Evaluation of Six Methylation Markers Derived from Genome-Wide Screens for Detection of Cervical Precancer and Cancer

Abstract

Aim: To evaluate the triage performance of six host-cell DNA methylation markers derived from two genome-wide discovery screens for detection of cervical precancer (cervical intraepithelial neoplasia 3 [CIN]) and cancer. Materials & methods: Human papillomavirus-positive cervical scrapes of controls (≤CIN1; n = 352) and women diagnosed with CIN3 (n = 175) or cervical cancer (n = 50) were analyzed for methylation of ASCL1, LHX8, ST6GALNAC5, GHSR, SST and ZIC1. Results: Methylation levels increased significantly with disease severity (all markers p < 0.001). Three markers (ASCL1, LHX8, ZIC1) showed receiver operating characteristic curves with area under the curve >0.800 after leave-one-out cross-validation. Bi-marker panel ASCL1/LHX8 had highest area under the curve (0.882), and detected 83.4% of CIN3 and all cervical cancers at specificity of 82.4%. Conclusion: All six methylation markers showed an equivalent, high performance for the triage of human papillomavirus-positive women using cervical scrapes with complementarity between markers.

Keywords::

• ASCL1
• cervical cancer
• cervical intraepithelial neoplasia
• cervical screening
• DNA methylation markers
• human genome methylation
• human papillomavirus
• LHX8
• triage testing

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at: www.tandfonline.com/doi/suppl/10.2217/epi-2019-0331

Financial & competing interest disclosure

This project was supported by grants of the Dutch Cancer Society (KWF VU 2014-7238; KWF 11337). DAM Heideman, RDM Steenbergen and CJLM Meijer are minority shareholders of self-screen B.V., a spin-off company of VUmc. Self-screen B.V. holds patents related to the work (i.e., high-risk HPV test and methylation markers for cervical screening) and has developed and manufactured HPV and methylation assays, which are licensed to QIAGEN (QIAscreen^® HPV PCR Test and QIAsure^® Methylation Test). DAM Heideman has been on the speakers bureau of QIAGEN and serves occasionally on the scientific advisory boards of Pfizer and Bristol-Myers Squibb. CJLM Meijer has received speakers fee from GSK, QIAGEN and SPMSD/Merck and served occasionally on the scientific advisory board (expert meeting) of GSK, QIAGEN and SPMSD/Merck. CJLM Meijer has a very small number of shares of QIAGEN and holds minority stock in Self-Screen B.V. Until April 2016 he was minority shareholder of Diassay B.V. CJLM Meijer is part-time director of Self-screen B.V. since September 2017. Self-screen B.V. was supported by the Valid-screen project, funded by the SME Instrument in the Horizon 2020 Work Program of the European Commission (Valid-screen 666800). LMA Strooper is currently an employee of Bayer on work that is not related to the manuscript. GBA Wisman is inventor on several patents related to the work, is scientific advisor for CC Diagnostics and has received a grant from CC Diagnostics. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials & methods discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations.

Additional information

Funding

This project was supported by grants of the Dutch Cancer Society (KWF VU 2014-7238; KWF 11337). DAM Heideman, RDM Steenbergen and CJLM Meijer are minority shareholders of self-screen B.V., a spin-off company of VUmc. Self-screen B.V. holds patents related to the work (i.e., high-risk HPV test and methylation markers for cervical screening) and has developed and manufactured HPV and methylation assays, which are licensed to QIAGEN (QIAscreen^® HPV PCR Test and QIAsure^® Methylation Test). DAM Heideman has been on the speakers bureau of QIAGEN and serves occasionally on the scientific advisory boards of Pfizer and Bristol-Myers Squibb. CJLM Meijer has received speakers fee from GSK, QIAGEN and SPMSD/Merck and served occasionally on the scientific advisory board (expert meeting) of GSK, QIAGEN and SPMSD/Merck. CJLM Meijer has a very small number of shares of QIAGEN and holds minority stock in Self-Screen B.V. Until April 2016 he was minority shareholder of Diassay B.V. CJLM Meijer is part-time director of Self-screen B.V. since September 2017. Self-screen B.V. was supported by the Valid-screen project, funded by the SME Instrument in the Horizon 2020 Work Program of the European Commission (Valid-screen 666800). LMA Strooper is currently an employee of Bayer on work that is not related to the manuscript. GBA Wisman is inventor on several patents related to the work, is scientific advisor for CC Diagnostics and has received a grant from CC Diagnostics. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials & methods discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript.