A promising DNA methylation signature for the triage of high-risk human papillomavirus DNA-positive women

• Published in: PloS one 2014-03, Vol.9 (3), p.e91905-e91905
• Main Authors: Hansel, Alfred, Steinbach, Daniel, Greinke, Christiane, Schmitz, Martina, Eiselt, Juliane, Scheungraber, Cornelia, Gajda, Mieczyslaw, Hoyer, Heike, Runnebaum, Ingo B, Dürst, Matthias
• Format: Article
• Language: English
• Subjects: Age; Biology and life sciences; Biomarkers, Tumor - genetics; Cancer; Cancer genetics; Cancer screening; Cellular biology; Cervical cancer; Cervical carcinoma; Cervix; Cervix Uteri - pathology; Cervix Uteri - virology; Comparative analysis; CpG islands; CpG Islands - genetics; Cross-Sectional Studies; Cytology; Deoxyribonucleic acid; DNA; DNA methylation; DNA Methylation - genetics; DNA microarrays; DNA testing; DNA, Viral - genetics; DNA, Viral - isolation & purification; Dysplasia; Epigenetics; Experiments; Female; Genes; Genetic Markers; Genome, Human - genetics; Genomes; Gynecology; Human papillomavirus; Humans; Hybridization; Lesions; Markers; Medical diagnosis; Medical research; Medical screening; Medicine and Health Sciences; Methylation; Papillomaviridae - genetics; Papillomavirus; Papillomavirus infections; Polymerase Chain Reaction; Quality; Reproducibility of Results; Risk Factors; Tissues; Triage; Uterine Cervical Neoplasms - diagnosis; Uterine Cervical Neoplasms - virology; Virology; Womens health
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0091905

High-risk human papillomavirus (hrHPV)-DNA testing is frequently performed parallel to cytology for the detection of high-grade dysplasia and cervical cancer particularly in women above 30 years of age. Although highly sensitive, hrHPV testing cannot distinguish between HPV-positive women with or without clinically relevant lesions. However, in principle discrimination is possible on the basis of DNA methylation markers. In order to identify novel DNA regions which allow an effective triage of hrHPV-positive cases, hypermethylated DNA enriched from cervical cancers was compared with that from cervical scrapes of HPV16-positive cases with no evidence for disease by CpG island microarray hybridization. The most promising marker regions were validated by quantitative methylation-specific PCR (qMSP) using DNA from archived cervical tissues and cervical scrapes. The performance of these markers was then determined in an independent set of 217 hrHPV-positive cervical scrapes from outpatients with histopathological verification. A methylation signature comprising the 5' regions of the genes DLX1, ITGA4, RXFP3, SOX17 and ZNF671 specific for CIN3 and cervical cancer (termed CIN3+) was identified and validated. A high detection rate of CIN3+ was obtained if at least 2 of the 5 markers were methylated. In the subsequent cross-sectional study all cervical carcinomas (n = 19) and 56% (13/23) of CIN3 were identified by this algorithm. Only 10% (11/105) of hrHPV-positive women without histological evidence of cervical disease were scored positive by the methylation assay. Of note is that the detection rate of CIN3 differed between age groups. Eight of nine CIN3 were detected among women ≥30 years of age but only five of fourteen among