on 463 prospectively collected urines. Sensitivity and predictive value of the assay were determined for detection of concomitant recurrences. Longitudinal and Cox time-to-event analyses were done to determine the predictive value for detection of future recurrences.Results: Median follow-up was 3.5 years. The sensitivity of the assay for detection of concomitant recurrences was 26 of 45 (58%). Of the 105 positive urine samples, 85 (81%) were associated with a concomitant or a future recurrence. An FGFR3-positive urine was associated with a 3.8-fold (P < 0.0001) higher risk of having a recurrence in the Cox analysis. In contrast, only 41 of 358 (11%) FGFR3-negative urine samples were associated with a recurrence. Positive predictive value increased from 25% to 90% in patients having consecutive FGFR3-positive urine tests.Conclusions: FGFR3 mutation analysis on voided urine is a simple and noninvasive diagnostic method for detection of recurrences during surveillance of patients presenting with a low-grade FGFR3-mutant NMI-BC tumor. Clin Cancer Res; 16(11); 3011-8. ©2010 AACR.Bladder cancer (BC) is the fifth most common malignancy in the world and can be divided into non-muscleinvasive BC (NMI-BC) and muscle-invasive BC tumors ref. 1). Almost 80% of the NMI-BC patients will present with a G1 or G2, low-grade NMI tumor, making this a considerable burden on the urological practice. The prognosis of low-grade NMI-BC patients is good with a 5-year survival of 80% to 90% (2, 3). NMI bladder tumors are treated by transurethral resection, but 70% of the patients will develop at least one recurrence within 5 years and 10% to 20% will progress to MI disease. Currently, the surveillance protocol consists of cystoscopy every 3 to 6 months for the first 2 years, followed by less frequent observations when a patient stays recurrence-free (4). The frequent transurethral resections and follow-up cystoscopies make BC from diagnosis to death the most costly cancer in the world (5-8). The life-long follow-up of patients by invasive cystoscopy and associated high costs emphasize the need for a urinary biomarker, which can be used in a noninvasive assay for the detection of recurrences and to stratify patients with a high-risk profile, hereby creating the possibility to reduce the number of cystoscopies and thus improving quality of life.About 70% of low-grade NMI tumors have a mutation in the fibroblast growth factor rece...

A Panel of Three Markers Hyper- and Hypomethylated in Urine Sediments Accurately Predicts Bladder Cancer Recurrence

“…More recently, it has been proposed that longitudinal collection and testing of urine sediments may help assess the prognostic and recurrence predictive value of markers (43,44). Several studies undertook this approach by using DNA methylation analysis, microsatellite markers, and a FGFR3 mutation assay (45,46). Although these markers were highly sensitive, they displayed low specificity in some cases comparable with that of cytology or a high rate of false-positive results (47,48).…”

Roles of Fibroblast Growth Factor Receptors in Carcinogenesis

“…The high recurrence rate (60-80%) of noninvasive tumors requires long-term, expensive patient monitoring. Recent data suggest that detection of FGFR3 mutations in urine from patients with FGFR3 mutations in the primary tumor indicates recurrence (101,102). Thus, identification of FGFR3 mutations is not only a potential biomarker for bladder cancer diagnosis and prognosis but could also indicate tumor recurrence.…”

A 3-Plex Methylation Assay Combined with the FGFR3 Mutation Assay Sensitively Detects Recurrent Bladder Cancer in Voided Urine

“…We further developed a multiplex BS-SNaPshot assay aimed at detecting the methylation status of the 3 most interesting CGIs (OTX1, ONECUT2, and OSR1) in a single PCR and a single nucleotide primer extension reaction. FGFR3 mutation analysis was conducted as described previously (31). Concentrations of all primers and probes are given in Supplementary Table S1.…”