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Bakkum-Gamez, Jamie N.; Sherman, Mark E.; Slettedahl, Seth W.; Mahoney, Douglas W.; Lemens, Maureen A.; Laughlin-Tommaso, Shannon K.; Hopkins, Matthew R.; VanOosten, Ann; Shridhar, Viji; Staub, Julie K.; Cao, Xiaoming; Foote, Patrick H.; Clarke, Megan A.; Burger, Kelli N.; Berger, Calise K.; O'Connell, Maria C.; Doering, Karen A.; Podratz, Karl C.; DeStephano, Christopher C.; Schoolmeester, J. Kenneth; Kerr, Sarah E.; Wentzensen, Nicolas; Taylor, William R.; Kisiel, John B.
Gynecologic oncology, 07/2023, Letnik: 174Journal Article
Alterations in DNA methylation are early events in endometrial cancer (EC) development and may have utility in EC detection via tampon-collected vaginal fluid. For discovery, DNA from frozen EC, benign endometrium (BE), and benign cervicovaginal (BCV) tissues underwent reduced representation bisulfite sequencing (RRBS) to identify differentially methylated regions (DMRs). Candidate DMRs were selected based on receiver operating characteristic (ROC) discrimination, methylation level fold-change between cancers and controls, and absence of background CpG methylation. Methylated DNA marker (MDM) validation was performed using qMSP on DNA from independent EC and BE FFPE tissue sets. Women ≥45 years of age with abnormal uterine bleeding (AUB) or postmenopausal bleeding (PMB) or any age with biopsy-proven EC self-collected vaginal fluid using a tampon prior to clinically indicated endometrial sampling or hysterectomy. Vaginal fluid DNA was assayed by qMSP for EC-associated MDMs. Random forest modeling analysis was performed to generate predictive probability of underlying disease; results were 500-fold in-silico cross-validated. Thirty-three candidate MDMs met performance criteria in tissue. For the tampon pilot, 100 EC cases were frequency matched by menopausal status and tampon collection date to 92 BE controls. A 28-MDM panel highly discriminated between EC and BE (96% (95%CI 89–99%) specificity; 76% (66–84%) sensitivity (AUC 0.88). In PBS/EDTA tampon buffer, the panel yielded 96% (95% CI 87–99%) specificity and 82% (70–91%) sensitivity (AUC 0.91). Next generation methylome sequencing, stringent filtering criteria, and independent validation yielded excellent candidate MDMs for EC. EC-associated MDMs performed with promisingly high sensitivity and specificity in tampon-collected vaginal fluid; PBS-based tampon buffer with added EDTA improved sensitivity. Larger tampon-based EC MDM testing studies are warranted. •Whole methylome sequencing identified novel endometrial cancer (EC) methylated DNA markers (MDMs).•A 28-MDM EC panel discriminated between EC and benign endometrium in vaginal fluid collected by tampons.•An abbreviated 3-MDM EC panel performed with similar sensitivity and specificity as the 28-MDM panel.
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in: SICRIS
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