FAM19A4 methylation analysis in self-samples compared with cervical scrapes for detecting cervical (pre)cancer in HPV-positive women

Abstract

Background:High-risk human papillomavirus (hrHPV)-positive women require triage to identify those with cervical high-grade intraepithelial neoplasia and cancer (⩾CIN3 (cervical intraepithelial neoplasia grade 3)). FAM19A4 methylation analysis, which detects advanced CIN and cancer, is applicable to different sample types. However, studies comparing the performance of FAM19A4 methylation analysis in hrHPV-positive self-samples and paired physician-taken scrapes are lacking.Methods:We compared the performance of FAM19A4 methylation analysis (and/or HPV16/18 genotyping) in self-samples and paired physician-taken scrapes for ⩾CIN3 detection in hrHPV-positive women (n=450,18–66 years).Results:Overall FAM19A4 methylation levels between sample types were significantly correlated, with strongest correlation in women with ⩾CIN3 (Spearman's ρ 0.697, P<0.001). The performance of FAM19A4 methylation analysis and/or HPV16/18 genotyping did not differ significantly between sample types. In women ⩾30 years, ⩾CIN3 sensitivity of FAM19A4 methylation analysis was 78.4% in self-samples and 88.2% in scrapes (ratio 0.89; CI: 0.75–1.05). In women <30 years, ⩾CIN3 sensitivities were 37.5% and 45.8%, respectively (ratio 0.82; CI: 0.55–1.21). In both groups, ⩾CIN3 specificity of FAM19A4 methylation analysis was significantly higher in self-samples compared with scrapes.Conclusions:FAM19A4 methylation analysis in hrHPV-positive self-samples had a slightly lower sensitivity and a higher specificity for ⩾CIN3 compared with paired physician-taken scrapes. With a similarly good clinical performance in both sample types, combined FAM19A4 methylation analysis and HPV16/18 genotyping provides a feasible triage strategy for hrHPV-positive women, with direct applicability on self-samples.

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

“…Cervical cancer development after a persistent infection with high‐risk HPV is driven by additional host cell changes such as altered DNA methylation 12, 17, 20, 21. In earlier work, we have shown that methylation assays targeting FAM19A4 and/or mir124‐2 genes have competitive performance against other triage options 22, 23, 24, 25, 26. In cross‐sectional and short‐term clinical follow‐up studies among both cervical screening and gynecologic outpatient populations, FAM19A4 methylation analysis has shown a similar sensitivity for cervical intraepithelial neoplasia lesions or worse (CIN3+) as compared to cytology 22, 24.…”

“…In cross‐sectional and short‐term clinical follow‐up studies among both cervical screening and gynecologic outpatient populations, FAM19A4 methylation analysis has shown a similar sensitivity for cervical intraepithelial neoplasia lesions or worse (CIN3+) as compared to cytology 22, 24. Methylation levels of these genes in cervical scrapes increase with the severity of underlying cervical lesion 22, 25. Of interest, FAM19A4 methylation analysis detects (virtually) all cervical carcinomas, and has proven to be more sensitive than cytology for the detection of CIN3 lesions with a longer duration of existence 27.…”

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]. Methylation levels of CADM1, MAL, mir124-2, and FAM19A4 are extraordinarily high in cervical scrapes of women with carcinomas [96,101], resulting in a 100% sensitivity for carcinomas of methylation assays targeting these genes, even when very high thresholds are used to define methylation positivity.…”

“…The combination of Pap cytology and methylation analysis, which have been suggested to serve as complementary markers in terms of detection of early and advanced CIN [32], could serve as a sensitive strategy that, in contrast to Pap cytology alone, detects all carcinomas and virtually all CIN3. Table 5 provides an overview of test characteristics (that is, sensitivity, specificity, NPV, PPV) of Pap cytology, HPV16/18 genotyping, p16/Ki-67 dual-stained cytology, and methylation analysis in a single (outpatient) study population [51,67,100,114]. A general limitation of microscopy-based strategies is the fact that these can only be adequately performed on cervical scrapes collected by health-care professionals.…”

Human Papillomavirus Research: Where Should We Place Our Bets?

“…Methylation testing may be an effective triage tool to detect and characterise women at high risk of developing CIN3. The effectiveness of such tests in predicting CIN3 may influence the screening process in many ways, for example by providing an objective method to reach more accurate prognoses or by helping to avoid overtreatment of women with non-progressive lesions [28-31]. …”