Micro-RNA signatures in monozygotic twins discordant for congenital heart defects

Abstract

BackgroundMicroRNAs (miRNAs) are small RNAs regulating gene expression post-transcriptionally. Recent studies demonstrated that miRNAs are involved in the development of congenital heart defects (CHD). In this study, we aimed at identifying the specific patterns of miRNAs in blood of monozygotic twin pairs discordant for CHD and to assess whether miRNAs might be involved in the development or reflect the consequences of CHD.MethodsmiRNA microarray analysis and Real-Time Quantitative PCR (RT-qPCR) were employed to determine the miRNA abundance level from 12 monozygotic twins discordant for CHD and their non-CHD co-twins (n = 12). Enrichment analyses of altered miRNAs were performed using bioinformatics tools.ResultsCompared with non-CHD co-twins, profiling analysis indicated 34 miRNAs with a significant difference in abundance level (p<0.05, fold change ≥ 1.3), of which 11 miRNAs were up-regulated and 23 miRNAs were down-regulated. Seven miRNAs were validated with RT-qPCR including miR-511-3p, miR-1306-5p, miR-421, miR-4707-3p, miR-4732-3p, miR-5189-3p, and miR-890, and the results were consistent with microarray analysis. Five miRNAs namely miR-511-3p, miR-1306-5p, miR-4732-3p, miR-5189-3p, and miR-890 were found to be significantly up-regulated in twins < 10 years old. Bioinformatics analysis showed that the 7 validated miRNAs were involved in phosphatidylinositol signaling, gap junction signaling, and adrenergic signaling in cardiomyocytes.ConclusionsOur data show deregulated miRNA abundance levels in the peripheral blood of monozygotic twins discordant for CHD, and identify new candidates for further analysis, which may contribute to understanding the development of CHD in the future. Bioinformatics analysis indicated that the target genes of these miRNAs are likely involved in signaling and communication of cardiomyocytes.

MicroRNA-29b/c-3p Indicate Advanced Liver Fibrosis/Cirrhosis in Univentricular Heart Patients With and Without Fontan Palliation

“…DataAssist TM Software v3.0 (Applied Biosystems) was used to calculate the relative abundance changes of miRNAs by the equation 2 − Ct with RNAU6B serving as an endogenous control as previously used and validated for this type of sample (21,23,24,(26)(27)(28)(29)(30)(31). Adjustment for multiple testing was performed by controlling the false discovery rate (FDR) according to the approach of Benjamini and Hochberg (32).…”

“…These platforms contain probes for the detection of 2,549 human miRNAs. An input amount of 100 ng of isolated RNA including miRNAs was labeled and subsequently hybridized to the miRNA microarray chip and the procedure was completed as previously described ( 23 ).…”

Integrated microRNA and mRNA Expression Profiling Identifies Novel Targets and Networks Associated with Ebstein’s Anomaly

“…After loading, the array was placed in the Biomark HD instrument for quantification and detection using a specific thermal cycling protocol for either miRNA or mRNA analysis. RNAU6B and GAPDH were used as reference endogenous controls for miRNA and mRNA, respectively, as previously recommended for this type of sample [ 14 , 15 , 16 , 17 , 18 , 19 , 21 , 26 ]. The data were analyzed with Real-Time PCR Analysis Software (Fluidigm Corporation), according to Fluidigm’s recommendation.…”

“…Currently, there are 2300 “real” miRNAs in the miRNA database based on miRBase.org [ 13 ]. It is assumed, that almost every cellular and biological process is regulated by miRNAs, including processes regulating CHDs [ 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 ].…”

Global DNA Methylation as a Potential Underlying Mechanism of Congenital Disease Development

“…Study showed that the MTHFR genotype/diet combination and BMI were significantly associated with LINE-1 methylation in mothers of children with DS/ CHD + [5]. Recently, micro-RNA signatures discordant for CHD in monozygotic twins were observed [112].…”