Performance of DNA methylation analysis of ASCL1, LHX8, ST6GALNAC5, GHSR, ZIC1 and SST for the triage of HPV-positive women: Results from a Dutch primary HPV-based screening cohort

Lisanne Verhoef, Maaike C.G. Bleeker, Nicole Polman, Renske D.M. Steenbergen, Chris J.L.M. Meijer, Willem J.G. Melchers, Ruud L. Bekkers, Anco C. Molijn, Wim G. Quint, Folkert J. van Kemenade, Johannes Berkhof, Daniëlle A.M. Heideman*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Abstract

Methylation of host-cell deoxyribonucleic acid (DNA) has been proposed as a promising biomarker for triage of high-risk (hr) human papillomavirus (HPV) positive women at screening. Our study aims to validate recently identified host-cell DNA methylation markers for triage in an hrHPV-positive cohort derived from primary HPV-based cervical screening in The Netherlands. Methylation markers ASCL1, LHX8, ST6GALNAC5, GHSR, ZIC1 and SST were evaluated relative to the ACTB reference gene by multiplex quantitative methylation-specific PCR (qMSP) in clinician-collected cervical samples (n = 715) from hrHPV-positive women (age 29-60 years), who were enrolled in the Dutch IMPROVE screening trial (NTR5078). Primary clinical end-point was cervical intraepithelial neoplasia grade 3 (CIN3) or cancer (CIN3+). The single-marker and bi-marker methylation classifiers developed for CIN3 detection in a previous series of hrHPV-positive clinician-collected cervical samples were applied. The diagnostic accuracy was visualised using receiver operating characteristic (ROC) curves and assessed through area under the ROC curve (AUC). The performance of the methylation markers to detect CIN3+ was determined using the predefined threshold calibrated at 70% clinical specificity. Individual methylation makers showed good performance for CIN3+ detection, with highest AUC for ASCL1 (0.844) and LHX8 (0.830). Combined as a bi-marker panel with predefined threshold, ASCL1/LHX8 yielded a CIN3+ sensitivity of 76.9% (70/91; 95% CI 68.3-85.6%) at a specificity of 74.5% (465/624; 95% CI 71.1-77.9%). In conclusion, our study shows that the individual host-cell DNA methylation classifiers and the bi-marker panel ASCL1/LHX8 have clinical utility for the detection of CIN3+ in hrHPV-positive women invited for routine screening.

Original languageEnglish
Pages (from-to)440-449
Number of pages10
JournalInternational Journal of Cancer
Volume150
Issue number3
Early online date24 Sept 2021
DOIs
Publication statusPublished - 1 Feb 2022

Bibliographical note

Funding Information:
This work is dedicated to our colleague Prof. Dr. P.J.F. Snijders, who passed away on May 27, 2018. The authors thank all women who participated in the IMPROVE study; the screening organisations Midden-West, Zuid-West, and Oost; the National Institute for Public Health and the Environment; and the physicians, gynaecologists, and pathologists in the study regions. They also thank Daisy Hoek for technical assistance and Mark van de Wiel and Ivonne Martin for statistical support. The authors would like to acknowledge the nationwide network and registry of histo- and cytopathology in the Netherlands (PALGA) for providing data and the Amsterdam UMC Biobank for their high-quality storage of collected samples. The graphical abstract accompanying this article was created with BioRender.com. J.B. had financial support from the European Commission (RISCC, grant number 847845).

Funding Information:
This work is dedicated to our colleague Prof. Dr. P.J.F. Snijders, who passed away on May 27, 2018. The authors thank all women who participated in the IMPROVE study; the screening organisations Midden‐West, Zuid‐West, and Oost; the National Institute for Public Health and the Environment; and the physicians, gynaecologists, and pathologists in the study regions. They also thank Daisy Hoek for technical assistance and Mark van de Wiel and Ivonne Martin for statistical support. The authors would like to acknowledge the nationwide network and registry of histo‐ and cytopathology in the Netherlands (PALGA) for providing data and the Amsterdam UMC Biobank for their high‐quality storage of collected samples. The graphical abstract accompanying this article was created with BioRender.com . J.B. had financial support from the European Commission (RISCC, grant number 847845).

Publisher Copyright:
© 2021 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.

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