• PCa vs. HC: NGS identified the top 10 PCa differentially expressed (Log2 fold change > 2) miRNAs in urinary EVs. Higher levels in PCa: miR-10a-5p, miR-204-5p, miR-30a-3p; lower levels in PCa: miR-375, miR-21-5p, miR-141-3p, Let-7C-5p, miR-26b-5p, miR-101-3p, Let-7b-5p.• The diagnostic performance of 3 isomiR combination of miR-21, miR-375, and miR-204 resulted in an AUC of 0.866, compared to a PSA AUC of 0.707 and the corresponding 3 mature microRNAs (miRs) AUC of 0.766. This evidence concerns the gene KLK3 and posterior cortical atrophy.