Project description:This study aimed to provide new compelling clues in the PM2.5-induced toxicity and mechanism which order the meaningful bioinformatics evidences for further study on human cardiovascular system. We used microarrays to detail the global program of gene expression underlying cellularisation and identified distinct classes of up-regulated genes after treated with chemical substance.
Project description:This study aimed to provide new compelling clues in the PM2.5-induced toxicity and mechanism which order the meaningful bioinformatics evidences for further study on human cardiovascular system. We used microarrays to detail the global programme of gene expression underlying cellularization and identified distinct classes of up-regulated genes after treated with chemical substance
Project description:Polysomes of untreated (NT) or tunicamycin-treated (TM) human cardiomyocyte AC16 cells were immunoprecipitated (IP MRPS15) with anti-MRPS15 antibody, followed by RNA sequencing. As a control, RNAseq was performed on polysome-associated RNAs of untreated or tunicamycin-treated human cardiomyocyte AC16 cells before immunoprecipitation (input).
Project description:Cardiovascular diseases are associated with an altered cardiomyocyte metabolism. Due to a shortage of human heart tissue, experimental studies mostly rely on alternative approaches including animal and cell culture models. Since the use of isolated primary cardiomyocytes is limited, immortalized cardiomyocyte cell lines may represent a useful tool as they closely mimic human cardiomyocytes. This study is focused on the AC16 cell line generated from adult human ventricular cardiomyocytes. Despite an increasing number of articles employing AC16 cells, the comprehensive proteomic, bioenergetic and oxygen-sensing characterization of proliferating versus differentiated cells is still lacking. Here, we provide a comparison of these two stages, particularly emphasizing cell metabolism, mitochondrial function, and hypoxic signalling. The label-free quantitative mass spectrometry revealed a decrease in autophagy and cytoplasmic translation in differentiated AC16, confirming their phenotype. Cell differentiation led to the global increase in mitochondrial proteins (e.g. OXPHOS proteins, TFAM, VWA8, etc.) reflected by elevated mitochondrial respiration. Fatty acid oxidation proteins were increased in differentiated cells, while the expression levels of proteins associated with fatty acid synthesis were unchanged, and glycolytic proteins were decreased. There was a profound difference between proliferating and differentiated cells in their response to hypoxia and anoxia/reoxygenation. We conclude that AC16 differentiation leads to proteomic and metabolic shifts and altered cell response to oxygen deprivation. This underscores the requirement for proper selection of particular differentiation state during experimental planning.
Project description:Transcriptomic analysis of synchonized AC16 cells treated with homoharringtonine 1 µM or Rocaglamide A 100 nM for 4h (20h after synchronization) We used microarrays to detail the global programme of gene expression
Project description:There are unique stressors in the spaceflight environment. Exposure to such stressors is associated with adverse effects on astronauts' health, including increased cancer and cardiovascular disease risks. Small extracellular vesicles (sEVs, i.e., exosomes) play a vital role in intercellular communication and regulate various biological processes contributing to their role in disease pathogenesis. To assess whether spaceflight alters sEVs transcriptome profile, sEVs were isolated from the blood plasma of 3 astronauts at two different time points: 10 days before launch (L-10) and 3 days after return (R+3) from the Shuttle mission. Human adult ventricular cardiomyocyte cells (AC16) were treated with L-10 and R+3 astronauts-derived exosomes for 24 hours. Total RNA was isolated and analyzed for gene expression profiling using Affymetrix microarrays. Enrichment analysis was performed using Enrichr. Transcription factor enrichment analysis using the ENCODE/ChEA Consensus TF database identified gene sets related to the polycomb repressive complex 2 (PRC2) and Vitamin D receptor (VDR) in AC16 cells treated with R+3 compared to cells treated with L-10 astronauts-derived exosomes. Further analysis of the histone modifications using datasets from the Roadmap Epigenomics Project confirmed enrichment in gene sets related to the H3K27me3 repressive mark. Interestingly, analysis of previously published H3K27me3–chromatin immunoprecipitation sequencing (ChIP-Seq) ENCODE datasets showed enrichment of H3K27me3 in the VDR promoter. Collectively, our results suggest that astronaut-derived sEVs may epigenetically repress the expression of the VDR in human adult cardiomyocytes c by promoting the activation of the PRC2 complex and H3K27me3 levels.
Project description:Transcriptomic analysis of synchonized AC16 cells after 0-6-12-18 or 24 h of digoxin treatment (5µM) We used microarrays to detail the global programme of gene expression