A multi-gene urinary extracellular vesicle mRNA panel as a non-invasive biomarkers for prostate cancer detection

Prostate cancer (PCa) remains one of the most common malignancies in men older than 50 globally and in India. Although prostate-specific antigen (PSA) remains the standard diagnostic tool, its low specificity leads to overdiagnosis and unnecessary biopsies, highlighting the need for improved non-invasive biomarkers. Urinary extracellular vesicles (UEVs) have emerged as a promising, non-invasive source of molecular biomarkers, particularly RNA, for cancer detection. This study evaluated the diagnostic potential of six PCa associated genes (AGR2, ERG, PTEN, PCA3, AMACR, SPDEF) in UEVs.UEVs were isolated from 15 healthy individuals and 25 PCa patients via polyethylene glycol (PEG) precipitation and characterized by NTA, TEM and western blotting. Gene expression was quantified using qRT-PCR, and diagnostic performance was assessed using logistic regression models and receiver operating characteristic (ROC) analysis. Our results showed that all six genes (AGR2, ERG, PTEN, PCA3, AMACR and SPDEF) were significantly upregulated in PCa patients (p < 0.05) compared to healthy individuals. ROC analyses of these genes yielded area under the curve (AUC) of 0.821, 0.754, 0.746,0.746, 0.730, 0.722 respectively. A combined six-gene logistic model achieved an AUC of 0.96, while a simplified three-gene model1 (AGR2, SPDEF, PCA3) emerged as the most parsimonious model with an AUC of 0.954.Gene expression did not correlate with Gleason score or tumor stage, whereas PSA correlated with higher Gleason score (p = 0.01). These findings highlight the potential of UEV-derived multigene mRNA panels to serve as non-invasive biomarkers for PCa detection.