CADM1andMALpromoter methylation levels in hrHPV-positive cervical scrapes increase proportional to degree and duration of underlying cervical disease

Abstract

Combined detection of cell adhesion molecule 1 (CADM1) and T-lymphocyte maturation-associated protein (MAL) promoter methylation in cervical scrapes is a promising triage strategy for high-risk human papillomavirus (hrHPV)-positive women. Here, CADM1 and MAL DNA methylation levels were analysed in cervical scrapes of hrHPV-positive women with no underlying high-grade disease, high-grade cervical intraepithelial neoplasia (CIN) and cervical cancer. CADM1 and MAL methylation levels in scrapes were first related to CIN-grade of the corresponding biopsy and second to CIN-grade stratified by the presence of 'normal' or 'abnormal' cytology as present in the accompanying scrape preceding the cervical biopsy. The scrapes included 167 women with CIN1, 54 with CIN2/3 and 44 with carcinoma. In a separate series of hrHPV-positive scrapes of women with CIN2/3 (n 5 48), methylation levels were related to duration of preceding hrHPV infection (PHI; <5 and 5 years). Methylation levels were determined by quantitative methylation-specific PCR and normal cytology scrapes of hrHPV-positive women with histologically CIN1 served as reference. CADM1 and MAL methylation levels increased proportional to severity of the underlying lesion, showing an increase of 5.3-and 6.2-fold in CIN2/3, respectively, and 143.5-and 454.9-fold in carcinomas, respectively, compared to the reference. Methylation levels were also elevated in CIN2/3 with a longer duration of PHI (i.e. 11.5-and 13.6-fold, respectively). Moreover, per histological category, methylation levels were higher in accompanying scrapes with abnormal cytology than in scrapes with normal cytology. Concluding, CADM1 and MAL promoter methylation levels in hrHPV-positive cervical scrapes are related to the degree and duration of underlying cervical disease and markedly increased in cervical cancer.

Host-cell DNA methylation patterns during high-risk HPV-induced carcinogenesis reveal a heterogeneous nature of cervical pre-cancer

“…These results correspond to the finding that only a subset of CIN2/3 will progress to cancer over a long time period [18,19]. Previous studies on copy number changes and DNA methylation levels of only a few genes also showed a cancer-like profile in only a subset of CIN2/3 lesions [11,14,20,21]. These lesions were characterized by a preceding hrHPV infection of >5 years and considered as more advanced lesions, which have a high short-term progression risk to cancer.…”

Cervical cancer risk in HPV‐positive women after a negative FAM19A4/mir124‐2 methylation test: A post hoc analysis in the POBASCAM trial with 14 year follow‐up

“…In fact, FAM19A4 methylation analysis tested positive in all cervical scrapes of women with CIN3 lesions with a duration of the associated HPV infection of >5 years 22. Those lesions are characterized by a ‘cancer‐like’ (epi)genetic profile and have therefore been considered as advanced precancers 22, 28. Based on this feature, triage testing by methylation analysis has been proposed to confer a high reassurance against short‐term risk of cervical cancer 12.…”

Management of high-risk HPV-positive women for detection of cervical (pre)cancer

“…Furthermore, host cell methylation analysis of CADM1, MAL, mir124-2, and FAM19A4 in cervical scrapes allows the discrimination of high-grade CIN with a long-term preceding hrHPV-infection from those with a short-term hrHPV infection [95,96]. Concordantly, in both cervical scrapes and selfsampled cervicovaginal lavages, methylation levels of FAM19A4 have been shown to correspond with the estimated CIN lesion volume [100].…”

“…In both cervical scrapes and lavages the NPVs of CADM1/ MAL and FAM19A4(/mir124-2) methylation analysis (86.3-95.3%) are below the suggested 98% threshold [38]. However, given that methylation analysis of CADM1/MAL and FAM19A4(/mir124-2) has a very high sensitivity for carcinomas and CIN3 with a long-term previous hrHPV-infection [95,96], we argue that methylation-negative CIN3 lesions have a low short-term risk of progression to cancer. From this point of view, one might consider the use of (CADM1/MAL and FAM19A4(/mir124-2)) methylation analysis for triage of hrHPVpositive women accepting a somewhat lower NPV, thereby allowing CIN2/3 with low short-term progression risk to remain undetected.…”

“…Methylation levels of certain host cell genes (such as CADM1, MAL, mir124-2, JAM3, TERT, C13ORF, EPB41L3, ANKRD18CP, GFRA1, CDH6, LHX8, GATA4, PRDM14, and FAM19A4) in cervical scrapes have been shown to increase with the severity of underlying cervical (pre)cancer [51,[95][96][97][98][99]. Furthermore, host cell methylation analysis of CADM1, MAL, mir124-2, and FAM19A4 in cervical scrapes allows the discrimination of high-grade CIN with a long-term preceding hrHPV-infection from those with a short-term hrHPV infection [95,96].…”