Transcriptional effects of exogenous lipid on differentiated THP-1 cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Dr Laura Sadofsky
PROVIDER: E-MTAB-4153 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:There is currently no human macrophage cell line and in vitro experiments requiring these cells currently require mitogenic stimulation of a macrophage precursor cell line (for example THP-1 cells) or ex vivo maturation of circulating primary monocytes. We have derived a unique sub-clone of the THP-1 cell line capable of spontaneous and perpetual differentiation into alveolar macrophage-like cells. Here we used the Agilent G4851C (v3) array to look at the gene expression of the new cell line which can be compared it with THP-1 cells and PMA stimulate THP-1 cells (Accession number E-MTAB-4153). In addition the new human alveolar macrophage like cell line has been treated with unmodified lipid to look at lipid uptake by these cells and the effect on gene expression.
2020-01-31 | E-MTAB-7169 | biostudies-arrayexpress
Project description:Genome-wide comparisons of transcription factor binding sites in different species allow for a direct evaluation of the evolutionary constraints that shape transcription factor binding landscapes. To gain insights into the evolution of the PPARg-dependent transcriptional network we obtained binding data for PPARg, RXR and PU.1 in human macrophages and compared the profiles to matching data from mouse macrophages. We found that PPARg binding was highly divergent and only 5% of the PPARg bound regions were occupied in both species. Despite the low conservation of PPARg binding sites, conserved PPARg target genes contribute more than 30% to the functional target genes identified in human macrophages. In addition conserved target genes are strongly enriched for lipid metabolic functions. We detected the lineage-specification factor PU.1 at the majority of human PPARg binding sites. This confirmed the juxtaposed binding configuration found in mouse macrophages and demonstrated the preservation of tissue-specific adjacent PPARg-Pu.1 binding in the absence of individual binding site conservation. Finally, based on this of PPARg and PU.1 binding between human and mouse we suggest a mechanism by which PU.1 facilitates PPARg binding site turnover in macrophages. Comparison of RNAs level in Rosiglitazone (RSG) and vehicle (DMSO) treated PMA differentiated THP-1 cells over a timecourse from 0.5h to 12h
2011-12-01 | E-GEOD-25137 | biostudies-arrayexpress
Project description:We analysed the capacity of THP-1 cells (differentiated to macrophagoid cells) to recognize RNA sequences via pattern recognition receptors in vitro. Gene expression was analysed by RNA-Microarray. Cytokine production was analysed by ELISA assays. We used microarrays to investigate differential gene expression in THP-1 cell line undifferentiated in comparison with 3 days or 8 days differentiated with phorbol myristate acetate (PMA). Microarray analysis revealed differential gene expression patterns of THP-1 when differentiated. THP-1 cells, undifferentiated, 3 days PMA-differentiated and 8 days PMA-differentiated
2015-11-09 | E-GEOD-67264 | biostudies-arrayexpress
Project description:Macrophages play a key role in both innate and adaptive immunity, but our knowledge on the changes in transcription regulation that occurs during their differentiation from monocytes is still limited. In this study, we used a meta-analysis followed by a systems biology approach for the identification of differentially expressed genes between monocytes and macrophages and possible regulators of these changes in transcription. Based on the pattern of gene expression change, transcription regulator analysis predicted a decrease in Enhancer of Zeste homolog 2 (EZH2), a histone 3 lysine 27 methyl transferase, activity after differentiation of monocytes into macrophages. This inhibition was validated by a significant decrease in trimethylated H3K27 during differentiation of both human primary monocytes into macrophages and the THP-1 cell line into macrophage-like cells. Overexpressing EZH2 during differentiation of monocytes and THP-1 cells obstructs cellular adhesion, thus preventing the first step in differentiation. Another facet of macrophage differentiation is the cessation of proliferation, and inhibition of EZH2 by the small molecule inhibitor GSK126 in THP-1 cells indeed impedes proliferation. This study shows an important part for epigenetic changes during monocyte differentiation. It highlights the role of EZH2 activity behind the changes needed in adhesion and proliferation mechanisms for macrophage formation. THP-1s were differentiated into macrophage like cells by PMA stimulation.
