title: Detection of endometrial cancer via molecular analysis of DNA collected with vaginal tampons.
abstract: OBJECTIVE
  We demonstrate the feasibility of detecting EC by combining minimally-invasive specimen collection techniques with sensitive molecular testing.
  
  
  METHODS
  Prior to hysterectomy for EC or benign indications, women collected vaginal pool samples with intravaginal tampons and underwent endometrial brushing. Specimens underwent pyrosequencing for DNA methylation of genes reported to be hypermethylated in gynecologic cancers and recently identified markers discovered by profiling over 200 ECs. Methylation was evaluated individually across CpGs and averaged across genes. Differences between EC and benign endometrium (BE) were assessed using two-sample t-tests and area under the curve (AUC).
  
  
  RESULTS
  Thirty-eight ECs and 28 BEs were included. We evaluated 97 CpGs within 12 genes, including previously reported markers (RASSF1, HSP2A, HOXA9, CDH13, HAAO, and GTF2A1) and those identified in discovery work (ASCL2, HTR1B, NPY, HS3ST2, MME, ADCYAP1, and additional CDH13 CpG sites). Mean methylation was higher in tampon specimens from EC v. BE for 9 of 12 genes (ADCYAP1, ASCL2, CDH13, HS3ST2, HTR1B, MME, HAAO, HOXA9, and RASSF1) (all p<0.05). Among these genes, relative hypermethylation was observed in EC v. BE across CpGs. Endometrial brush and tampon results were similar. Within tampon specimens, AUC was highest for HTR1B (0.82), RASSF1 (0.75), and HOXA9 (0.74). This is the first report of HOXA9 hypermethylation in EC.
  
  
  CONCLUSIONS
  DNA hypermethylation in EC tissues can also be identified in vaginal pool DNA collected via intravaginal tampon. Identification of additional EC biomarkers and refined collection methods are needed to develop an early detection tool for EC.
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA. Electronic address: Bakkum.jamie@mayo.edu.
authors: Jamie N
authors: JN
authors: Bakkum-Gamez
authors: Hormonal and Reproductive Branch (HREB), Division of Cancer Epidemiology and Genetics (DCEG), National Cancer Institute (NCI), USA.
authors: Nicolas
authors: N
authors: Wentzensen
authors: Department of Health Sciences Research, Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA.
authors: Matthew J
authors: MJ
authors: Maurer
authors: Department of Health Sciences Research, Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA.
authors: Kieran M
authors: KM
authors: Hawthorne
authors: Department of Laboratory Medicine and Pathology, Division of Anatomic Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Jesse S
authors: JS
authors: Voss
authors: Department of Laboratory Medicine and Pathology, Division of Anatomic Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Trynda N
authors: TN
authors: Kroneman
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Abimbola O
authors: AO
authors: Famuyide
authors: Department of Laboratory Medicine and Pathology, Division of Anatomic Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Amy C
authors: AC
authors: Clayton
authors: Department of Laboratory Medicine and Pathology, Division of Laboratory Genetics, Mayo Clinic, Rochester, MN, USA.
authors: Kevin C
authors: KC
authors: Halling
authors: Department of Laboratory Medicine and Pathology, Division of Anatomic Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Sarah E
authors: SE
authors: Kerr
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: William A
authors: WA
authors: Cliby
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Sean C
authors: SC
authors: Dowdy
authors: Department of Laboratory Medicine and Pathology, Division of Anatomic Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Benjamin R
authors: BR
authors: Kipp
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Andrea
authors: A
authors: Mariani
authors: Department of Health Sciences Research, Division of Biomedical Statistics and Informatics, Mayo Clinic, Rochester, MN, USA.
authors: Ann L
authors: AL
authors: Oberg
authors: Department of Obstetrics and Gynecology, Division of Gynecologic Surgery, Mayo Clinic, Rochester, MN, USA.
authors: Karl C
authors: KC
authors: Podratz
authors: Department of Laboratory Medicine and Pathology, Division of Experimental Pathology, Mayo Clinic, Rochester, MN, USA.
authors: Viji
authors: V
authors: Shridhar
authors: Hormonal and Reproductive Branch (HREB), Division of Cancer Epidemiology and Genetics (DCEG), National Cancer Institute (NCI), USA.
authors: Mark E
authors: ME
authors: Sherman
mesh_terms: D000368:Aged; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D016022:Case-Control Studies; D018899:CpG Islands; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000235:genetics* / Q000302:isolation & purification*; D016889:Endometrial Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D052576:Menstrual Hygiene Products; D008875:Middle Aged; D009367:Neoplasm Staging; D010865:Pilot Projects; D014621:Vagina / Q000737:chemistry* / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm
keywords: Early detection; Endometrial cancer; Methylation; Tampon; Tao brush