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

Abstract

Methylation of host and viral genes is promising for triage of women with high‐risk human papillomavirus infections (hrHPV). Using a population‐based sample of hrHPV positive women with cervical biopsies within 12 months after cervical screening, the clinical value of the S5 methylation classifier (S5), HPV genotyping and cytology were compared as potential triage tests, for outcomes of cervical intraepithelial neoplasia (CIN) grade 3 or greater (CIN3+), CIN2+ and CIN2, and the area under the curve (AUC) calculated. S5 scores increased with histopathology severity (Ptrend < .001). For CIN3+, the AUC was 0.780 suggesting S5 provides good discrimination between <CIN3 and CIN3+. AUCs were significant for all pairwise comparisons of <CIN2, CIN2 and CIN3+ (P < .001). The positive predictive value (PPV) of HPV16/18 genotyping for women with any abnormal cytology was greater than S5 (25.36% vs 20.87%, P = .005) for CIN3+, while sensitivity was substantially greater for S5 (83.33% vs 59.28%, P < .001). Restricting to women with abnormal cytology, but excluding those with high‐grade cytology, both S5 and HPV16/18 provided CIN3+ PPVs high enough to recommend colposcopy. Triage with S5 also appeared useful for hrHPV positive women negative for HPV16/18 (CIN3+ PPV: 7.33%, sensitivity: 57.52%). S5 provided increased sensitivity for CIN3+ compared to HPV16/18 genotyping for hrHPV positive women, overall and when restricted to women with abnormal cytology, suggesting S5 may improve colposcopy referral. S5 also has the ability to distinguish between <CIN2, CIN2 and CIN3+, a finding of importance for managing CIN2, given the complexity and uncertainty associated with this diagnosis.

Long‐term prediction by DNA methylation of high‐grade cervical intraepithelial neoplasia: Results of the ARTISTIC cohort

“…Elevation of S5 DNA methylation correlates directly with increasing severity of disease and inversely with lead time to diagnosis (Figure 2). S5 methylation testing has 92.8% (95CI: 85.7–97.0) sensitivity for detecting prevalent CIN3 and 100% sensitivity (95%CI: 66.4–100) for detecting prevalent ICC (based on 97 women with CIN3 and 9 with ICC), which is consistent with earlier reports of S5 methylation performance 15,18–20,30,31 . The accumulating results thus indicate that as a triage test, S5 would miss a few prevalent CIN3s and almost no invasive cancers.…”

Assessing the risk of cervical neoplasia in the post‐HPV vaccination era

“…The S5 has been substantially evaluated as a triage test for hrHPV‐positive nonvaccinated women and has demonstrated improved triage performance compared to hrHPV genotyping or cytology alone or combined. 40 , 41 , 42 , 43 These observations suggest that the S5 classifier could help identify women with a high short‐term risk of progression to cancer who need immediate treatment. Hernandez et al 41 suggested that the S5 classifier could reduce colposcopy referrals by 30% to 50% without affecting sensitivity for CIN2+ and CIN3+, therefore significantly improving cost‐effectiveness to allow identification of women with a true risk of cancer.…”

“…In addition, S5 had the ability to distinguish between <CIN2, CIN2 and CIN3+, a finding of importance for managing CIN2, given the complexity and uncertainty associated with this diagnosis. 43 …”

HPV Testing and its Role in Cervical Cancer Screening

“…This assay has shown promise for the detection of CIN2+ and CIN3+ in many studies conducted in several different countries. [164][165][166][167][168][169] Other Assays As an expression of E6 and E7 oncoproteins is much higher in cervical precancers compared with transient infections, there has been interest in measuring these proteins to detect transforming HPV infections. Specifically, a lateral flow assay was developed to target the HPV E6 oncoprotein from HPV types 16 and 18 (OncoE6; Arbor Vita Corporation) that may have triage applications in low-resource settings.…”

“…Several studies have also evaluated a triage classifier assay (“S5 classifier”) based on DNA methylation of the late regions of HPV16, HPV18, HPV31, and HPV33 combined with the promoter region of a human gene EPB41L3. This assay has shown promise for the detection of CIN2+ and CIN3+ in many studies conducted in several different countries 164–169 …”