Project description:To gain insight into the molecular regulation of human heart development, a detailed comparison of the mRNA and miRNA transcriptomes across differentiating human-induced pluripotent stem cell (hiPSC)–derived cardiomyocytes and biopsies from fetal, adult, and hypertensive human hearts was performed. Gene ontology analysis of the mRNA expression levels of the hiPSCs differentiating into cardiomyocytes revealed 3 distinct groups of genes: pluripotent specific, transitional cardiac specification, and mature cardiomyocyte specific. Hierarchical clustering of the mRNA data revealed that the transcriptome of hiPSC cardiomyocytes largely stabilizes 20 days after initiation of differentiation. Nevertheless, analysis of cells continuously cultured for 120 days indicated that the cardiomyocytes continued to mature toward a more adult-like gene expression pattern. Analysis of cardiomyocyte-specific miRNAs (miR-1, miR-133a/b, and miR-208a/b) revealed a miRNA pattern indicative of stem cell to cardiomyocyte specification. A biostatistitical approach integrated the miRNA and mRNA expression profiles revealing a cardiomyocyte differentiation miRNA network and identified putative mRNAs targeted by multiple miRNAs. Together, these data reveal the miRNA network in human heart development and support the notion that overlapping miRNA networks re-enforce transcriptional control during developmental specification. miRNA expression profiling of differentiating human-induced pluripotent stem cell (hiPSC)–derived cardiomyocytes (days 0-120)
Project description:Induced pluripotent stem cells (iPSCs) outwardly appear to be indistinguishable from embryonic stem cells (ESCs). A study of gene expression profiles of mouse and human ESCs and iPSCs suggests that, while iPSCs are quite similar to their embryonic counterparts, a recurrent gene expression signature appears in iPSCs regardless of their origin or the method by which they were generated. Upon extended culture, hiPSCs adopt a gene expression profile more similar to hESCs; however, they still retain a gene expression signature unique from hESCs that extends to miRNA expression. Genome-wide data suggested that the iPSC signature gene expression differences are due to differential promoter binding by the reprogramming factors. High-resolution array profiling demonstrated that there is no common specific subkaryotypic alteration that is required for reprogramming and that reprogramming does not lead to genomic instability. Together, these data suggest that iPSCs should be considered a unique subtype of pluripotent cell. Detailed analysis comparing induced pluripotent stem cells revealed functional gene expression differences to hESCs that are changed with long term passaging. We used microarrays to detail the changes that are made throughout passaging to hiPSC lines.
Project description:Chemical warfare nerve agents (CWNA) are potent cholinesterase inhibitors that may also have non-cholinesterase effects. Several in vivo studies have shown that exposure to CWNA compounds induces damage in the brain and heart. Underlying mechanisms of this damage are a critical area of research for the development of medical countermeasures. This study utilized microRNA (miRNA) analysis to evaluate potential direct cellular effects of the nerve agent VX (o-ethyl-s-[2 (diisopropylamino) ethyl] methylphosphonothiolate) on human-induced pluripotent stem cell (iPSC)-derived neurons iPSC-derived neurons were treated with VX at concentrations of 0µM (saline control), 0.1µM or 100µM for either 1 hour or 6 hours. Total RNA was then isolated and processed for miRNA microarray analysis using Affymetrix miRNA 2.0 GeneChips
Project description:Chemical warfare nerve agents (CWNA) are potent cholinesterase inhibitors that may also have non-cholinesterase effects. Several in vivo studies have shown that exposure to CWNA compounds induces damage in the brain and heart. Underlying mechanisms of this damage are a critical area of research for the development of medical countermeasures. This study utilized microRNA (miRNA) analysis to evaluate potential direct cellular effects of the nerve agent GD/soman (O-Pinacolyl methylphosphonofluoridate) on human-induced pluripotent stem cell (iPSC)-derived neurons iPSC-derived neurons were treated with GD at concentrations of 0µM (saline control), 0.1µM or 100µM for either 1 hour or 6 hours. Total RNA was then isolated and processed for miRNA microarray analysis using Affymetrix miRNA 2.0 GeneChips
Project description:Chemical warfare nerve agents (CWNA) are potent cholinesterase inhibitors that may also have non-cholinesterase effects. Several in vivo studies have shown that exposure to CWNA compounds induces damage in the brain and heart. Underlying mechanisms of this damage are a critical area of research for the development of medical countermeasures. This study utilized microRNA (miRNA) analysis to evaluate potential direct cellular effects of the nerve agent GD/soman (O-Pinacolyl methylphosphonofluoridate) on human-induced pluripotent stem cell (iPSC)-derived cardiomyocytes iPSC-derived cardiomyocytes were treated with GD at concentrations of 0µM (saline control), 0.1µM or 100µM for either 1 hour or 6 hours. Total RNA was then isolated and processed for miRNA microarray analysis using Affymetrix miRNA 2.0 GeneChips
Project description:Chemical warfare nerve agents (CWNA) are potent cholinesterase inhibitors that may also have non-cholinesterase effects. Several in vivo studies have shown that exposure to CWNA compounds induces damage in the brain and heart. Underlying mechanisms of this damage are a critical area of research for the development of medical countermeasures. This study utilized microRNA (miRNA) analysis to evaluate potential direct cellular effects of the nerve agent VX (o-ethyl-s-[2 (diisopropylamino) ethyl] methylphosphonothiolate) on human-induced pluripotent stem cell (iPSC)-derived neurons
Project description:Chemical warfare nerve agents (CWNA) are potent cholinesterase inhibitors that may also have non-cholinesterase effects. Several in vivo studies have shown that exposure to CWNA compounds induces damage in the brain and heart. Underlying mechanisms of this damage are a critical area of research for the development of medical countermeasures. This study utilized microRNA (miRNA) analysis to evaluate potential direct cellular effects of the nerve agent GD/soman (O-Pinacolyl methylphosphonofluoridate) on human-induced pluripotent stem cell (iPSC)-derived neurons