TY - JOUR
T1 - Identification and validation of a 3-gene methylation classifier for hpv-based cervical screening on self-samples
AU - Verlaat, Wina
AU - Snoek, Barbara C.
AU - Heideman, Danielle A.M.
AU - Wilting, Saskia M.
AU - Snijders, Peter J.F.
AU - Novianti, Putri W.
AU - Van Splunter, Annina P.
AU - Peeters, Carel F.W.
AU - Van Trommel, Nienke E.
AU - Massuger, Leon F.A.G.
AU - Bekkers, Ruud L.M.
AU - Melchers, Willem J.G.
AU - Van Kemenade, Folkert J.
AU - Berkhof, Johannes
AU - Van de Wiel, Mark A.
AU - Meijer, Chris J.L.M.
AU - Steenbergen, Renske D.M.
N1 - Funding Information: We thank Lise De Strooper, Bart Hesselink, Maarten van der Salm, Saskia Doorn, Martijn Bogaarts, and Dénira Agard for excellent technical assistance. In addition, we thank Dr. S. Farkas for providing the raw data of her study (22). This work was supported by the European Research Council (ERC advanced 2012-AdG; 322986; Mass-Care) to C.J.L.M. Meijer and by ZonMw (Netherlands Organisation for Health Research and Development; 91216012) to M.A. van de Wiel. Funding Information: We thank Lise De Strooper, Bart Hesselink, Maarten van der Salm, Saskia Doorn, Martijn Bogaarts, and Denira Agard for excellent technical assistance. In addition, we thank Dr. S. Farkas for providing the raw data of her study (22). This work was supported by the European Research Council (ERC advanced 2012-AdG; 322986; Mass-Care) to C.J.L.M. Meijer and by ZonMw (Netherlands Organisation for Health Research and Development; 91216012) to M.A. van de Wiel. Publisher Copyright: © 2018 American Association for Cancer Research.
PY - 2018/7/15
Y1 - 2018/7/15
N2 - Purpose: Offering self-sampling of cervico-vaginal material for high-risk human papillomavirus (hrHPV) testing is an effective method to increase the coverage in cervical screening programs. Molecular triage directly on hrHPV-positive self-samples for colposcopy referral opens the way to full molecular cervical screening. Here, we set out to identify a DNA methylation classifier for detection of cervical precancer (CIN3) and cancer, applicable to lavage and brush self-samples. Experimental Design: We determined genome-wide DNA methylation profiles of 72 hrHPV-positive self-samples, using the Infinium Methylation 450K Array. The selected DNA methylation markers were evaluated by multiplex quantitative methylation-specific PCR (qMSP) in both hrHPV-positive lavage (n ¼ 245) and brush (n ¼ 246) self-samples from screening cohorts. Subsequently, logistic regression analysis was performed to build a DNA methylation classifier for CIN3 detection applicable to self-samples of both devices. For validation, an independent set of hrHPV-positive lavage (n ¼ 199) and brush (n ¼ 287) self-samples was analyzed. Results: Genome-wide DNA methylation profiling revealed 12 DNA methylation markers for CIN3 detection. Multiplex qMSP analysis of these markers in large series of lavage and brush self-samples yielded a 3-gene methylation classifier (ASCL1, LHX8, and ST6GALNAC5). This classifier showed a very good clinical performance for CIN3 detection in both lavage (AUC ¼ 0.88; sensitivity ¼ 74%; specificity ¼ 79%) and brush (AUC ¼ 0.90; sensitivity ¼ 88%; specificity ¼ 81%) self-samples in the validation set. Importantly, all self-samples from women with cervical cancer scored DNA methylation–positive. Conclusions: By genome-wide DNA methylation profiling on self-samples, we identified a highly effective 3-gene methylation classifier for direct triage on hrHPV-positive self-samples, which is superior to currently available methods.
AB - Purpose: Offering self-sampling of cervico-vaginal material for high-risk human papillomavirus (hrHPV) testing is an effective method to increase the coverage in cervical screening programs. Molecular triage directly on hrHPV-positive self-samples for colposcopy referral opens the way to full molecular cervical screening. Here, we set out to identify a DNA methylation classifier for detection of cervical precancer (CIN3) and cancer, applicable to lavage and brush self-samples. Experimental Design: We determined genome-wide DNA methylation profiles of 72 hrHPV-positive self-samples, using the Infinium Methylation 450K Array. The selected DNA methylation markers were evaluated by multiplex quantitative methylation-specific PCR (qMSP) in both hrHPV-positive lavage (n ¼ 245) and brush (n ¼ 246) self-samples from screening cohorts. Subsequently, logistic regression analysis was performed to build a DNA methylation classifier for CIN3 detection applicable to self-samples of both devices. For validation, an independent set of hrHPV-positive lavage (n ¼ 199) and brush (n ¼ 287) self-samples was analyzed. Results: Genome-wide DNA methylation profiling revealed 12 DNA methylation markers for CIN3 detection. Multiplex qMSP analysis of these markers in large series of lavage and brush self-samples yielded a 3-gene methylation classifier (ASCL1, LHX8, and ST6GALNAC5). This classifier showed a very good clinical performance for CIN3 detection in both lavage (AUC ¼ 0.88; sensitivity ¼ 74%; specificity ¼ 79%) and brush (AUC ¼ 0.90; sensitivity ¼ 88%; specificity ¼ 81%) self-samples in the validation set. Importantly, all self-samples from women with cervical cancer scored DNA methylation–positive. Conclusions: By genome-wide DNA methylation profiling on self-samples, we identified a highly effective 3-gene methylation classifier for direct triage on hrHPV-positive self-samples, which is superior to currently available methods.
UR - http://www.scopus.com/inward/record.url?scp=85050112305&partnerID=8YFLogxK
U2 - https://doi.org/10.1158/1078-0432.CCR-17-3615
DO - https://doi.org/10.1158/1078-0432.CCR-17-3615
M3 - Article
C2 - 29632006
SN - 1078-0432
VL - 24
SP - 3456
EP - 3464
JO - Clinical Cancer Research
JF - Clinical Cancer Research
IS - 14
ER -