title: Epigenetic Regulation of Placenta-Specific 8 Contributes to Altered Function of Endothelial Colony-Forming Cells Exposed to Intrauterine Gestational Diabetes Mellitus.
abstract: Intrauterine exposure to gestational diabetes mellitus (GDM) is linked to development of hypertension, obesity, and type 2 diabetes in children. Our previous studies determined that endothelial colony-forming cells (ECFCs) from neonates exposed to GDM exhibit impaired function. The current goals were to identify aberrantly expressed genes that contribute to impaired function of GDM-exposed ECFCs and to evaluate for evidence of altered epigenetic regulation of gene expression. Genome-wide mRNA expression analysis was conducted on ECFCs from control and GDM pregnancies. Candidate genes were validated by quantitative RT-PCR and Western blotting. Bisulfite sequencing evaluated DNA methylation of placenta-specific 8 (PLAC8). Proliferation and senescence assays of ECFCs transfected with siRNA to knockdown PLAC8 were performed to determine functional impact. Thirty-eight genes were differentially expressed between control and GDM-exposed ECFCs. PLAC8 was highly expressed in GDM-exposed ECFCs, and PLAC8 expression correlated with maternal hyperglycemia. Methylation status of 17 CpG sites in PLAC8 negatively correlated with mRNA expression. Knockdown of PLAC8 in GDM-exposed ECFCs improved proliferation and senescence defects. This study provides strong evidence in neonatal endothelial progenitor cells that GDM exposure in utero leads to altered gene expression and DNA methylation, suggesting the possibility of altered epigenetic regulation.
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Emily K
authors.initials: EK
authors.lastname: Blue
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: BreAnn M
authors.initials: BM
authors.lastname: Sheehan
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Zia V
authors.initials: ZV
authors.lastname: Nuss
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Frances A
authors.initials: FA
authors.lastname: Boyle
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Caleb M
authors.initials: CM
authors.lastname: Hocutt
authors.affiliation: Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Cassandra R
authors.initials: CR
authors.lastname: Gohn
authors.affiliation: Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Kaela M
authors.initials: KM
authors.lastname: Varberg
authors.affiliation: Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, IN.
authors.forename: Jeanette N
authors.initials: JN
authors.lastname: McClintick
authors.affiliation: Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN Department of Cellular and Integrative Physiology, Indiana University School of Medicine, Indianapolis, IN Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN Indiana University Melvin and Bren Simon Cancer Center, Indiana University School of Medicine, Indianapolis, IN lhanelin@iu.edu.
authors.forename: Laura S
authors.initials: LS
authors.lastname: Haneline
mesh_terms: D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence; D018899:CpG Islands; D019175:DNA Methylation; D016640:Diabetes, Gestational / Q000503:physiopathology*; D042783:Endothelial Cells / Q000502:physiology*; D044127:Epigenesis, Genetic; D005260:Female; D006801:Humans; D052250:Nitric Oxide Synthase Type III / Q000235:genetics; D011247:Pregnancy; D011506:Proteins / Q000235:genetics*; D013234:Stem Cells / Q000502:physiology*; D014599:Uterus / Q000378:metabolism*
chemical_list: C506004:PLAC8 protein, human; D011506:Proteins; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III