Cancer Center Amsterdam, Department of Pathology, VU University Medical Center, Amsterdam, the Netherlands.
Department of Epidemiology and Biostatistics, Amsterdam Public Health Research Institute, VU University Medical Center, Amsterdam, the Netherlands.
Clin Cancer Res. 2018 Jul 15;24(14):3456-3464. doi: 10.1158/1078-0432.CCR-17-3615. Epub 2018 Apr 9.
Offering self-sampling of cervico-vaginal material for high-risk human papillomavirus (hrHPV) testing is an effective method to increase the coverage in cervical screening programs. Molecular triage directly on hrHPV-positive self-samples for colposcopy referral opens the way to full molecular cervical screening. Here, we set out to identify a DNA methylation classifier for detection of cervical precancer (CIN3) and cancer, applicable to lavage and brush self-samples. We determined genome-wide DNA methylation profiles of 72 hrHPV-positive self-samples, using the Infinium Methylation 450K Array. The selected DNA methylation markers were evaluated by multiplex quantitative methylation-specific PCR (qMSP) in both hrHPV-positive lavage ( = 245) and brush ( = 246) self-samples from screening cohorts. Subsequently, logistic regression analysis was performed to build a DNA methylation classifier for CIN3 detection applicable to self-samples of both devices. For validation, an independent set of hrHPV-positive lavage ( = 199) and brush ( = 287) self-samples was analyzed. Genome-wide DNA methylation profiling revealed 12 DNA methylation markers for CIN3 detection. Multiplex qMSP analysis of these markers in large series of lavage and brush self-samples yielded a 3-gene methylation classifier ( and ). This classifier showed a very good clinical performance for CIN3 detection in both lavage (AUC = 0.88; sensitivity = 74%; specificity = 79%) and brush (AUC = 0.90; sensitivity = 88%; specificity = 81%) self-samples in the validation set. Importantly, all self-samples from women with cervical cancer scored DNA methylation-positive. By genome-wide DNA methylation profiling on self-samples, we identified a highly effective 3-gene methylation classifier for direct triage on hrHPV-positive self-samples, which is superior to currently available methods. .
提供宫颈阴道标本的自我采样进行高危型人乳头瘤病毒（hrHPV）检测是增加宫颈筛查计划覆盖范围的有效方法。对 hrHPV 阳性的自我样本进行直接的分子分类，以确定是否需要转诊行阴道镜检查，这为全面的分子宫颈筛查开辟了道路。在这里，我们旨在为检测宫颈前癌（CIN3）和宫颈癌确定一个适用于冲洗和刷取自我样本的 DNA 甲基化分类器。我们使用 Infinium Methylation 450K 阵列确定了 72 例 hrHPV 阳性的自我样本的全基因组 DNA 甲基化谱。通过多重定量甲基化特异性 PCR（qMSP）在来自筛查队列的 hrHPV 阳性冲洗（n = 245）和刷取（n = 246）自我样本中评估了选定的 DNA 甲基化标记物。随后，进行逻辑回归分析以构建适用于两种设备的自我样本的 CIN3 检测的 DNA 甲基化分类器。为了验证，分析了一组独立的 hrHPV 阳性冲洗（n = 199）和刷取（n = 287）自我样本。全基因组 DNA 甲基化谱分析显示 12 个用于 CIN3 检测的 DNA 甲基化标记物。对这些标记物在大量冲洗和刷取自我样本中的多重 qMSP 分析产生了一个 3 个基因甲基化分类器（和）。该分类器在验证集中的冲洗样本（AUC = 0.88；灵敏度 = 74%；特异性 = 79%）和刷取样本（AUC = 0.90；灵敏度 = 88%；特异性 = 81%）中对 CIN3 检测具有非常好的临床性能。重要的是，所有宫颈癌患者的自我样本均显示 DNA 甲基化阳性。通过对自我样本进行全基因组 DNA 甲基化谱分析，我们确定了一种高度有效的 3 个基因甲基化分类器，可直接对 hrHPV 阳性的自我样本进行分类，优于目前可用的方法。
