Abstract 4917: A four-gene methylation marker panel as triage test in hr-HPV positive patients
Authors: Eijsink Jasper JH , Lendvai Agnes , Deregowski Valerie , Klip Harry G. , Verpooten Gonda , Dehaspe Luc , de Bock Geertruida H. , Hollema Harry , van Criekinge Wim , Schuuring Ed , van der Zee Ate GJ , Wisman Bea 
Published: February 2011

Abstract
Objective: Currently no cervical neoplasia specific methylation markers with high sensitivity and specificity are available for use in population-based screening on (pre)malignant cervical neoplasia. Aim of the study was to identify new methylation markers and to design and validate a methylation marker panel for triage of hr-HPV positive patients.
Methods: Quantitative methylation specific PCRs (MSP) on OpenArray™ platform, representing 424 primers of 213 cancer specific methylated genes, were performed on frozen tissue samples from 84 cervical cancer patients and 106 normal cervices. The top 20 of ranked methylation markers were validated by LightCycler® MSP experiments. Then the top 3 methylation markers based on ROC analysis were selected for further clinical validation in combination with C13ORF18 (previously identified by our group), on cervical scrapings from 74 cervical cancer patients, 69 normal cervices and 148 patients referred with an abnormal Pap smear. Finally, a scenario analysis for population based screening program was performed to compare the diagnostic performance of our methylation panel to conventional liquid based cytology after primary hr-HPV testing.
Results: Three cervical neoplasia specific methylation markers (JAM3, EPB41L3 and TERT) discriminated strongly between cervical scrapings from cervical cancer patients and healthy controls (p&lt;0.0001). Our methylation panel (JAM3, EPB4lL3, TERT and C13ORF18) detected 94% of cervical cancers, 82% CIN3+ and 65% CIN2+, while specificity was 79% for CIN0/1 lesions. Scenario analysis showed that primary hr-HPV testing combined with our methylation marker panel as a triage test resulted in a higher identification of CIN3 and cervical cancers, a higher percentage of correct referrals and less patient-doctor contacts compared to hr-HPV testing in combination with liquid based cytology.
Conclusion: Our study resulted in the discovery of three new cervical neoplasia specific methylation markers. Our methylation panel comprising 4 genes might be an alternative triage test after primary hr-HPV testing and its possible application deserves to be further explored in large population-based screening programs for cervical neoplasia.
This study was supported by OncoMethylome Sciences S.A., Liège, Belgium, by the Dutch Cancer Society (NKB) (project-number RUG 2004-3161) and by the “Direction générale des Technologies, de la Recherche et de l'Énergie” of the Walloon Region of Belgium.
Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4917.