Expression data from mouse pancreatic cell lines treated with chromatin-targeting small molecules
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ABSTRACT: We measured the genome-wide expression changes induced by 29 compounds targeting HDACs, DNMTs, histone lysine methyltransferases (HKMTs), and protein arginine methyltransferases (PRMTs) in pancreatic M-NM-1- and M-NM-2-cell lines. We used microarrays to detail the global programme of gene-expression changes induced by chromatin-targeting compounds at 1, 6, and 24 hours. To identify a comprehensive list of target genes that can be regulated by modulating the activities of chromatin-modifying enzymes, we measured the genome-wide transcriptional effects of 29 compounds in pancreatic alpha and beta cell lines at three time points. The results indicate that compounds cause similar effects independent of the cell line in which they were profiled. All clinical HDAC inhibitors fell into the structural classes of hydroxamic acids and ortho-amino anilides, respectively and up- and down-regulated hundreds of transcripts. In contrast, more selective compounds like the HKMT inhibitor BIX-01294 have specific effects.
Project description:Expression of insulin in terminally differentiated non-beta pancreatic cell types could be important for treating type-1 diabetes. We observed that the kinase inhibitor GW8510 up-regulated insulin expression in mouse pancreatic alpha cells. Gene-expression profiling and gene-set enrichment analysis of GW8510-treated alpha cells suggested coordinate up-regulation of the p53 pathway, which was further implicated in induction of insulin transcript. In order to determine a potential mechanism of insulin induction by GW8510, we treated mouse alpha cells (aTC1) and beta cells (bTC3) with 3.3 µM GW8510 or 0.1% DMSO for five days, and performed gene-expression profiling followed by gene-set enrichment analysis (GSEA). Three biological replicates were included per condition. GW8510 treatment in each cell line was compared to DMSO-treated control.
Project description:We measured the genome-wide expression changes induced by 29 compounds targeting HDACs, DNMTs, histone lysine methyltransferases (HKMTs), and protein arginine methyltransferases (PRMTs) in pancreatic α- and β-cell lines. We used microarrays to detail the global programme of gene-expression changes induced by chromatin-targeting compounds at 1, 6, and 24 hours.
Project description:The lack of accurate in vitro assays for predicting in vivo toxicity of chemicals together with new legislations demanding replacement and reduction of animal testing has triggered the development of alternative methods. This study aimed at developing a transcriptomics-based in vitro prediction assay for in vivo genotoxicity. The transcriptomics changes induced in the human liver cell line HepG2 by 34 compounds after treatment for 12h, 24h and 48h were used for the selection of gene-sets that can discriminate between in vivo genotoxins (GTX) and in vivo non-genotoxins (NGTX). By combining publicly available results for these chemicals from standard in vitro genotoxicity studies with transcriptomics, we developed several prediction models. These models were validated by means of an additional set of 28 chemicals. The study investigated differential gene expression in HepG2 cell line mRNA following 12 hours of exposure to 34 different compounds and their solvents; 24 and 48 hours of exposure to 62 different compounds and their solvents. Three biological replicates per compound/solvent. In total 560 arrays .
Project description:Cytokine-induced beta-cell apoptosis is a key event for the death of pancreatic beta cells in the development of type-1 diabetes. We identified BRD0476 as a novel suppressor of cytokine-induced beta-cell apoptosis. We used microarrays to look for gene set(s) that are regulated by BRD0476. Rat INS-1E cells were treated with cytokine cocktails (IL-1b, IFN-g and TNF-a) and/or BRD0476 for 6 or 12 hours. Total RNAs were isolated using the RNEasy kit from Qiagen.
Project description:Microarray analysis of HT-29 cells co-cultured with tumor necrosis factor (TNF-a) in the presence or absence of polymeric formula as used for Exclusive Enteral Nutrition (EEN) therapy. Results provide insights into the molecular mechanisms underlying the anti-inflammatory effect of polymeric formula on intestinal epithelium. Total RNA obtained from 9 samples of HT-29 cells. Six samples were treated with TNF-a in the presence (3 samples) or absence (3 samples) of Polymeric Formula. Three samples were untreated and used as a control.
Project description:Copy number and Gene expression profiling of HT-29 wild-type and bortezomib resistant cell lines Identification of mechanisms of bortezomib resistance Gene expression differences between HT-29 cell line variants
Project description:The homeodomain transcription factor, Pdx-1, has important roles in pancreatic development and M-NM-2-cell function and survival. In the present study, we demonstrate that adenovirus-mediated overexpression of Pdx-1 in rat or human islets also stimulates cell replication. Moreover, co-overexpression of Pdx-1 with another homeodomain transcription factor, Nkx6.1, has an additive effect on proliferation compared to either factor alone, implying discrete activating mechanisms. Consistent with this, Nkx6.1 stimulates mainly M-NM-2-cell proliferation, whereas Pdx-1 stimulates both M-NM-1- and M-NM-2-cell proliferation. Furthermore, cyclins D1/D2 are upregulated by Pdx-1 but not by Nkx6.1, and inhibition of cdk4 blocks Pdx-1- but not Nkx6.1-stimulated islet cell proliferation. Genes regulated by Pdx-1 and not Nkx6.1 were identified by microarray analysis. Two members of the transient receptor potential cation (TRPC) channel family, TRPC3 and TRPC6, are upregulated by Pdx-1 overexpression, and siRNA-mediated knockdown of TRPC3/6 or TRPC6 alone inhibits Pdx-1-induced but not Nkx6.1-induced islet cell proliferation. Pdx-1 also stimulates ERK1/2 phosphorylation, an effect partially blocked by knockdown of TRPC3/6, and blockade of ERK1/2 activation with a MEK1/2 inhibitor partially impairs Pdx-1-stimulated proliferation. These studies define a pathway by which overexpression of Pdx-1 activates islet cell proliferation that is distinct from and additive to a pathway activated by Nkx6.1. We identified genes that were upregulated or downregulated at 48 h with Pdx-1 overexpression as compared to untreated and M-NM-2gal controls. We set up a microarray using primary rat islets that were left untreated or transduced with adenoviruses overexpressing M-NM-2gal or Pdx-1 for 48 h.
Project description:The goal of this study was to identify transcriptional changes induced by a novel compound BRD9876 as a starting point to identify the compound's mechanism of action. Compound-treated cells are compared in duplicate with cells treated vehicle control (DMSO).
Project description:The transcriptomics changes induced in the human liver cell line HepG2 by 17 hepatotoxic compounds, 5 non-hepatotoxic compounds and solvent controls after treatment for 24h The study investigated differential gene expression in HepG2 cell line mRNA following 24 hours of exposure to 17 hepatotoxic compounds, 5 non-hepatotoxic compounds and solvent controls. Three biological replicates per compound/solvent. In total 105 arrays .