Project description:The human LncRNA microarray analysis of the 6 monocytes samples from Coronary Artery Disease patients and non Coronary Artery Disease 3 Coronary Artery Disease patients and 3 non-Coronary Artery Disease donors
Project description:The human LncRNA microarray analysis of the 6 monocytes samples from Coronary Artery Disease patients and non Coronary Artery Disease
Project description:LncRNA expression profiling for 6 human monocytes samples from Coronary Artery Disease patients and non Coronary Artery Disease patients
Project description:The human LncRNA microarray analysis of the 6 plasma samples from Coronary Artery Disease patients and non Coronary Artery Disease Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images. Quantile normalization was performed using Expander6 and subsequent data processing was performed using the GeneSpring GX v11.5.1 software package (Agilent Technologies). After low intensity filtering, LncRNAs and mRNAs that at least 2 out of 12 samples have flags in Present or Marginal (“All Targets Value”) were chosen for quantile normalization and further data analysis. Differentially expressed LncRNAs and mRNAs with statistical significance were identified through Volcano Plot filtering. Pathway analysis and GO analysis were applied to determine the roles of these differentially expressed mRNAs played in these biological pathways or GO terms. Finally, Hierarchical Clustering was performed to show the distinguishable LncRNAs expression pattern among samples.
Project description:This study aimed to develop a short-read RNA-Seq protocol to detect mRNAs, lncRNAs and circRNAs (including putative novel lncRNAs) in human plasma. This protocol was applied to plasma from patients with established stable CAD (samples collected ~4 months after an acute coronary event) and healthy controls to screen for candidate mRNA, lncRNA and circRNA biomarkers associated with the presence of coronary artery disease and the progression from CAD to HF. We hypothesised that circulating levels of non-coding RNAs in patients with stable CAD may relect progression of atherosclerotic disease or adverse myocardial remodeling, and may help identify patients at risk of future cardiovascular events. Here we demonstrate that mRNAs, lncRNAs and circRNAs can be reliably detected in human plasma by deep RNA-Seq and report novel candidate biomarkers for the presence of CAD.
Project description:In this research, two 4-hydroxy-2-nonenal modified peptides with differential performance were identified in the plasma of patients with coronary artery disease, whether their antibodies were different in the plasma of patients with coronary artery disease and healthy people.
Project description:In this study, multiomics profiling, including CUT&Tag, promoter capture Hi-C (PCHi-C), and microarray, identified LncRNA-ITGA2 as a novel elncRNA that was highly expressed in PDGF-induced proliferative human VSMCs. Notably, LncRNA-ITGA2 was significantly increased in both plasma and coronary atherosclerotic tissues of patients with coronary artery disease (CAD) compared with control subjects. Loss- and gain-of-function studies revealed that LncRNA-ITGA2 potently increased PDGF-induced VSMC proliferation and migration. Moreover, LncRNA-ITGA2 overexpression enhanced neointimal hyperplasia in vivo in a mouse carotid artery injury model, exhibiting its partial functional conservation. RNA-sequencing and CRISPR-Cas9 gene-editing technology revealed integrin α2 (ITGA2) as a downstream target of LncRNA-ITGA2 in VSMCs. Mechanistically, promoter-enhancer interactions were detected by PCHi-C at the ITGA2 locus after PDGF-BB treatment. Chromatin immunoprecipitation sequencing (ChIP-Seq) and chromatin isolation by RNA purification (ChIRP)-qPCR showed that LncRNA-ITGA2 directly bound to the Enhancer-ITGA2 and increased H3K27 acetylation in both the Enhancer-ITGA2 and Promoter-ITGA2. In addition, we demonstrated, by ChIRP-MS and RNA immunoprecipitation, that LncRNA-ITGA2 interacted with the DNA binding-protein NONO (non-pou domain containing octamer-binding protein), which also bound to the ITGA2 promoter, as verified by the ChIP-qPCR assay. Ultimately, the knockout cell lines of NONO, LncRNA-ITGA2, and its promoter confirmed the above regulatory mechanism.
Project description:We have employed whole genome microarray expression profiling as a discovery platform to identify genes involved in plaque rupture. Human coronary artery endothelial cells (ECs) were stimulated in vitro for 12 hours with plasma obtained from the coronary sinus (CS) and the aorta (Ao) of patients with ACS (n=8), or patients with stable angina (SA, n=4). For each patient, gene expression profile was evaluated by microarray technology in ECs exposed to plasma obtained from CS and compared to that of ECs exposed to plasma sampled from Ao. In patient with ACS we found 684 genes up-regulated and 283 down-regulated was observed as compared to patients with SA. Functional and network analyses of statistically significant gene showed that the up regulated genes were associated to pathways IL17 Signaling. To validate the microarray data the RNAs were used for Real Time PCR experiment. Human coronary artery endothelial cells (ECs) were stimulated in vitro for 12 hours with plasma obtained from the coronary sinus (CS) and the aorta (Ao) of patients with ACS (n=8), or patients with stable angina (SA, n=4). For each patient, gene expression profile was evaluated by microarray technology in ECs exposed to plasma obtained from CS and compared to that of ECs exposed to plasma sampled from Ao.
