Clinical performance of methylation as a biomarker for cervical carcinoma in situ and cancer diagnosis: A worldwide study

Abstract

The shift towards primary human papillomavirus (HPV)‐based screening has necessitated the search for a secondary triage test that provides sufficient sensitivity to detect high‐grade cervical intraepithelial neoplasia (CIN) and cancer, but also brings an improved specificity to avoid unnecessary clinical work and colposcopy referrals. We evaluated the performance of the previously described DNA‐methylation test (S5) in detecting CIN3 and cancers from diverse geographic settings in high‐, medium‐ and low‐income countries, using the cut‐off of 0.80 and exploratory cut‐offs of 2.62 and 3.70. Assays were performed using exfoliated cervical specimens (n = 808) and formalin‐fixed biopsies (n = 166) from women diagnosed with cytology‐negative results (n = 220), CIN3 (n = 204) and cancer stages I (n = 245), II (n = 249), III (n = 28) and IV (n = 22). Methylation increased proportionally with disease severity (Cuzick test for trend, P < .0001). S5 accurately separated women with negative‐histology from CIN3 or cancer (P < .0001). At the 0.80 cut‐off, 543/544 cancers were correctly identified as S5 positive (99.81%). At cut‐off 3.70, S5 showed a sensitivity of 95.77% with improved specificity. The S5 odds ratios of women negative for cervical disease vs CIN3+ were significantly higher than for HPV16/18 genotyping at all cut‐offs (all P < .0001). At S5 cut‐off 0.80, 96.15% of consistently high‐risk human papillomavirus (hrHPV)‐negative cancers (tested with multiple hrHPV‐genotyping assay) were positive by S5. These cancers may have been missed in current primary hrHPV‐screening programmes. The S5 test can accurately detect CIN3 and malignancy irrespective of geographic context and setting. The test can be used as a screening and triage tool. Adjustment of the S5 cut‐off can be performed considering the relative importance given to sensitivity vs specificity.

DNA methylation testing with S5 for triage of high‐risk HPV positive women

“…Although no statewide organised screening program exists, coverage has been shown to be reasonably high. 32 Inclusion criteria included women whose screening cytology immediately preceding their biopsy was hrHPV positive for one or more of 13 hrHPV genotypes (HPV16, 18,31,33,35,39,45,51,52,56,58,59 and 68; N = 4112). Cytology and histology classifications were obtained from the New Mexico HPV Pap Registry (NMHPVPR) and were based on community laboratory results and pathologist diagnoses.…”

Triage performance and predictive value of the human gene methylation panel among women positive on self‐collected HPV test: Results from a prospective cohort study

“…Assays detecting methylation of HPV DNA and/or human genes have a lot of potential as a triaging test for self-collected samples. [8][9][10] DNA hypermethylation of the viral upstream regulatory region (URR) was shown to induce HPV oncoproteins E6 and E7 overexpression, the driving force of a transforming HPV infection. 11 After persistent infection, overexpressed E6 and E7 alter methylation in host cell DNA.…”

“…There is growing interest to evaluate various biomarkers as the reflex triage test on self‐collected samples without recalling the women. Assays detecting methylation of HPV DNA and/or human genes have a lot of potential as a triaging test for self‐collected samples 8‐10 . DNA hypermethylation of the viral upstream regulatory region (URR) was shown to induce HPV oncoproteins E6 and E7 overexpression, the driving force of a transforming HPV infection 11 .…”

Cervical cancer, geographical inequalities, prevention and barriers in resource depleted countries (Review)

“…The sensitivity and specificity for these markers in identification of CINIII were 78.85 and 73.55% respectively ( 80 ). It is worth noting that methylation marker studies aside from being very specific and sensitive, they can accurately detect CIN3 and malignancy irrespective of geographic context and setting ( 81 ).…”