Detection of hypermethylated genes as markers for cervical screening in women living with HIV

Abstract

IntroductionTo evaluate the performance of hypermethylation analysis of ASCL1,LHX8 and ST6GALNAC5 in physician‐taken cervical scrapes for detection of cervical cancer and cervical intraepithelial neoplasia (CIN) grade 3 in women living with HIV (WLHIV) in South Africa.MethodsSamples from a prospective observational cohort study were used for these analyses. Two cohorts were included: a cohort of WLHIV who were invited for cervical screening (n = 321) and a gynaecologic outpatient cohort of women referred for evaluation of abnormal cytology or biopsy proven cervical cancer (n = 108, 60% HIV seropositive). Cervical scrapes collected from all subjects were analysed for hypermethylation of ASCL1,LHX8 and ST6GALNAC5 by multiplex quantitative methylation specific PCR (qMSP). Histology endpoints were available for all study subjects.ResultsHypermethylation levels of ASCL1,LHX8 and ST6GALNAC5 increased with severity of cervical disease. The performance for detection of CIN3 or worse (CIN3+) as assessed by the area under the receiver operating characteristic (ROC) curves (AUC) was good for ASCL1 and LHX8 (AUC 0.79 and 0.81 respectively), and moderate for ST6GALNAC5 (AUC 0.71). At a threshold corresponding to 75% specificity, CIN3+ sensitivity was 72.1% for ASCL1 and 73.8% for LHX8 and all samples from women with cervical cancer scored positive for these two markers.ConclusionsHypermethylation analysis of ASCL1 or LHX8 in cervical scrape material of WLHIV detects all cervical carcinomas with an acceptable sensitivity and good specificity for CIN3+, warranting further exploration of these methylation markers as a stand‐alone test for cervical screening in low‐resource settings.

DNA Methylation and Hydroxymethylation in Cervical Cancer: Diagnosis, Prognosis and Treatment

“…ASCL1 and LHX8 correlated well with CIN3 + (AUC 0.79 and 0.81, respectively) and ST6GALNAC5 correlated moderately with CIN3 + (AUC 0.71) of ASCL1, LHX8, and ST6GALNAC5 exhibited 72.1, 73.8, and 55.7% sensitivity with CIN3 + , respectively. In conclusion, ASCL1 and LHX8 have better performance and higher sensitivity, thereby making them suitable biomarkers for early diagnosis (Kremer et al, 2018). Interestingly, ASCL1 functions as an oncogenic transcription factor in lung cancer (Lenhart et al, 2015).…”

“…Other reports have not been successful in correlating LHX8 to cancer. However, Kremer et al (2018) have reported increasing levels of hypermethylated ASCL1 and LHX8 with increasing severity of cervical cancer. This could be attributed to contamination by other factors like the human immunodeficiency virus (owing to the infected patient cohort).…”

“…Clarke et al (2017) demonstrated that, compared with healthy controls, ADCYAP1, ASCL1, CADM1, DCC, ATP10, DBC1, HS3ST2, MOS, SOX1, MYOD1, SOX17, and TMEFF2 showed higher levels of methylation. ASCL1, LHX8, and ST6GALNAC5 exhibited increased DNA methylation in cervical cancer; DNA hypermethylation also increases with the severity of cervical cancer (Kremer et al, 2018). Moreover, Verlaat et al showed that DNA methylation usually occurs at the pre-tumorigenic stage and reaches the highest level after tumorigenesis induced by hrHPV.…”

“…Taken together, DcR1 may be a promising biomarker for the early diagnosis of cervical cancer and help select patients requiring early treatment. Kremer et al (2018) have identified three related genes, including ASCL1, LHX8, and ST6GALNAC5. ASCL1 and LHX8 correlated well with CIN3 + (AUC 0.79 and 0.81, respectively) and ST6GALNAC5 correlated moderately with CIN3 + (AUC 0.71) of ASCL1, LHX8, and ST6GALNAC5 exhibited 72.1, 73.8, and 55.7% sensitivity with CIN3 + , respectively.…”

“…This could be attributed to contamination by other factors like the human immunodeficiency virus (owing to the infected patient cohort). However, Kremer et al (2018) also showed that using one of these three genes (ASCL1, LHX8, and ST6GALNAC5) was not sensitive. Verlaat et al (2018a) have described a highly effective 3-gene methylation classifier (ASCL1, LHX8, and ST6GALNAC5) to detect the pre-and tumorigenic stage of cervical cancer.…”