Department of Urology, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Department of Pathology, Cancer Center Amsterdam, Amsterdam UMC, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Clin Epigenetics. 2022 Feb 5;14(1):19. doi: 10.1186/s13148-022-01240-8.
The development of accurate urinary biomarkers for non-invasive and cost-effective detection of primary and recurrent bladder tumours is recognized as one of the major clinical needs in bladder cancer diagnostics. The purposes of this study were (1) to validate the results of a previous technical comparison by determining the diagnostic performance of nine methylation markers in urine pellet compared to full void urine, and (2) to validate the diagnostic performance of the optimal marker panel GHSR/MAL from a previous exploratory study in a preclinical setting.
Urine samples of 108 patients with bladder cancer and 100 age- and gender-matched controls were prospectively collected for methylation analysis. Urinary methylation levels of the markers FAM19A4, GHSR, MAL, miR-129, miR-935, PHACTR3, PRDM14, SST and ZIC1 were determined with quantitative methylation-specific PCR in urine pellet. Area under the curves (AUCs) were determined for individual markers and the marker panel GHSR/MAL. The diagnostic performance of the marker panel GHSR/MAL was evaluated in the total study population and in different subgroups of patients with bladder cancer using the Chi-square test. The diagnostic accuracy was assessed by leave-one-out cross-validation.
All nine urinary methylation markers (FAM19A4, GHSR, MAL, miR-129, miR-935, PHACTR3, PRDM14, SST and ZIC1) showed significantly higher methylation levels in bladder cancer patients than in controls (p < 0.001). Area under the curves (AUCs) of the nine methylation markers tested in urine pellet were similar to AUCs in full void urine of an independent previous cohort. GHSR/MAL reached an AUC of 0.89 (95% confidence interval [CI] 0.84-0.94), at 80% sensitivity and 93% specificity. Sensitivity of GHSR/MAL increased with higher tumour grades, higher tumour stages, in primary vs. recurrent tumours, and in males vs. females.
This technical validation supports the robustness of DNA methylation analysis in urine pellet and full void urine for the non-invasive detection of bladder cancer. Subsequent preclinical validation confirmed the diagnostic potential of GHSR/MAL. These findings underline the diagnostic potential of the marker panel GHSR/MAL for future bladder cancer diagnostics.
开发准确的尿生物标志物，用于非侵入性和具有成本效益的原发性和复发性膀胱癌检测，这被认为是膀胱癌诊断中的主要临床需求之一。本研究的目的是：(1) 通过确定 9 个尿沉渣甲基化标志物与全尿相比的诊断性能，验证先前技术比较的结果；(2) 在临床前环境中验证先前探索性研究中最佳标志物组合 GHSR/MAL 的诊断性能。
前瞻性收集 108 例膀胱癌患者和 100 例年龄和性别匹配的对照者的尿样进行甲基化分析。采用定量甲基化特异性 PCR 法检测尿沉渣中 FAM19A4、GHSR、MAL、miR-129、miR-935、PHACTR3、PRDM14、SST 和 ZIC1 标志物的尿甲基化水平。计算各标志物及 GHSR/MAL 标志物组合的曲线下面积(AUC)。采用卡方检验在总研究人群和不同膀胱癌患者亚组中评估 GHSR/MAL 标志物组合的诊断性能。采用留一法交叉验证评估诊断准确性。
9 种尿甲基化标志物(FAM19A4、GHSR、MAL、miR-129、miR-935、PHACTR3、PRDM14、SST 和 ZIC1)在膀胱癌患者的尿中均显著高于对照组(p<0.001)。在独立的先前队列中，在全尿中检测的 9 个甲基化标志物的 AUC 与在尿沉渣中的 AUC 相似。GHSR/MAL 的 AUC 为 0.89(95%置信区间[CI]0.84-0.94)，灵敏度为 80%，特异性为 93%。GHSR/MAL 的灵敏度随肿瘤分级、肿瘤分期、初发 vs 复发肿瘤和男性 vs 女性的增高而升高。
本技术验证支持尿沉渣和全尿中 DNA 甲基化分析用于非侵入性膀胱癌检测的稳健性。随后的临床前验证证实了 GHSR/MAL 的诊断潜力。这些发现强调了 GHSR/MAL 标志物组合用于未来膀胱癌诊断的诊断潜力。
