Journal: Scientific Reports

Article Title: Prenatal maternal stress and wheeze in children: novel insights into epigenetic regulation

doi: 10.1038/srep28616

Figure Lengend Snippet: ( A ) Pie charts represent the number and percentage of DMRs observed in mothers and children. Two classes of DMRs were differentiated: gDMRs for which methylation significantly correlated with a meQTL-SNP and ngDMRs where the underlying sequence variation did not influence methylation (red: hypermethylation, blue: hypomethylation). ( B ) The table summarizes the functional genomic regions enriched for an overlap with these DMRs. Active regulatory genomic regions in particular enhancer elements were significantly enriched in overlapping DMRs irrespective of their type (gDMR or ngDMR).

Article Snippet: Methylation differences of selected differentially methylated regions (DMRs) were assessed using the MassARRAY system (Sequenom Inc./Agena Bioscience GmbH, Hamburg Germany), a mass spectrometry based approach for targeted methylation analysis .

Techniques: Methylation, Sequencing, Functional Assay

Journal: Scientific Reports

Article Title: Prenatal maternal stress and wheeze in children: novel insights into epigenetic regulation

doi: 10.1038/srep28616

Figure Lengend Snippet: Pathways intertwined by stress induced differential methylation.

Article Snippet: Methylation differences of selected differentially methylated regions (DMRs) were assessed using the MassARRAY system (Sequenom Inc./Agena Bioscience GmbH, Hamburg Germany), a mass spectrometry based approach for targeted methylation analysis .

Techniques: Methylation

Journal: Scientific Reports

Article Title: Prenatal maternal stress and wheeze in children: novel insights into epigenetic regulation

doi: 10.1038/srep28616

Figure Lengend Snippet: ( A ) Based on the histone modification data the DMR (WGBS: Δmeth = 28%) identified in the NMUR1 gene is located in an enhancer region (ENCODE/Active Elements (own ChIP-Seq data)). ( B ) Validation of the NMUR1 DMR (7 CpGs, chr2: 232,394,701–232,394,805) in the total cohort by MassARRAY shows that an increase of the maternal stress level leads to a significant elevation in the methylation level in children at time of birth. In highly stressed children this methylation increase corresponds to a significantly decreased mRNA expression of NMUR1 (mean +/− 95% CI, whiskers +/− non-outlier range). Relationship between NMUR1 methylation and maternal stress score were determined by multiple regression analysis adjusted for gender of the child, birth week, age of the mother, mode of delivery, maternal smoking/-medication during pregnancy, parental history of atopy and cell composition. ( C ) Relationship of interleukin-4, -5 and -6 secretion at time of birth and NMUR1 methylation. Data are presented as ratios of the mean (MR) and 95% confidence intervals. Models were adjusted for known confounders of interleukin concentrations in cord blood (month of birth, mode of delivery, gender of the child, parental history of atopy, smoking during pregnancy and cell composition).

Article Snippet: Methylation differences of selected differentially methylated regions (DMRs) were assessed using the MassARRAY system (Sequenom Inc./Agena Bioscience GmbH, Hamburg Germany), a mass spectrometry based approach for targeted methylation analysis .

Techniques: Modification, ChIP-sequencing, Biomarker Discovery, Methylation, Expressing

Journal: Scientific Reports

Article Title: Prenatal maternal stress and wheeze in children: novel insights into epigenetic regulation

doi: 10.1038/srep28616

Figure Lengend Snippet: ( A ) Although found in an intron the DMR (WGBS: Δmeth = 22%) in the GNA11 gene is located in an enhancer region according to our own ChIP-Seq data. ChIA-PET data suggest this enhancer regulates its host gene. ( B ) Maternal stress and the methylation level in the GNA11 DMR show a significant relationship at time of birth while at the same time a significant reverse relationship is observed for the expression of GNA11 when including all children in our cohort for whom persistent or never persistent wheeze was reported (mean +/− 95% CI, whiskers +/− non-outlier range, p-values adjusted for: gender of the child, birth week, age of the mother, mode of delivery, maternal smoking/-medication during pregnancy, parental history of atopy and cell composition). ( C ) In four-year-old children increased methylation in the GNA11 enhancer (chr19: 3110675) is associated with an increased risk for persistent wheeze. Corresponding results are observed for GNA11 expression. Odds ratios were calculated using logistic regression models adjusted for known confounders of wheezing (parental history of atopy, parental educational level, gender, siblings, smoking during pregnancy, ETS exposure after birth, cat keeping and maternal stress score).

Article Snippet: Methylation differences of selected differentially methylated regions (DMRs) were assessed using the MassARRAY system (Sequenom Inc./Agena Bioscience GmbH, Hamburg Germany), a mass spectrometry based approach for targeted methylation analysis .

Techniques: ChIP-sequencing, ChIA Pet Assay, Methylation, Expressing

Journal: Scientific Reports

Article Title: Prenatal maternal stress and wheeze in children: novel insights into epigenetic regulation

doi: 10.1038/srep28616

Figure Lengend Snippet: ( A ) Methylation could only be assessed for one CpG in the CACNB4 ngDMR. Amplification of the entire region yielded no PCR product therefore we could only consider two CpGs (chr2: 152898920–152899112). Since one of the two CpGs was a CpG destroying SNP further analysis was restricted to a single position. ( A ) At time of birth methylation of this CpG shows a significant relationship to the maternal stress score (considering all children either persistently wheezing or showing no respiratory symptoms), ( B ) while in the same children expression of CACNB4 is decreased to maternal stress. Adjusted p-values were calculated including gender of the child, birth week, age of the mother, mode of delivery, maternal smoking/-medication during pregnancy, parental history of atopy and cell composition as confounding variables.

Article Snippet: Methylation differences of selected differentially methylated regions (DMRs) were assessed using the MassARRAY system (Sequenom Inc./Agena Bioscience GmbH, Hamburg Germany), a mass spectrometry based approach for targeted methylation analysis .

Techniques: Methylation, Amplification, Expressing