Screening for differential methylation status in fetal myocardial tissue samples with ventricular septal defects by promoter methylation microarrays

Abstract

Abstract.To identify and provide a global assessment of dna methylation in fetal ventricular septal defect (VSd), genomic dna extracted from fetal myocardial tissue samples with VSd (n=21) and from normal fetal myocardial tissue samples (n=15) was analyzed for gene methylation using array-based technology. Furthermore, the Kiaa0310, raB43, SiVa1 and ndrG2 genes were randomly selected for validation analysis using methylation-specific PCR. Our results revealed that 70 and 85 genes were regulated by hypermethylation and hypomethylation, respectively, in VSd. different clusters of genes were associated with functions including embryo development, signal transduction, cell apoptosis and cell proliferation. In conclusion, this study identified a set of candidate genes whose expression is regulated by dna methylation in fetal VSd. Introductioncongenital heart disease (cHd) is the most common type of developmental defect, occurring in almost 1% of all neonates (1). Ventricular septal defect (VSd) is the most commonly recognized cHd (2). VSd may exist alone or as an integral part of complex cHd (3). VSd is a multifactorial complex disease, in which genetic and environmental factors play important roles. despite the availability of several surgical techniques to treat VSd, the exact molecular mechanism of this type of cHd remains unclear.dna methylation has been widely recognized as a potent mechanism for silencing gene expression and maintaining genome stability (4). dna methylation is the predominant epigenetic alteration occurring in mammalian genomes, and plays a critical functional role in development, differentiation and disease (5). Previous studies have revealed that during embryonic development, the mammalian genome undergoes profound reprogramming of dna methylation patterns in the germ and early pre-implantation embryos (6). Furthermore, the different prototypes of genes in each cell, tissue and organ are thought to be regulated by dna methylation even during early development (7).The recent advent of array-based techniques offers the opportunity for more comprehensive DNA methylation profiling (8). Comparing the DNA methylation profiles of myocardial tissue samples from VSd and normal fetuses, we provide novel information for identifying gene methylation that may be implicated in the pathological consequences of VSd. Materials and methodsTissue samples. Fetal myocardial tissue samples were obtained from nanjing Maternal and child Health Hospital. Myocardial tissue samples from 21 VSd and 15 normal fetuses at 26 weeks of gestation were obtained during surgery for pregnancy termination owing to trauma of the pregnant women. all samples were collected with the approval of the appropriate institute ethics committee, and written consent was provided by each pregnant woman and her family. The speciments were immediately snap frozen in liquid nitrogen and then stored at -80˚C until analysis.DNA methylation profiling by methylated DNA immunoprecipitation. The methylation profiling by methylated DNA immunoprecipitatio...

CITED2 Mutation and methylation in children with congenital heart disease

“…Although gene mutations are potential causes for the development of CHD [3], various environmental factors that affect mother during the pregnancy also play an important role in promoting and developing of CHD [4]. For example, epigenetic modifications and gene methylation changes in fetus during pregnancy can affect gene expression levels during the development [5-7]. Aberrant methylation in the promoter region of genes can inhibit gene transcription activities by preventing the binding of transcription factor from the target genes, which is one of the potential causes of gene silence and disease [8].…”

Maternal Exposure to Nitrogen Dioxide, Intake of Methyl Nutrients, and Congenital Heart Defects in Offspring

“…Previous work exploring methylation patterns in cardiac tissue from fetuses with VSD (vs. controls) showed hypomethylation of 2 apoptosis-related genes (SIVA1 and MDM2) among only the VSD samples (46). However, there were also genes that were hypermethylated in case samples.…”

Profiling Genome-Wide DNA Methylation Patterns in Human Aortic and Mitral Valves

“…This process is heritable as well as tissue-specific and plays a major role in various physiological processes linked to cardiogenesis such as cardiomyocyte development, maturation and cardiac regeneration [9,10]. Aberrant changes in methylation profiles are associated with cardiovascular diseases (CVDs) such as ventricular septal defects, tetralogy of Fallot, atherosclerosis as well as other CVDs that lead to end-stage heart failure [11][12][13][14].…”