Department of Surgery and Cancer, 3rd Floor IRDB, Faculty of Medicine, Imperial College London, Hammersmith Campus, Du Cane Road, London, W12 ONN, London, UK; West London Gynaecology Cancer Centre, Hammersmith Hospital, Imperial Healthcare NHS Trust, UK.
Department of Surgery and Cancer, 3rd Floor IRDB, Faculty of Medicine, Imperial College London, Hammersmith Campus, Du Cane Road, London, W12 ONN, London, UK; Department of Obstetrics and Gynaecology, University of Helsinki and Helsinki University Hospital, Finland.
EBioMedicine. 2019 Dec;50:246-259. doi: 10.1016/j.ebiom.2019.10.053. Epub 2019 Nov 12.
Methylation of viral DNA has been proposed as a novel biomarker for triage of human papillomavirus (HPV) positive women at screening. This systematic review and meta-analysis aims to assess how methylation levels change with disease severity and to determine diagnostic test accuracy (DTA) in detecting high-grade cervical intra-epithelial neoplasia (CIN).
We performed searches in MEDLINE, EMBASE and CENTRAL from inception to October 2019. Studies were eligible if they explored HPV methylation levels in HPV positive women. Data were extracted in duplicate and requested from authors where necessary. Random-effects models and a bivariate mixed-effects binary regression model were applied to determine pooled effect estimates.
44 studies with 8819 high-risk HPV positive women were eligible. The pooled estimates for positive methylation rate in HPV16 L1 gene were higher for high-grade CIN (≥CIN2/high-grade squamous intra-epithelial lesion (HSIL) (95% confidence interval (95%CI:72·7% (47·8-92·2))) vs. low-grade CIN (≤CIN1/low-grade squamous intra-epithelial lesion (LSIL) (44·4% (95%CI:16·0-74·1))). Pooled difference in mean methylation level was significantly higher in ≥CIN2/HSIL vs. ≤CIN1/LSIL for HPV16 L1 (11·3% (95%CI:6·5-16·1)). Pooled odds ratio of HPV16 L1 methylation was 5·5 (95%CI:3·5-8·5) for ≥CIN2/HSIL vs. ≤CIN1/LSIL (p < 0·0001). HPV16 L1/L2 genes performed best in predicting CIN2 or worse (pooled sensitivity 77% (95%CI:63-87), specificity 64% (95%CI:55-71), area under the curve (0·73 (95%CI:0·69-0·77)).
Higher HPV methylation is associated with increased disease severity, whilst HPV16 L1/L2 genes demonstrated high diagnostic accuracy to detect high-grade CIN in HPV16 positive women. Direct clinical use is limited by the need for a multi-genotype and standardised assays. Next-generation multiplex HPV sequencing assays are under development and allow potential for rapid, automated and low-cost methylation testing.
NIHR, Genesis Research Trust, Imperial Healthcare Charity, Wellcome Trust NIHR Imperial BRC, European Union's Horizon 2020.
病毒 DNA 的甲基化已被提议作为 HPV 阳性女性筛查时用于分流的新型生物标志物。本系统评价和荟萃分析旨在评估随着疾病严重程度的变化，甲基化水平如何变化，并确定检测高级别宫颈上皮内瘤变（CIN）的诊断测试准确性（DTA）。
我们从成立到 2019 年 10 月在 MEDLINE、EMBASE 和 CENTRAL 中进行了检索。如果研究探讨了 HPV 阳性女性中 HPV 甲基化水平，则符合入选标准。我们对数据进行了重复提取，如果有必要，还向作者请求了数据。应用随机效应模型和双变量混合效应二分类回归模型来确定汇总效应估计值。
有 44 项研究纳入了 8819 名高危 HPV 阳性女性，符合条件。HPV16 L1 基因中阳性甲基化率的汇总估计值在高级别 CIN（≥CIN2/高级别鳞状上皮内病变（HSIL）（95%置信区间（95%CI：72.7%（47.8-92.2）））中高于低级别 CIN（≤CIN1/低级别鳞状上皮内病变（LSIL）（44.4%（95%CI：16.0-74.1）））。HPV16 L1 中，≥CIN2/HSIL 与≤CIN1/LSIL 之间的平均甲基化水平差异显著更高（11.3%（95%CI：6.5-16.1））。HPV16 L1 甲基化的汇总优势比为 5.5（95%CI：3.5-8.5），用于≥CIN2/HSIL 与≤CIN1/LSIL（p<0.0001）。HPV16 L1/L2 基因在预测 CIN2 或更高级别病变方面表现最佳（汇总敏感性 77%（95%CI：63-87），特异性 64%（95%CI：55-71），曲线下面积（0.73（95%CI：0.69-0.77）））。
较高的 HPV 甲基化与疾病严重程度增加相关，而 HPV16 L1/L2 基因在检测 HPV16 阳性女性中的高级别 CIN 方面表现出较高的诊断准确性。直接临床应用受到需要多基因型和标准化检测的限制。下一代多重 HPV 测序检测正在开发中，并为快速、自动化和低成本的甲基化检测提供了潜力。
英国国家健康研究所（NIHR）、创世纪研究信托基金（Genesis Research Trust）、帝国保健慈善基金会（Imperial Healthcare Charity）、惠康信托基金-英国国家健康研究所（NIHR）帝国 BRC、欧盟地平线 2020 计划。
