Discovery of new methylation markers to improve screening for cervical intraepithelial neoplasia grade 2/3

被引：69
作者：
Boers, A. [1 ]
Wang, R. [1 ]
van Leeuwen, R. W. [1 ]
Klip, H. G. [1 ]
de Bock, G. H. [2 ]
Hollema, H. [3 ]
van Criekinge, W. [4 ]
de Meyer, T. [4 ]
Denil, S. [4 ]
van der Zee, A. G. J. [1 ]
Schuuring, E. [3 ]
Wisman, G. B. A. [1 ]
机构：
[1] Univ Groningen, Univ Med Ctr Groningen, Canc Res Ctr Groningen, Dept Gynecol Oncol, Internal Postal Code DA13,POB 30-001, NL-9700 RB Groningen, Netherlands
[2] Univ Groningen, Univ Med Ctr Groningen, Canc Res Ctr Groningen, Dept Epidemiol, NL-9700 RB Groningen, Netherlands
[3] Univ Groningen, Univ Med Ctr Groningen, Canc Res Ctr Groningen, Dept Pathol, NL-9700 RB Groningen, Netherlands
[4] Univ Ghent, Dept Math Modeling Stat & Bioinformat, B-9000 Ghent, Belgium
来源：
CLINICAL EPIGENETICS | 2016年 / 8卷
关键词：
Cervical cancer screening; Cervical precancerous lesions; Human papillomavirus (HPV); Cervical scraping; MethylCap-seq; DNA methylation; Quantitative methylation-specific PCR (QMSP); HUMAN-PAPILLOMAVIRUS DNA; FOLLOW-UP; PROMOTER HYPERMETHYLATION; POSITIVE WOMEN; TRIAGE TEST; CANCER; RISK; GENOME; CYTOLOGY; SPECIFICITY;
D O I：
10.1186/s13148-016-0196-3
中图分类号：
R73 [肿瘤学];
学科分类号：
100214 ;
摘要：
Background: Assessment of DNA promoter methylation markers in cervical scrapings for the detection of cervical intraepithelial neoplasia (CIN) and cervical cancer is feasible, but finding methylation markers with both high sensitivity as well as high specificity remains a challenge. In this study, we aimed to identify new methylation markers for the detection of high-grade CIN (CIN2/3 or worse, CIN2+) by using innovative genome-wide methylation analysis (MethylCap-seq). We focused on diagnostic performance of methylation markers with high sensitivity and high specificity considering any methylation level as positive. Results: MethylCap-seq of normal cervices and CIN2/3 revealed 176 differentially methylated regions (DMRs) comprising 164 genes. After verification and validation of the 15 best discriminating genes with methylation-specific PCR (MSP), 9 genes showed significant differential methylation in an independent cohort of normal cervices versus CIN2/3 lesions (p < 0.05). For further diagnostic evaluation, these 9 markers were tested with quantitative MSP (QMSP) in cervical scrapings from 2 cohorts: (1) cervical carcinoma versus healthy controls and (2) patients referred from population-based screening with an abnormal Pap smear in whom also HPV status was determined. Methylation levels of 8/9 genes were significantly higher in carcinoma compared to normal scrapings. For all 8 genes, methylation levels increased with the severity of the underlying histological lesion in scrapings from patients referred with an abnormal Pap smear. In addition, the diagnostic performance was investigated, using these 8 new genes and 4 genes (previously identified by our group: C13ORF18, JAM3, EPB41L3, and TERT). In a triage setting (after a positive Pap smear), sensitivity for CIN2+ of the best combination of genes (C13ORF18/JAM3/ANKRD18CP) (74 %) was comparable to hrHPV testing (79 %), while specificity was significantly higher (76 % versus 42 %, p <= 0.05). In addition, in hrHPV-positive scrapings, sensitivity and specificity for CIN2+ of this best-performing combination was comparable to the population referred with abnormal Pap smear. Conclusions: We identified new CIN2/3-specific methylation markers using genome-wide DNA methylation analysis. The diagnostic performance of our new methylation panel shows higher specificity, which should result in prevention of unnecessary colposcopies for women referred with abnormal cytology. In addition, these newly found markers might be applied as a triage test in hrHPV-positive women from population-based screening. The next step before implementation in primary screening programs will be validation in population-based cohorts.
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页数：16
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