Cancer risk stratification of anal intraepithelial neoplasia in HIV-positive men by validated methylation markers associated with progression to cancer

Ramon P van der Zee, Olivier Richel, Carel J M van Noesel, Iuliana Ciocănea-Teodorescu, Annina P van Splunter, Timo J Ter Braak, Mayura Nathan, Tamzin Cuming, Michael Sheaff, Alexander Kreuter, Chris J L M Meijer, Wim G V Quint, Henry J C de Vries, Jan M Prins, Renske D M Steenbergen

Research output: Contribution to journalArticleAcademicpeer-review


BACKGROUND: 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, for which reason 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.

METHODS: 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 six 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 ten anal cancer cases with preceding HGAIN at similar anatomic locations, and compared to the cross-sectional series.

RESULTS: Methylation levels of all genes increased with increasing severity of disease (p<0.05). HGAIN revealed a heterogeneous methylation pattern, with a subset resembling cancer. The gene 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.

CONCLUSION: We validated the accuracy of five methylation markers for the detection of anal (pre-)cancer. High methylation levels in HGAIN were associated with progression to cancer. Therefore, these markers provide a promising tool to identify HGAIN in need of treatment, preventing overtreatment of HGAIN with a low cancer progression risk.

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