Triage of high-risk HPV-positive women in population-based screening by miRNA expression analysis in cervical scrapes; a feasibility study

Abstract

BackgroundPrimary testing for high-risk HPV (hrHPV) is increasingly implemented in cervical cancer screening programs. Many hrHPV-positive women, however, harbor clinically irrelevant infections, demanding additional disease markers to prevent over-referral and over-treatment. Most promising biomarkers reflect molecular events relevant to the disease process that can be measured objectively in small amounts of clinical material, such as miRNAs. We previously identified eight miRNAs with altered expression in cervical precancer and cancer due to either methylation-mediated silencing or chromosomal alterations. In this study, we evaluated the clinical value of these eight miRNAs on cervical scrapes to triage hrHPV-positive women in cervical screening.ResultsExpression levels of the eight candidate miRNAs in cervical tissue samples (n = 58) and hrHPV-positive cervical scrapes from a screening population (n = 187) and cancer patients (n = 38) were verified by quantitative RT-PCR. In tissue samples, all miRNAs were significantly differentially expressed (p < 0.05) between normal, high-grade precancerous lesions (CIN3), and/or cancer. Expression patterns detected in cervical tissue samples were reflected in cervical scrapes, with five miRNAs showing significantly differential expression between controls and women with CIN3 and cancer. Using logistic regression analysis, a miRNA classifier was built for optimal detection of CIN3 in hrHPV-positive cervical scrapes from the screening population and its performance was evaluated using leave-one-out cross-validation. This miRNA classifier consisted of miR-15b-5p and miR-375 and detected a major subset of CIN3 as well as all carcinomas at a specificity of 70%. The CIN3 detection rate was further improved by combining the two miRNAs with HPV16/18 genotyping. Interestingly, both miRNAs affected the viability of cervical cancer cells in vitro.ConclusionsThis study shows that miRNA expression analysis in cervical scrapes is feasible and enables the early detection of cervical cancer, thus underlining the potential of miRNA expression analysis for triage of hrHPV-positive women in cervical cancer screening.Electronic supplementary materialThe online version of this article (10.1186/s13148-018-0509-9) contains supplementary material, which is available to authorized users.

Genome‐wide microRNA analysis of HPV‐positive self‐samples yields novel triage markers for early detection of cervical cancer

“…Moreover, we found that differential expression of the identified miRNAs could be confirmed in independent cervical tissue specimens analyzed by a different method . In addition, recent data show increased expression of miR‐15b in HPV‐positive physician‐taken cervical scrapes from women with CIN3 and SCC . Similarly, preliminary miRNA expression data show consistent results for miR‐20a, miR‐93 and miR‐222 in HPV‐positive cervical scrapes (Supporting Information Fig.…”

“…30 In addition, recent data show increased expression of miR-15b in HPV-positive physician-taken cervical scrapes from women with CIN3 and SCC. 33 Similarly, preliminary miRNA expression data show consistent results for miR-20a, miR-93 and miR-222 in HPV-positive cervical scrapes (Supporting Information Fig. S3).…”

“…Detailed characteristics of physician‐taken cervical scrapes (Supporting Information Fig. S3), including miRNA expression analysis by qPCR have been described previously …”

Complementarity between miRNA expression analysis and DNA methylation analysis in hrHPV-positive cervical scrapes for the detection of cervical disease

“…Analysis of differential microRNA (miRNA) expression for diagnostic and prognostic purposes has gained increasing attention in recent years. Others and we have previously shown that differential miRNA expression between normal and cervical (pre)cancer cannot only be detected in cervical tissue samples but also in cervical exfoliated cells present in cervical scrapes and self-collected cervico-vaginal specimens (selfsamples) [5][6][7][8]. Although the feasibility of miRNA analysis for the triage of hrHPV-positive women has been demonstrated, an optimal miRNA panel for the detection of CIN3 and cervical cancer still remains to be determined [6].…”

“…In this study, the expression levels of 10 candidate miRNAs (miR-15b, miR-125b, miR-149, miR-203a, miR-375, miR-20a, miR-31, miR-93, miR-222, let-7b) derived from an earlier feasibility study on cervical scrapes and a study on hrHPV-positive self-samples [6,8] were determined by qRT-PCR in 209 cervical scrapes of hrHPV-positive women (age: 30-83 years) with known underlying histology that have not been tested for miRNAs before. A predictive miRNA classifier for the detection of CIN3 was built, which was applied to the full sample series and subsequently compared to and combined with the DNA methylation marker FAM19A4 [4].…”