Department of Pathology, Amsterdam University Medical Centers, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, Amsterdam, The Netherlands.
Department of Internal Medicine, Division of Infectious Diseases, Amsterdam University Medical Centers, University of Amsterdam, Amsterdam Institute for Infection and Immunity (AI&II), Amsterdam, The Netherlands.
Clin Infect Dis. 2021 Jun 15;72(12):2154-2163. doi: 10.1093/cid/ciaa397.
High-grade anal intraepithelial neoplasia (HGAIN; AIN2-3) is highly prevalent in HIV+ men, but only a minority of these lesions progress towards cancer. Currently, cancer progression risk cannot be established; therefore, no consensus exists on whether HGAIN should be treated. This study aimed to validate previously identified host cell DNA methylation markers for detection and cancer risk stratification of HGAIN.
A large independent cross-sectional series of 345 anal cancer, AIN3, AIN2, AIN1, and normal control biopsies of HIV+ men was tested for DNA methylation of 6 genes using quantitative methylation-specific PCR. We determined accuracy for detection of AIN3 and cancer (AIN3+) by univariable and multivariable logistic regression analysis, followed by leave-one-out cross-validation. Methylation levels were assessed in a series of 10 anal cancer cases with preceding HGAIN at similar anatomic locations, and compared with the cross-sectional series.
Methylation levels of all genes increased with increasing severity of disease (P < .05). HGAIN revealed a heterogeneous methylation pattern, with a subset resembling cancer. ZNF582 showed highest accuracy (AUC = 0.88) for AIN3+ detection, slightly improved by addition of ASCL1 and SST (AUC = 0.89), forming a marker panel. In the longitudinal series, HGAIN preceding cancer displayed high methylation levels similar to cancers.
We validated the accuracy of 5 methylation markers for the detection of anal (pre-) cancer. High methylation levels in HGAIN were associated with progression to cancer. These markers provide a promising tool to identify HGAIN in need of treatment, preventing overtreatment of HGAIN with a low cancer progression risk.
高级别肛门上皮内瘤变（HGAIN；AIN2-3）在 HIV+ 男性中非常普遍，但这些病变只有少数会发展为癌症。目前，无法确定癌症进展的风险；因此，对于 HGAIN 是否应该治疗尚无共识。本研究旨在验证先前鉴定的宿主细胞 DNA 甲基化标志物，用于检测和 HGAIN 的癌症风险分层。
使用定量甲基化特异性 PCR 检测 345 例 HIV+ 男性的肛门癌、AIN3、AIN2、AIN1 和正常对照活检的 6 个基因的 DNA 甲基化。我们通过单变量和多变量逻辑回归分析确定了检测 AIN3 和癌症（AIN3+）的准确性，然后进行了留一法交叉验证。在 10 例具有相似解剖位置的先前存在的 HGAIN 的肛门癌病例系列中评估了甲基化水平，并与横断面系列进行了比较。
所有基因的甲基化水平随疾病严重程度的增加而增加（P<0.05）。HGAIN 显示出异质性的甲基化模式，其中一部分与癌症相似。ZNF582 对 AIN3+检测的准确性最高（AUC=0.88），通过添加 ASCL1 和 SST 略有提高（AUC=0.89），形成一个标志物组合。在纵向系列中，癌症前的 HGAIN 显示出与癌症相似的高甲基化水平。
我们验证了 5 个甲基化标志物用于检测肛门（前）癌的准确性。HGAIN 中的高甲基化水平与癌症进展相关。这些标志物为识别需要治疗的 HGAIN 提供了一个有前途的工具，防止对低癌症进展风险的 HGAIN 进行过度治疗。
