A Novel Methylation Marker NRN1 plus TERT and FGFR3 Mutation Using Urine Sediment Enables the Detection of Urothelial Bladder Carcinoma

Abstract

Background: Aberrant DNA methylation is an early event during tumorigenesis. In the present study, we aimed to construct a methylation diagnostic tool using urine sediment for the detection of urothelial bladder carcinoma, and improved the diagnostic performance of the model by incorporating single-nucleotide polymorphism (SNP) sites. Methods: A three-stage analysis was carried out to construct the model and evaluate the diagnostic performance. In stage I, two small cohorts from Xiangya hospital were recruited to validate and identify the detailed regions of collected methylation biomarkers. In stage II, proof-of-concept study cohorts from the Hunan multicenter were recruited to construct a diagnostic tool. In stage III, a blinded cohort comprising suspicious UBC patients was recruited from Beijing single center to further test the robustness of the model. Results: In stage I, single NRN1 exhibited the highest AUC compared with six other biomarkers and the Random Forest model. At the best cutoff value of 5.16, a single NRN1 biomarker gave a diagnosis with a sensitivity of 0.93 and a specificity of 0.97. In stage II, the Random Forest algorithm was applied to construct a diagnostic tool, consisting of NRN1, TERT C228T and FGFR3 p.S249C. The tool exhibited AUC values of 0.953, 0.946 and 0.951 in training, test and all cohorts. At the best cutoff value, the model resulted in a sensitivity of 0.871 and a specificity of 0.947. In stage III, the diagnostic tool achieved a good discrimination in the external validation cohort, with an overall AUC of 0.935, sensitivity of 0.864 and specificity of 0.895. Additionally, the model exhibited a superior sensitivity and comparable specificity compared with conventional cytology and FISH. Conclusions: The diagnostic tool exhibited a highly specific and robust performance. It may be used as a replaceable approach for the detection of UBC.

Urinary DNA methylation‐based risk stratification model to triage patients for repeat transurethral resection of bladder tumours

“…Wei Ouyang 1 Ran Xu 2 Hanyu Yao 1 Shusuan Jiang 3 Qiang Lu 4 Cai Lv 5 Pei Li 6 Genming Xu 6 Jianye Liu 1 Long Wang…”

“…Our previous study showed that the methylation status of NRN1 exhibits remarkable performance in detecting urothelial carcinoma and can be used for its diagnosis. 2 , 3 In this prospective, multicenter study, we developed a novel risk‐stratification model based on urine biopsy to identify residual tumours after initial TURBT and to avoid unnecessary re‐TURBT procedures.…”

“…The methods of sample collection, DNA extraction and DNA methylation detection are described in our previously published study. 3 , 4 The clinical characteristics of enrolled cases are shown in Table S1 . The residual rate after initial TURBT was 30.77% (24/78).…”

Advancing Genomics in Urologic Tumors: Navigating Precision Therapeutic Pathways

“…As research in this area continues to progress, the integration of methylation markers and SNP sites into diagnostic protocols is poised to become a cornerstone of UBC management, offering renewed hope and improved care for patients affected by this challenging malignancy [34]. As the global population ages and environmental exposures continue to evolve, understanding the epidemiology, risk factors, and early detection strategies for bladder cancer becomes increasingly imperative [35].…”

Urine-Based Biomarker Test Uromonitor® in the Detection and Disease Monitoring of Non-Muscle-Invasive Bladder Cancer—A Systematic Review and Meta-Analysis of Diagnostic Test Performance

“…Among urine-based diagnostic tests (UBDT), only urinary cytology is widely established in clinical practice, although it exhibits limited sensitivity in low-grade (LG) UCB and heavily relies on the expertise of the examiner [2]. Over the past two decades, several UBDTs have been developed in an effort to avoid unnecessary cystoscopies in the oncological follow-up of NMIBC patients without compromising patient safety [3][4][5][6][7][8][9][10][11][12][13][14][15][16][17]. Cystoscopic follow-up, in particular, is associated with significant discomfort for NMIBC patients and significantly contributes to the high long-term costs of this disease [18].…”