Project description:EZH2 plays an important role in stem cell renewal and maintenance by inducing gene silencing via its histone methyltransferase activity. EZH2 downregulation markedly enhances neuron differentiation of human mesenchymal stem cells (hMSCs)chromatin at promoters of EZH2 target genes. comparison of knockdown EZH2 of hMSCs vs hMSCs
Project description:EZH2 plays an important role in stem cell renewal and maintenance by inducing gene silencing via its histone methyltransferase activity. Previously, we showed EZH2 downregulation markedly enhances neuron differentiation of human mesenchymal stem cells (hMSCs). To understand how EZH2 regulates neuron differentiation of hMSCs, we wanted to identify the target genes of EZH2. For this reasons we performed ChIP-on-chip experiments using specific EZH2 antibodies followed by a human promoter array for the whole human genome.
Project description:EZH2 plays an important role in stem cell renewal and maintenance by inducing gene silencing via its histone methyltransferase activity. Previously, we showed EZH2 downregulation markedly enhances neuron differentiation of human mesenchymal stem cells (hMSCs). To understand how EZH2 regulates neuron differentiation of hMSCs, we wanted to identify the target genes of EZH2. For this reasons we performed ChIP-on-chip experiments using specific EZH2 antibodies followed by a human promoter array for the whole human genome. The 3A6-hMSCs were differentiated into neuron for 5 days, and then 109 cells were harvested for the ChIP-on-chip assay. The procedure was based on the manufacturer's instructions (NimbleGen).
Project description:EZH2 plays an important role in stem cell renewal and maintenance by inducing gene silencing via its histone methyltransferase activity. EZH2 downregulation markedly enhances neuron differentiation of human mesenchymal stem cells (hMSCs)chromatin at promoters of EZH2 target genes.
Project description:In order to study the role of the HDAC9 in human mesenchymal stem cell differentiation, gene expression analysis was performed with inducible silencing of HDAC9 in human mesenchymal stem cell purchasing from Cyagen Biosciences (HUXMA-90011, Guangzhou, China). Transcriptomic analysis performed on mRNA of human mesenchymal stem cells transfected with lentiviral knockdown HDAC9 (lenti-HDAC9) particles revealed down and up regulation of transcripts of hMSCs differentiation genes
Project description:Cell-fate determination of human mesenchymal stem/stromal cells (hMSCs) is precisely regulated by lineage-specific transcription factors and epigenetic enzymes. We found that CTR9, a key scaffold subunit of Polymerase Associated Factor Complex (PAFc), selectively regulates hMSC differentiation to osteoblasts and chondrocytes, but not to adipocytes. An in vivo ectopic osteogenesis assay confirmed the essentiality of CTR9 in hMSC-derived bone formation. CTR9 counteracts the activity of EZH2, the epigenetic enzyme that deposits H3K27me3, in hMSCs. Accordingly, CTR9 knockdown (1) hMSCs gain H3K27me3 mark, and the osteogenic differentiation defects of CTR9 KD hMSCs can be partially rescued by treatment with EZH2 inhibitors. Transcriptome analyses identified Bone Morphology Protein-2 (BMP-2) as a downstream effector of CTR9. BMP-2 secretion, membrane anchorage, as well as the BMP-SMAD pathway were impaired in CTR9 KD MSCs, and the effects were rescued by BMP-2 supplementation. This study uncovers an epigenetic mechanism engaging CTR9-H3K27me3-BMP-2 axis to regulate osteochondral lineage differentiation of hMSCs. To investigate the function of CTR9 in the gene regulation of early osteogenic committment , we established human MSCs in which CTR9 gene has been knocked down by two individual shRNAs (shControl vs shCTR9#3/#5).
Project description:To determine the miRNAs potentially responsible for the premature-senescence mediated gene expression regulation in human adipose-derived mesenchymal stem cells, we performed illumine bead microarray analyses on parental hMSCs, and globally depleted miRNAs by inhibiting DGCR8, an essential component of miRNA biogenesis. DGCR8 knockdown in hMSCs resulted in severe proliferation defects with senescence-associated changes including markedly increased reactive oxygen species (ROS) levels. In gene expression profiling, thousand two hundred and forty seven transcripts were upregulated and 1,136 genes were downregulated in Dgcr8 knockdown cells compared to control cells (p < 0.05, Fisher’s exact test).
Project description:Analysis of serum starved prelamin A-accumulating hMSCs at gene expression level. The hypothesis tested in the present study was that prelamin A accumulation induces the dysregulation of genes that are essensial for cell survival under a stress condition such as serum starvation. The results provide important information about these genes and the functional categories that are dysregulated due to prelamin A accumulation in serum starved hMSCs. Two samples are analyzed in this microarray experiment: human mesenchymal stem cell cultured under serum starvation conditions (during 24 hours) which accumulate prelamin A (pre-hMSCs) and control mesenchymal stem cells (ctrl-hMSCs). 2 biological replicates (hMSCs derived from 2 different bone marrow donors) and 1 technical replicate are included in this analysis.