title: Androgen receptor mutations are associated with altered epigenomic programming as evidenced by HOXA5 methylation.
abstract: Male external genital differentiation is accompanied by implementation of a long-term, male-specific gene expression pattern indicating androgen programming in cultured genital fibroblasts. We hypothesized the existence of an epigenetic background contributing to this phenomenon. DNA methylation levels in 2 normal scrotal fibroblast strains from 46,XY males compared to 2 labia majora fibroblast strains from 46,XY females with complete androgen insensitivity syndrome (AIS) due to androgen receptor (AR) mutations were analyzed by Illumina GoldenGate methylation arrays®. Results were validated with pyrosequencing in labia majora fibroblast strains from fifteen 46,XY patients and compared to nine normal male scrotal fibroblast strains. HOXA5 showed a significantly higher methylation level in complete AIS. This finding was confirmed by bisulfite pyrosequencing of 14 CpG positions within the HOXA5 promoter in the same strains. Extension of the 2 groups revealed a constant low HOXA5 methylation pattern in the controls in contrast to a highly variable methylation pattern in the AIS patients. HOXA5 represents a candidate gene of androgen-mediated promoter methylation. The constantly low HOXA5 DNA methylation level of normal male scrotal fibroblast strains and the frequently high methylation levels in labia majora fibroblast strains in AIS indicate for the first time that androgen programming in sexual differentiation is not restricted to global gene transcription but also occurs at the epigenetic level.
authors.affiliation: Institute of Human Genetics, Christian Albrechts University of Kiel and University Hospital Schleswig-Holstein, Campus Kiel, Kiel, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Bens
authors.forename: O
authors.initials: O
authors.lastname: Ammerpohl
authors.forename: J I
authors.initials: JI
authors.lastname: Martin-Subero
authors.forename: M
authors.initials: M
authors.lastname: Appari
authors.forename: J
authors.initials: J
authors.lastname: Richter
authors.forename: O
authors.initials: O
authors.lastname: Hiort
authors.forename: R
authors.initials: R
authors.lastname: Werner
authors.forename: F G
authors.initials: FG
authors.lastname: Riepe
authors.forename: R
authors.initials: R
authors.lastname: Siebert
authors.forename: P-M
authors.initials: PM
authors.lastname: Holterhus
mesh_terms: D019175:DNA Methylation / Q000235:genetics; D044127:Epigenesis, Genetic / Q000235:genetics*; D005260:Female; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C489213:HOXA5 protein, human; D018398:Homeodomain Proteins; D011944:Receptors, Androgen