Validation of a microRNA profile in urine liquid biopsy with diagnostic and stratification value for bladder cancer classification, available through the open app BladdermiRaCan

• Published in: Experimental hematology & oncology Vol. 14; no. 1; pp. 58 - 6
• Main Authors: Oto, Julia, Herranz, Raquel, Plana, Emma, Pérez-Ardavín, Javier, Hervás, David, Cana, Fernando, Verger, Patricia, Ramos-Soler, David, Martínez-Sarmiento, Manuel, Vera-Donoso, César D., Medina, Pilar
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 11.04.2025; BMC
• Subjects: Analysis; Biomarker; Bladder cancer; BladdermiRaCan; Diagnosis; Genetic aspects; Health aspects; Liquid biopsy; MicroRNA; miR-eCLIP Immunoprecipitation; Urine; Spain; Urine; Biomarker; Prognosis; BladdermiRaCan; Liquid biopsy; microRNA; Diagnosis; Stratification; Bladder cancer; miR-eCLIP Immunoprecipitation
• DOI: 10.1186/s40164-025-00649-0
• ISSN: 2162-3619

We aimed to identify a profile of urine microRNAs (miRNAs) with diagnostic and stratification potential in the whole range of bladder cancer (BC) categories, to avoid current invasive, harmful and expensive procedures. We collected a first morning urine sample from the screening (35 BC patients and 15 age- and gender-matched controls) and validation cohorts (172 BC and 94 controls). In the screening stage we analyzed the expression level of 179 miRNAs by real-time reverse transcription quantitative PCR in urine supernatants. miRNA levels in each sample were normalized by the levels of the previously identified and stably expressed miR-29c-3p. We performed an ordinal regression for each miRNA with False Discovery Rate (FDR) adjustment to identify dysregulated miRNAs, and an ordinal elastic net logistic regression model to identify a miRNA profile for BC diagnosis and stratification with the software R (v3.5.1). Next, we validated the most dysregulated miRNAs, and empirically identified the real miRNA targets in BC cells by miR-eCLIP immunoprecipitation and sequencing. We identified 70 dysregulated miRNAs in BC patients ( p < 0.05 FDR-adjusted). With the expression level of 7 miRNAs in urine (miR-221-3p, miR-93-5p, miR-362-3p, miR-191-5p, miR-200c-3p, miR-192-5p, miR-21-5p) we could stratify BC patients and control subjects. To enable the global use of our model, we developed the free BladdermiRaCan online tool. Furthermore, we identified miR-21-5p, miR-425-5p and miR-99a-5p as follow-up markers for BC relapse, and miR-21-5p and miR-221-3p as markers for metastasis. These miRNAs were also dysregulated in BC tissue sections from a subgroup of patients from which urine samples were studied. In conclusion, we have validated and patented a 7-miRNAs urine profile able to diagnose and stratify BC patients; BladdermiRaCan will enable the global use of our model. The experimentally verified target proteins identified for these miRNAs may unravel novel therapeutic targets.