Project description:Neonatal sepsis is an important health-care concern worldwide occurring more frequently in premature newborns and for which diagnosis is yet a challenge, causing delays in therapy and increasing the risk of death. DNA methylation, involved in regulating gene expression, has been associated with the development and progression of sepsis. Actually, the detection of differentially methylated regions (DMRs) is a promising epigenetic tool used for diagnosis and prognosis in complex diseases. The present study focuses on two different bioinformatic methods, DMRcate and mCSEA, with the aim of obtaining different methylation traits using Illumina Infinium Human Methylation EPIC data of neonatal sepsis patients. The DMR sets obtained by both approaches can also be overlapped to obtain a reliable set of DMRs which can contribute to improve our understanding on the molecular pathways underlying disease.
Project description:Genome wide DNA methylation differences in umbilical cord blood (UCB) of preterm and term born neonates. The Illumina 450K Human DNA methylation Beadchip was used to obtain the DNA methylation profiles of 12 preterm and 12 term UCB samples across 450,000 CpGs. 2 samples were removed due to failed quality control, and BMIQ normalization was done with 11 preterm and 11 term samples.
Project description:Endothelial colony-forming cells were isolated and expanded from the mononuclear cell fraction (MNC) obtained from the cord blood of term (CT) and preterm (PT) neonates, and characterized as previously described (Vassallo PF et al 2014). The objective of the study was to identify differentially expressed genes between CT and PT neonates. ECFC were harvested from cultures dishes at passage 3 and total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion), according to the manufacturerâs recommendations. Cy3-CTP labeled RNA was prepared according to standard Agilent protocol from 400ng total RNA. The hybridization was performed for 17 hrs at 65°C. cRNA labeling and hybridization performance were performed and all parameters checked were found within the manufacturers specifications. Arrays were scanned as described in the manufacturersâ protocol. Signal intensities on 20 bit tiff images were calculated by Feature Extraction software (FE, Version 8.5; Agilent Technologies). Data analyses were conducted with GeneSpring GX software (Vers.13.1.1; Agilent Technologies)
Project description:Bronchopulmonary dysplasia (BPD) is the major cause of chronic lung disease (CLD) and morbidity in preterm infants, which characterized by impaired pulmonary alveolar development in preterm infants. There is increasing evidence that micro RNA (miRNA) has a close association with the development of pulmonary diseasesincluding lung growth, pulmonary fibrosis, pneumonia, etc. The potential role of miRNA in the pathogenesis of BPD is unclear. This study aims to clarify the role of adrenomedullin (ADM) regulated by miRNA-547-3p during the process of BPD and illustrate the protective effect of ADM involved in preterm infants.We indentified the differential expressed miRNA and their potential target genes, microarray analysis using Agilent Human 4x44K Gene Expression Microarrays v2 and miRCURY LNA™ microRNA Array was performed. The results of miRNA chip were scanned by Axon GenePix 4000B and the signal of probes were read by GenePix Pro 6.0 software. Only the target genes which exist in both two databases and having different expression trend were remained. Thus, miRNA-574-3p and it’s target gene ADM were selected to do the further research in our study. And we verified miRNA-574-3p and ADM expression in BPD premature infants by realtime q-PCR. After the microarray expression analysis, there were 516 probes differential expressed between BPD samples and normal samples. Among them, 510 were up-regulated and 213 were down-regulated. Meanwhile, to the miRNA aspect, there were 37 up-regulated and 44 down-regulated. After text mining, we found ADM was already found play a role in BPD. ADM is an endogenous peptide with potent angiogenic, anti-oxidant, and anti-inflammatory properties. Realtime q-PCR assay results showed that, miRNA-574-3p expression level in blood samples from preterm infants with BPD was significantly decreased, ADM expression level in blood samples from preterm infants with BPD was significantly increased. In our study, we found that up-regulation of ADM regulated by miRNA-574-3p could protect preterm infants with BPD. These data provide novel insights into ADM regulated by miRNA-574-3p which may be shed light on BPD prevention and treatment.
Project description:Bronchopulmonary dysplasia (BPD) is the major cause of chronic lung disease (CLD) and morbidity in preterm infants, which characterized by impaired pulmonary alveolar development in preterm infants. There is increasing evidence that micro RNA (miRNA) has a close association with the development of pulmonary diseasesincluding lung growth, pulmonary fibrosis, pneumonia, etc. The potential role of miRNA in the pathogenesis of BPD is unclear. This study aims to clarify the role of adrenomedullin (ADM) regulated by miRNA-547-3p during the process of BPD and illustrate the protective effect of ADM involved in preterm infants.We indentified the differential expressed miRNA and their potential target genes, microarray analysis using Agilent Human 4x44K Gene Expression Microarrays v2 and miRCURY LNA™ microRNA Array was performed. The results of miRNA chip were scanned by Axon GenePix 4000B and the signal of probes were read by GenePix Pro 6.0 software. Only the target genes which exist in both two databases and having different expression trend were remained. Thus, miRNA-574-3p and it’s target gene ADM were selected to do the further research in our study. And we verified miRNA-574-3p and ADM expression in BPD premature infants by realtime q-PCR. After the microarray expression analysis, there were 516 probes differential expressed between BPD samples and normal samples. Among them, 510 were up-regulated and 213 were down-regulated. Meanwhile, to the miRNA aspect, there were 37 up-regulated and 44 down-regulated. After text mining, we found ADM was already found play a role in BPD. ADM is an endogenous peptide with potent angiogenic, anti-oxidant, and anti-inflammatory properties. Realtime q-PCR assay results showed that, miRNA-574-3p expression level in blood samples from preterm infants with BPD was significantly decreased, ADM expression level in blood samples from preterm infants with BPD was significantly increased. In our study, we found that up-regulation of ADM regulated by miRNA-574-3p could protect preterm infants with BPD. These data provide novel insights into ADM regulated by miRNA-574-3p which may be shed light on BPD prevention and treatment.
Project description:Bronchopulmonary dysplasia (BPD) is a lung disease in premature infants characterized by impaired pulmonary development which persists into later life. While advances in neonatal care have improved survival rates of premature infants, cases of BPD haves been increased. Therapeutic options are limited for prevention and treatment. This study was designed to explore the relationship between gestational age (GA), birth weight and estímate blood cell-type composition in premature infants and to elucidate early epigenetic biomarkers associated with BPD. Peripheral blood DNA (at days 14 and 28) from preterm neonates that went on to develop BPD (n = 14) or not (nonBPD, n = 93) was applied to Illumina EPIC methylation arrays. Using DNA methylation analysis of cord blood DNA, we investigated association of GA and birth weight with the estimated distribution of cord blood cell types, particularly the nucleated red blood cell (NRBC) in a pilot-size cohort of preterm infants with or without BPD. We describe changes in methylation-based estimates of blood cell-type composition in relation to GA and birth weight. After adjusting for covariates (GA, birth weight, cell type proportions, etc.) we identify differentially methylated CpGs and genes associated with BPD at different time points.