title: Role for tissue-dependent methylation differences in the expression of FOXE1 in nontumoral thyroid glands.
abstract: BACKGROUND
  Discordance of monozygotic twins for thyroid dysgenesis suggests that epigenetic mechanisms may underlie defects in thyroid gland development. This prompted us to evaluate whether differentially methylated regions (DMRs) can be found between human thyroids (either eutopic or ectopic) and matched leukocytes.
  
  
  METHODS
  To compare the genome-wide methylation profile of thyroids and leukocytes, immunoprecipitated methylated DNA was interrogated on human promoter plus CpG island tiling arrays. In addition, the methylation profile of the human FOXE1, PAX8, and NKX2.1 promoter was examined using bisulfite sequencing. Finally, the functional impact of CpG methylation of the promoter on FOXE1 expression was assessed with luciferase assays.
  
  
  RESULTS
  Genome-wide methylation profiling and bisulfite sequencing of CpG islands of PAX8 and NKX2.1 promoters revealed no DMR between thyroid and leukocytes. However, bisulfite sequencing revealed that the methylation level of two consecutive CpG dinucleotides (CpG14 and CpG15, which were not covered by the genome-wide array) in one CpG island of the FOXE1 promoter (-1600 to -1140 from the transcription start site) is significantly higher in leukocytes than in eutopic or ectopic thyroid tissues, suggesting that methylation of this region may decrease FOXE1 gene expression. Indeed, luciferase activities were decreased when FOXE1 promoter constructs were methylated in vitro. Moreover, derepression of luciferase activity was observed when the methylation of CpG14 and CpG15 was prevented by mutations.
  
  
  CONCLUSIONS
  We report a tissue-dependent DMR in the FOXE1 promoter. This DMR contains two consecutive CpG dinucleotides, which are epigenetic modifiers of FOXE1 expression in nontumoral tissues.
authors.affiliation: Endocrinology Service and Research Center (R.A.-K., F.M., C.D., G.V.V., J.D.), Centre Hospitalier Universitaire Sainte Justine, Department of Pediatrics, University of Montréal, Montréal, Canada H3T 1C5; Department of Biochemistry (R.A.-K., J.D.), University of Montréal, Montréal, Canada H3C 3J7; and Endocrinology and Diabetes Unit (J.-P.C.), Department of Pediatrics, British Columbia Children's Hospital, University of British Columbia, Vancouver, Canada V6H 3V4.
authors.forename: Rasha
authors.initials: R
authors.lastname: Abu-Khudir
authors.forename: Fabien
authors.initials: F
authors.lastname: Magne
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Chanoine
authors.forename: Cheri
authors.initials: C
authors.lastname: Deal
authors.forename: Guy
authors.initials: G
authors.lastname: Van Vliet
authors.forename: Johnny
authors.initials: J
authors.lastname: Deladoëy
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D018899:CpG Islands; D019175:DNA Methylation; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D020014:K562 Cells; D007962:Leukocytes / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D011401:Promoter Regions, Genetic; D051381:Rats; D013961:Thyroid Gland / Q000378:metabolism*
chemical_list: C106894:FOXE1 protein, human; D051858:Forkhead Transcription Factors