Development and validation of a urine-based DNA methylation assay for early endometrial cancer detection: multicenter study

Endometrial cancer (EC) incidence is rising globally, yet diagnosis still depends of invasive procedures, which limit accessibility and delay early detection. A noninvasive diagnostic assay is urgently needed to enable early EC detection. A diagnostic study across six centers in China integrated whole-epigenome methylation profiling of 1211 datasets followed by targeted bisulfite sequencing of 367 samples to identify EC-specific urine DNA methylation biomarkers. A qPCR-based urinary methylation assay (‘Urine-qEC’) targeting three genes (VWC2, ZNF132, and ZSCAN12) was subsequently developed and validated in a cross-sectional cohort (n=587). Urine hypermethylation signatures exhibited concordance with EC tumors and significantly diminished after tumor resection. The Urine-qEC assay achieved an AUC of 0.976 (sensitivity 89.4%, specificity 97.0%) in the training cohort and 0.938 (sensitivity 85.5%, specificity 91.1%) in the multicenter validation. Sensitivity remained high across clinical stage, while specificity was consistent across control subgroups, reaching 93.0% against symptomatic controls with abnormal uterine bleeding and 91.0% against benign controls. The authors conclude that the Urine-qEC assay demonstrated the technical feasibility of a urine-based methylation approach for noninvasive endometrial cancer detection.