Project description:Coronary artery disease (CAD) poses a worldwide health threat. Compelling evidence shows that pericardial adipose tissue (PAT), a brown-like adipose adjacent to the external surface of the pericardium, is associated with CAD. However, the specific molecular mechanisms of PAT in CAD are elusive. For characterizing human PAT and explore its association with CAD, the transcriptome characteristics were assessed in 5 CAD patients and 4 controls via RNA-sequencing.
Project description:affy_rnai_cadpoplar - affy_rnai_cadpoplars - This experiment aims to characterize global gene expression in young xylem of transgenic RNAi-CAD poplars in comparison to WT poplars. Cinnamyl Alcohol Dehydrogenase (CAD) is the final enzyme involved in the monolignol biosynthesis pathway. Transgenic poplars were produced using RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003) and exhibited the expected phenotype (red xylem, reduced CAD activity).Biological question (15 lines max):This experiment aims to characterize global gene expression in young xylem of transgenic RNAi-CAD poplars in comparison to WT poplars. Cinnamyl AlcoholDehydrogenase (CAD) is the final enzyme involved in the monolignol biosynthesispathway.Transgenic poplars were produced using RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003) and exhibited the expectedphenotype (red xylem, reduced CAD activity). -RNAi-CAD transgenic poplars were produced using hairpin RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003). For this transcriptome anaylsis, 2 independent transgenic lines (named pHG8-CAD2 and pHG8-CAD19) from the same transformation procedure were used as biological repeats. Four-month-old poplar plants were inclined at 30° in the greenhouse and sampled after 26 days. Young differentiating xylem originating from the lower side of stems - opposite wood - (ODX) was sampled on each individual tree by scrapping slightly the debarked stem with a scalpel. Samples were immediately flash frozen in liquid nitrogen, ground with mortar and pestle, and total RNAs were extracted from fine ground powder using the QIAGEN miRNeasy kit according to the manufacturer. One Affymetrix slide corresponds to a pool of RNA samples from 2-4 individual trees (WT, RNAi-CAD transgenic lines 2 and 19). Total number of slides = 2 genotypes (WT/RNAi line) x 1 tissue x 2 biological replicates = 4 slides were done. 4 arrays - poplar; normal vs rnai mutant comparaison
Project description:affy_rnai_cadpoplar - affy_rnai_cadpoplars - This experiment aims to characterize global gene expression in young xylem of transgenic RNAi-CAD poplars in comparison to WT poplars. Cinnamyl Alcohol Dehydrogenase (CAD) is the final enzyme involved in the monolignol biosynthesis pathway. Transgenic poplars were produced using RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003) and exhibited the expected phenotype (red xylem, reduced CAD activity).Biological question (15 lines max):This experiment aims to characterize global gene expression in young xylem of transgenic RNAi-CAD poplars in comparison to WT poplars. Cinnamyl AlcoholDehydrogenase (CAD) is the final enzyme involved in the monolignol biosynthesispathway.Transgenic poplars were produced using RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003) and exhibited the expectedphenotype (red xylem, reduced CAD activity). -RNAi-CAD transgenic poplars were produced using hairpin RNAi strategy (pHellsgate 8 vector, Helliwell and Waterhouse, 2003). For this transcriptome anaylsis, 2 independent transgenic lines (named pHG8-CAD2 and pHG8-CAD19) from the same transformation procedure were used as biological repeats. Four-month-old poplar plants were inclined at 30° in the greenhouse and sampled after 26 days. Young differentiating xylem originating from the lower side of stems - opposite wood - (ODX) was sampled on each individual tree by scrapping slightly the debarked stem with a scalpel. Samples were immediately flash frozen in liquid nitrogen, ground with mortar and pestle, and total RNAs were extracted from fine ground powder using the QIAGEN miRNeasy kit according to the manufacturer. One Affymetrix slide corresponds to a pool of RNA samples from 2-4 individual trees (WT, RNAi-CAD transgenic lines 2 and 19). Total number of slides = 2 genotypes (WT/RNAi line) x 1 tissue x 2 biological replicates = 4 slides were done.
Project description:Purpose: We applied the next-generation sequencing to explore the differential gene expression in CAD cells differentiated with dexamethasone (Dex), compared to the undifferentiated cells using Illumna -based RNA-sequencing. Methods: Total RNAs were extracted from undifferentiated CAD cells and Dex-differentiated cells (5 days). Three independent samples were collected from both experimental groups. Samples then underwent to the sequencing step performed on NextSeq with a 75 base-pair end read length. Quality filtered reads were mapped to the mouse reference genome (GRCm38.p6). Feature Counts software was implemented to obtain raw counts for all mouse genes. The gene counts were used for differential expression analysis with the DESeq2 package to identify differentially expressed genes (DEGs) in the Dex-differentiated CAD cells, as compared to the undifferentiated CAD cells. Results: The sequence reads (expression level) per sample was obtained in a range of 9.8-11.2 million. The high quality of sequence reads was mapped to the mouse reference transcriptome (GRCm38.p6). The gene counts were used for differential expression analysis. The sample-to-sample distance demonstrated that the clustering of the undifferentiated groups was completely separated from the clustering of Dex-differentiated groups. A PCA plot showed that there was a huge variation (PC 1 =99%) between the Dex-differentiated group and the undifferentiated group but less variation was observed within the same group (PC2 = 0%). These statistic profiles indicate that the transcriptomic profile between Dex-differentiated CAD cells and undifferentiated CAD cells had the different pattern. A total of 1,183 differentially expressed genes (DEGs) were identified in the Dex-differentiated CAD cells compared to the undifferentiated CAD cells (false discovery rate < 0.01 and |log2 [fold change] | > 2). Of these, 803 and 380 genes were significantly upregulated and downregulated in the Dex-differentiated group, respectively. Conclusion: Dexamethasone causes extensive changes in gene expression of CAD cells.
Project description:MicroRNAs (miRNAs) are involved in the formation of atherosclerosis. However, the alteration of miRNA profile in epicardial adipose tissue (EAT) during atherosclerosis is still uncovered. In this study, we compared the miRNA expression profiles of EAT from non-coronary atherosclerosis disease (CAD) and CAD patients, and found that 250 miRNAs were differentially expressed in CAD patients, which were associated with metabolism, ECM and inflammation process. Among the top 20 up-regulated miRNAs, we found that the expression levels of miR-200 family members (hsa-miR-200b/c-3p, miR-141-3p and miR-429), which were rich in endothelial cells, were increased in EAT from CAD patients significantly.
Project description:Expression profiling of lymphoblast cell lines of autisms from Simons Simplex Families. This experiment aims to utilize blood expression data to study the functional impact and to prioritize rare variants potentially associated with autism. Total RNA obtained from lmyphoblast cell lines derived from 439 Simons Simplex Families.