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Patients with non-muscle invasive bladder cancer (NMIBC) are followed by frequent cystoscopies.Innovative approaches partly replacing cystoscopy (uncomfortable, expensive, low sensitive procedure) aredemanded. The current study aims to establish a fast, reliable, non-invasive, and inexpensive procedure forNMIBC patient surveillance. Liquid biopsy is a reliable source of biomarkers for cancer patient monitoring.Urine is the most suitable biological liquid to search for bladder cancer biomarkers. Cell-free DNA in urinerepresents tumor-related mutations for several cancers, including the bladder. We investigated mutations inFGFR3, TERT promoter, and STAG2 as markers for diagnostics and follow-up in NMIBC. Digital PCRwas used to detect mutations in urine-derived cell-free DNA. The sensitivity and specificity of the markersin relation to clinical outcomes served as criteria of the assay efficiency. The sensitivity with a single marker(TERT) reached 87%, with a specificity of 77%. Combining two biomarkers (TERT+FGFR3) increased thespecificity of the assay to 100% with a sensitivity of 72%. Different mutational status of STAG2 can indicateNMIBC presence or recurrence. Therefore, applying the suggested combination of biomarkers with simpledetection procedures to larger patient cohorts will allow developing procedures for BC detection andsurveillance with optimal sensitivity and specificity. Based on the results of this proof-in-concept study, weconclude that this simple, fast and inexpensive assay can add diagnostic and prognostic value tocystoscopy/cytology analysis of NMIBC patients.

Liquid Biopsy Analysis of FGFR3, TERT Promoter and STAG2 Hotspot Mutations for Disease Surveillance in Bladder Cancer