Cytosine Methylation Dysregulation in Neonates Following Intrauterine Growth Restriction

Department of Obstetrics, Gynecology and Women's Health, Albert Einstein College of Medicine, Bronx, New York, United States of America.
PLoS ONE (Impact Factor: 3.23). 01/2010; 5(1):e8887. DOI: 10.1371/journal.pone.0008887
Source: PubMed


Perturbations of the intrauterine environment can affect fetal development during critical periods of plasticity, and can increase susceptibility to a number of age-related diseases (e.g., type 2 diabetes mellitus; T2DM), manifesting as late as decades later. We hypothesized that this biological memory is mediated by permanent alterations of the epigenome in stem cell populations, and focused our studies specifically on DNA methylation in CD34+ hematopoietic stem and progenitor cells from cord blood from neonates with intrauterine growth restriction (IUGR) and control subjects.
Our epigenomic assays utilized a two-stage design involving genome-wide discovery followed by quantitative, single-locus validation. We found that changes in cytosine methylation occur in response to IUGR of moderate degree and involving a restricted number of loci. We also identify specific loci that are targeted for dysregulation of DNA methylation, in particular the hepatocyte nuclear factor 4alpha (HNF4A) gene, a well-known diabetes candidate gene not previously associated with growth restriction in utero, and other loci encoding HNF4A-interacting proteins.
Our results give insights into the potential contribution of epigenomic dysregulation in mediating the long-term consequences of IUGR, and demonstrate the value of this approach to studies of the fetal origin of adult disease.

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Available from: Reid F Thompson, Nov 11, 2014
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    • "In mice, hypermethylation in as few as one paternally expressed imprinted gene is sufficient to induce FGR (Murphy et al. 2001, McMinn et al. 2006, Dilworth et al. 2010). Additionally, global methylation changes and altered gene expression in non-imprinted genes have also been identified in FGR placentas (McCarthy et al. 2007, Einstein et al. 2010, Struwe et al. 2010). "
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    ABSTRACT: Fetal growth restriction (FGR) is a major obstetric complication stemming from poor placental development. Previously, we have shown that paternal obesity in mice is associated with impaired embryo development and significantly reduced fetal and placental weights. We hypothesised that the FGR observed in our rodent model of paternal diet-induced obesity is associated with alterations in metabolic, cell signalling and stress pathways. Male C57Bl/6 mice were fed either a normal or high fat diet for 10 weeks prior to sperm collection for IVF and subsequent embryo transfer. On embryonic day 14, placentas were collected and RNA extracted from both male and female placentas to assess mRNA expression of 24 target genes using custom RT-qPCR arrays. Peroxisome proliferator activated receptor alpha (Ppara) and caspase-12 (Casp12) expression were significantly altered in male placentas from obese fathers compared to normal (p<0.05), but not female placentas. PPARA and CASP12 protein was localised within the placenta to trophoblast giant cells by immunohistochemistry, and relative protein abundance was determined by Western Blot analysis. DNA was also extracted from the same placentas to determine methylation status. Global DNA methylation was significantly increased in female placentas from obese fathers compared to normal (p<0.05), but not male placentas. Here we demonstrate for the first time that paternal obesity is associated with changes in gene expression and methylation status of extraembryonic tissue in a sex-specific manner. These findings reinforce the negative consequences of paternal obesity prior to conception, and emphasise the need for more lifestyle advice for prospective fathers.
    Reproduction (Cambridge, England) 02/2015; 149(5). DOI:10.1530/REP-14-0676 · 3.17 Impact Factor
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    • "However, the molecular mechanisms underlying the phenomenon of fetal programming remained largely unknown. In recent years, an increasing number of studies identified epigenetic alterations at certain loci to be involved in this process of programming and adaptation [2-5]. "
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    ABSTRACT: Background and study design: Prenatal growth restriction and low birth weight have been linked to long-term alterations of health, presumably via adaptive modifications of the epigenome. Recent studies indicate a plasticity of the 11p15 epigenotype in response to environmental changes during early stages of human development. We analyzed methylation levels at different 11p15 loci in 20 growth-discordant monozygotic twin pairs. Intrauterine development was discordant due to severe twin-to-twin transfusion syndrome (TTTS), which was treated by fetoscopic laser coagulation of communicating vessels before 25 weeks of gestation. Methylation levels at age 4 were determined in blood and buccal cell-derived DNA by the single nucleotide primer extension reaction ion pair reverse-phase high performance liquid chromatography (SNuPE IP RP HPLC) assay. Methylation at LINE-1 repeats was analyzed as an estimate of global methylation.In general, variance of locus-specific methylation levels appeared to be higher in buccal cell- as compared to blood cell-derived DNA samples. Paired analyses within the twin pairs revealed significant differences at only one CpG site (IGF2 dmr0 SN3 (blood), +1.9% in donors; P = 0.013). When plotting the twin pair-discordance in birth weight against the degree of discordance in site-specific methylation at age 4, only a few CpGs were found to interact (one CpG site each at IGF2dmr0 in blood/saliva DNA, one CpG at LINE-1 repeats in saliva DNA), with 26 to 36% of the intra-twin pair divergence at these sites explained by prenatal growth discordance. However, across the entire cohort of 40 children, site-specific methylation did not correlate with SD-scores for weight or length at birth. Insulin-like growth factor-II serum concentrations showed significant within-twin pair correlations at birth (R = 0.57) and at age 4 (R = 0.79), but did not differ between donors and recipients. They also did not correlate with the analyzed 11p15 methylation parameters. In a cohort of 20 growth-discordant monozygotic twin pairs, severe alteration in placental blood supply due to TTTS appears to leave only weak, if any, epigenetic marks at the analyzed CpG sites at 11p15.
    Clinical Epigenetics 03/2014; 6(1):6. DOI:10.1186/1868-7083-6-6 · 4.54 Impact Factor
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    • "A higher angle formed when HpaII count is high relative to MspI is evidence of relative hypomethylation while a smaller angle is evidence of hypermethylation19. Prior studies in our lab have demonstrated a correlation of ~0.9 between replicates and between angle values obtained by HELP-tagging and quantification of methylation by bisulphite MassArray (Sequenom)192829. "
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    ABSTRACT: While DNA methylation plays a role in T-helper (Th) cell maturation, its potential dysregulation in the non-atopic Th1-polarized systemic inflammation observed in obesity-associated asthma is unknown. We studied DNA methylation epigenome-wide in peripheral blood mononuclear cells (PBMCs) from 8 obese asthmatic pre-adolescent children and compared it to methylation in PBMCs from 8 children with asthma alone, obesity alone and healthy controls. Differentially methylated loci implicated certain biologically relevant molecules and pathways. PBMCs from obese asthmatic children had distinctive DNA methylation patterns, with decreased promoter methylation of CCL5, IL2RA and TBX21, genes encoding proteins linked with Th1 polarization, and increased promoter methylation of FCER2, a low-affinity receptor for IgE, and of TGFB1, inhibitor of Th cell activation. T-cell signaling and macrophage activation were the two primary pathways that were selectively hypomethylated in obese asthmatics. These findings suggest that dysregulated DNA methylation is associated with non-atopic inflammation observed in pediatric obesity-associated asthma.
    Scientific Reports 07/2013; 3:2164. DOI:10.1038/srep02164 · 5.58 Impact Factor
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