Combined genetic mutations and DNA-methylated genes as biomarkers for endometrial cancer detection from cervical scrapings

Abstract

Background: Endometrial cancer is a common gynecologic cancer. Noninvasive molecular biomarkers for triage of high-risk patients for invasive procedures are needed. Based on the success of cytological Pap smear screening, cervical scrapings are a good source of DNA for molecular testing. In addition to genetic lesions, DNA methylation is a promising biomarker. We assessed the usefulness of combining genetic and epigenetic biomarkers from cervical scrapings to detect endometrial carcinomas. Methods: We performed a retrospective case-control study of 96 consecutive cervical scrapings from patients with abnormal uterine bleeding who underwent surgery for diagnostic evaluation. Thirty and 16 cases were diagnosed with type I and type II endometrial cancers, respectively. The remaining non-cancer cases included normal endometrium (n = 12), benign uterine lesions (n = 20), and endometrial hyperplasia (n = 18). Quantitative methylation-specific PCR and mass spectrometry were used for DNA methylation and genetic mutation analysis. Logistic regression was used to evaluate the clinical performance of these candidate biomarkers. Results: We tested the effectiveness of the methylation status of four genes (BHLHE22, CDO1, TBX5, and HAND2) in endometrial cancer detection. The area under the receiver operating characteristic curves ranged from 0.703 to 0.878, and panels of hypermethylated BHLHE22/CDO1/HAND2 (87.0% sensitivity and 86.0% specificity) and BHLHE22/ CDO1/TBX5 (89.1% sensitivity and 80.0% specificity) showed significant differences and could distinguish benign from malignant endometrial lesions. The sensitivity and specificity in endometrial cancer detection for BHLHE22/ CDO1 were 84.8% and 88.0%, respectively. Both type I and II endometrial carcinomas could be detected using a BHLHE22/CDO1-based methylation profile, suggesting that they may have common epigenomes. Moreover, PTEN and TP53 mutations were found in 63.3% of type I and 93.6% of type II endometrial cancers. Unexpectedly, PTEN and TP53 mutations were commonly found in cervical scrapings of the normal endometrium (25% and 33.3%, respectively) and in cases with benign uterine lesions (10% and 50%, respectively). Finally, combinations of any one mutation of PTEN and TP53 mutations had a sensitivity of 91.3%, but a specificity of only 42.0%. Conclusions: Adding PTEN/TP53 mutation testing to BHLHE22/CDO1-based methylation testing did not improve the detection of endometrial cancer.

Artesunate Suppresses the Proliferation and Development of Estrogen Receptor-α-Positive Endometrial Cancer in HAND2-Dependent Pathway

“…Heart and neural crest derivatives expressed 2 is a transcriptional factor involved in the maintaining of endometrial function ( Kato et al, 2019 ). The methylation and silence of HAND2 contributed to the high risk of EC ( Liew et al, 2019 ; Multinu et al, 2020 ). Based on the immunoprecipitation and pull-down assay, one study also pointed out that HAND2 directly interacted with the amino-terminus of ER-α, resulted in inhibition of the transcriptional expression of it ( Fukuda et al, 2015 ).…”

Targeting Epigenetic Regulators for Endometrial Cancer Therapy: Its Molecular Biology and Potential Clinical Applications

“…However, they demonstrated that a combination of the analysis of copy number, DNA methylation, and gene mutation improved sensitivity (92%). In another study, combining methylation patterns in several genes ( BHLHE22 , CDO1 , TBX5 , and HAND2 ) with mutations in PTEN and TP53 led to an endometrial cancer diagnosis with a sensitivity of 91.3% and a specificity of 42% [ 58 ].…”

Evolutionary Perspective and Expression Analysis of Intronless Genes Highlight the Conservation of Their Regulatory Role

“…Bhlhe22 and Bhlhe23 in turn, are among those that were up-regulated the most in mice during telencephalon development. In humans, Bhlhe22 has been identified as a highly methylated gene in endometrial cancer with potential epigenetic biomarkers in cervical scrapings (Liew et al, 2019), while Bhlhe23 has been linked to mammalian retinal development (Woods et al, 2018).…”