title: Methylome repatterning in a mouse model of Maternal PKU Syndrome.
abstract: Maternal PKU Syndrome (MPKU) is an embryopathy resulting from in utero phenylalanine (PHE) toxicity secondary to maternal phenylalanine hydroxylase deficient phenylketonuria (PKU). Clinical phenotypes in MPKU include mental retardation, microcephaly, in utero growth restriction, and congenital heart defects. Numerous in utero toxic exposures alter DNA methylation in the fetus. The PAH(enu2) mouse is a model of classical PKU while offspring born of hyperphenylalaninemic dams model MPKU. We investigated offspring of PAH(enu2) dams to determine if altered patterns of DNA methylation occurred in response to in utero PHE exposure. As neurologic deficit is the most prominent MPKU phenotype, methylome patterns were assessed in brain tissue using methylated DNA immunoprecipitation and paired-end sequencing. Brain tissues were assessed in E18.5-19 fetuses of PHE unrestricted PAH(enu2) dams, PHE restricted PAH(enu2) dams, and heterozygous(wt/enu2) control dams. Extensive methylome repatterning was observed in offspring of hyperphenylalaninemic dams while the offspring of PHE restricted dams displayed attenuated methylome repatterning. Methylation within coding regions was dominated by noncoding RNA genes. Differential methylation of promoters targeted protein coding genes. To assess the impact of methylome repatterning on gene expression, brain tissue in experimental and control animals were queried with microarrays assessing expression of microRNAs and protein coding genes. Altered expression of methylome-modified microRNAs and protein coding genes was extensive in offspring of hyperphenylalaninemic dams while minimal changes were observed in offspring of PHE restricted dams. Several genes displaying significantly reduced expression have roles in neurological function or genetic disease with neurological phenotypes. These data indicate in utero PHE toxicity alters DNA methylation in the brain which has downstream impact upon gene expression. Altered gene expression may contribute to pathophysiology of neurologic presentation in MPKU.
authors.affiliation: Department of Pathology, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: S F
authors.initials: SF
authors.lastname: Dobrowolski
authors.affiliation: Genomics and Proteomics Core Laboratories, Bioinformatics Core, University of Pittsburgh, 3343 Forbes Avenue, Pittsburgh, PA 15260, USA.
authors.forename: J
authors.initials: J
authors.lastname: Lyons-Weiler
authors.affiliation: Department of Pathology, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: A
authors.initials: A
authors.lastname: Biery
authors.affiliation: Department of Pathology, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: K
authors.initials: K
authors.lastname: Spridik
authors.affiliation: Division of Medical Genetics, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: G
authors.initials: G
authors.lastname: Vockley
authors.affiliation: Department of Pathology, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: E
authors.initials: E
authors.lastname: Kranik
authors.affiliation: Division of Medical Genetics, Children's Hospital of Pittsburgh, 4401 Penn Avenue, Pittsburgh, PA 15224, USA.
authors.forename: K
authors.initials: K
authors.lastname: Skvorak
authors.affiliation: Genomics and Proteomics Core Laboratories, Bioinformatics Core, University of Pittsburgh, 3343 Forbes Avenue, Pittsburgh, PA 15260, USA.
authors.forename: T
authors.initials: T
authors.lastname: Sultana
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D019175:DNA Methylation; D004195:Disease Models, Animal; D005260:Female; D015870:Gene Expression; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000502:physiology; D017042:Phenylketonuria, Maternal / Q000235:genetics*; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D017422:Sequence Analysis, DNA
chemical_list: D035683:MicroRNAs
keywords: Gene expression; Maternal PKU Syndrome; Methylome; Toxicity