Role ofFAM19A4/miR124-2methylation analysis in predicting regression or non-regression of CIN2/3 lesions: a protocol of an observational longitudinal cohort study

Abstract

IntroductionThe clinical course of high-grade cervical intraepithelial neoplasia (CIN2/3) is characterised by a high spontaneous regression rate. Histological assessment is unable to differentiate between CIN2/3 lesions likely to regress and those likely to persist or progress. Most CIN2/3 lesions are treated by surgical excision, leading to overtreatment of a substantial proportion. In this prospective study, we evaluate the value of DNA methylation of host cell genes, which has shown to be particularly sensitive for the detection of advanced CIN2/3 and cervical cancer, in the prediction of regression or non-regression of CIN2/3 lesions.Methods and analysisThis is a multicentre observational longitudinal study with 24-month follow-up. Women referred for colposcopy with an abnormal cervical scrape, who have been diagnosed with CIN2/3 and a small cervical lesion (≤50% of cervix) will be asked to participate. Participants will be monitored by 6-monthly cytological and colposcopic examination. In case of clinical progression, participants will receive treatment and exit the study protocol. At baseline and during follow-up, self-sampled cervicovaginal brushes and cervical scrapes will be collected for high-risk human papillomavirus (HPV) testing andFAM19A4/miR124-2methylation analysis. A colposcopy-directed biopsy will be taken from all participants at the last follow-up visit. The primary study endpoint is regression or non-regression at the end of the study based on the histological diagnosis. Regression is defined as CIN1 or less. Non-regression is defined as CIN2 or worse. The secondary study endpoint is defined as HPV clearance (double-negative HPV test at two consecutive time-points). The association between methylation status and regression probability will be evaluated by means of χ2testing.Ethics and disseminationEthics approval was obtained in all participating clinics. Results of the main study will be submitted for publication in a peer-reviewed journal.Trial registration numberNTR6069; Pre-results

Classification of high‐grade cervical intraepithelial neoplasia by p16ink4a, Ki‐67, HPV E4 and FAM19A4/miR124‐2 methylation status demonstrates considerable heterogeneity with potential consequences for management

“…23 In addition, a negative FAM19A4/miR124-2 methylation test in HPV-positive women provided a very low 14-year cervical cancer risk. 42 In agreement with the latter finding, preliminary results of the CONCERVE study 22 show that women with CIN2/3 with a negative FAM19A4/miR124-2 methylation test have a higher regression rate than women with a positive methylation test (Kremer, Dick, Meijer et al, unpublished data, manuscript in preparation). Therefore, it is assumed that methylation-positive CIN2/3 lesions have a higher short-term progression risk to cervical cancer than methylationnegative CIN2/3 lesions.…”

“…identification of transforming CIN lesions, as well as the potential to predict regression and/or progression of CIN lesions. 21,22 Furthermore, methylation analysis has demonstrated that CIN2/3 lesions associated with a persistent HPV infection of at least 5 years had cancer-like methylation patterns, suggesting a high short-term progression risk to cervical cancer. 23,24 Methylation analysis in combination with immunohistochemical staining of p16 ink4a , Ki-67 and E4 might provide biomarker profiles that could improve the clinical guidance in women with high-grade CIN to prevent overtreatment of regressive lesions and potential obstetric complications.…”

FAM19A4/miR124-2 methylation analysis as a triage test for HPV-positive women: cross-sectional and longitudinal data from a Dutch screening cohort

“…Indeed, it was recently shown that a methylation panel consisting of host cell and viral genes has the ability to identify progressive CIN2 lesions in young women [28]. Additional studies are presently ongoing to validate these findings [29].…”

The use of host cell DNA methylation analysis in the detection and management of women with advanced cervical intraepithelial neoplasia: a review

“…High methylation of host cell marker EPB41L3 combined with viral methylation markers (S5 classifier) were associated with progression to CIN3. A similar study is ongoing in the Netherlands, evaluating the value of FAM19A4/miR124‐2 methylation in the prediction of regression or non‐regression in untreated women 81 . This study also includes women with CIN3 and women aged up to 55 years.…”