2013-12-28 | E-GEOD-53691 | biostudies-arrayexpress
Project description:Previous reports have shown low vitamin D serum levels and polymorphisms in the vitamin D receptor (VDR) to be associated with increased risk for TB. Given that 1?,25-dihydroxyvitamin D3 has a role in lipid metabolism control, we tested whether the link between 1?,25-dihydroxyvitamin D3 and tuberculosis involves macrophage lipid metabolism. Since formation of lipid droplets (LD) is a hallmark of lipid dysregulation in M. tuberculosis-infected macrophages, we measured LD content as a readout of altered lipid metabolism in infected THP-1 cells. Induction of LD, which peaked by 24 hours post-infection was prevented by addition of 1?,25-dihydroxyvitamin D3 at the time of infection. To investigate the mechanism of 1?,25-dihydroxyvitamin D3 modulation of LD formation, we analyzed the transcriptome of M. tuberculosis-infected THP-1 cells with and without 1?,25-dihydroxyvitamin D3 treatment. THP-1 cells were cultured in 24-well flat bottom plates, pre-treated with 20ng/ml PMA for 24 h, and incubated with M. tuberculosis H37Rv. After 4 hours of infection, cells were washed to remove extracellular bacteria and treated with 100 nM of 1?,25-dihydroxyvitamin D3. THP-1 cells were collected and analyzed at 24h. 3 biological replicates in each of the 4 groups were prepared: Infected, Infected plus 1?,25-dihydroxyvitamin D3, Non-infected, Non-infected plus 1?,25-dihydroxyvitamin D3.
2014-07-01 | E-GEOD-57028 | biostudies-arrayexpress
Project description:Analysis of interferon-stimulated genes (ISGs) in various primary cells and immortalized cell lines, following type 1 interferon (IFN) treatment. Some cell types become resistant to HIV-1 infection following type 1 interferon treatment (such as macrophages, THP-1, PMA-THP-1, U87-MG cells and to a lesser extent, primary CD4+ T cells) while others either become only partially resistant (e.g., HT1080, PMA-U937) or remain permissive (e.g., CEM, CEM-SS, Jurkat T cell lines and U937); for more information see (Goujon and Malim, Journal of Virology 2010) and (Goujon and Schaller et al., Retrovirology 2013). We hypothesized that the anti-HIV-1 ISGs are differentially induced and expressed in restrictive cells compared to permissive cells and performed a whole genome analysis following type 1 IFN treatment in cell types exhibiting different HIV-1 resistance phenotypes. 48 samples; design: 9 cell lines, primary CD4+ T cells and primary macrophages, untreated and IFN-treated; 2 replicate experiments per cell line; 3 replicate experiments per primary cell type
2013-09-13 | E-GEOD-46599 | biostudies-arrayexpress
Project description:The data is supplementary to the RNA-seq analysis of LPS and palmitate stimulation of THP-1 macrophages (E-MTAB-6064), where palmitate for the cell treatment was dissolved in sodium hydroxide and coupled with BSA at a molar ratio 7.5:1. Here we stimulated THP-1 macrophages with the corresponding concentration of BSA (4%) and NaOH (0.4 mM) in the presence of 10 nM phorbol 12-myristate 13-acetate (PMA) for 24 hours added to the standard RPMI 1640 medium with 10% fetal bovine serum (FBS) to estimate the effects and compared them with unstimulated THP-1 macrophages cultured in RPMI 1640 medium with 10% FBS and 10nM PMA.
2019-05-17 | E-MTAB-6439 | biostudies-arrayexpress
Project description:We report a whole-genome expression profiling of PMA-treated THP-1 human macrophages as a host system, infected by W-Beijing strains of different sublineages or the H37Rv reference strain. Surprisingly, we found that host transcriptional responses were irrespective of inter-sublineage variations. Based on this finding, we further showed that such common host transcriptional responses were probably under the control of a regulatory network involving STATs, IRF-1, IRF-7 and Oct-1 transcriptional factors. We also suggested that these putative regulators might act in a cooperative manner to induce the expression of host-responsive genes, particularly those involved in cytokine-cytokine receptor interactions. THP-1 cells were infected by 11 different sublineages plus one reference strain H37Rv (two replications) at three time points (i.e., 4h, 18h and 48h). For each infection as well as a non-infection control, RNA was extracted using Trizol (Life Technologies)TRIZOL® reagent (Invitrogen), and subsequently amplified and labeled with biotin according to the standard Affymetrix® protocol. Specifically, RNA was DNase-treated and purified with RNeasy columns (Qiagen). The purified RNA was quantified using the ND-1000 Spectrophotometer (NanoDrop Technologies) and Agilent 2100 Bioanalyzer (Agilent Technologies). The biotinylated cRNA was then fragmented and subjected to hybridization on Human Genome U133 Plus 2.0 Array (Affymetrix, USA). The scanned images were converted to cell intensity files (CEL) using GeneChip® Operating Software (GCOS) (Affymetrix). These CEL raw expression data were normalized using Robust Multi-array Averaging (RMA) with quantile normalization in R (Bioconductor). Detection call-based filter was applied to remove all the probe-sets whose expression values were consistently below an empirically-determined value of minimum sensitivity, which were evaluated according to the 95th percentile of all the ‘Absent’ call-flagged signals of the entire dataset. Following the normalization and filtering, a gene expression matrix (termed as TB expression matrix) was constructed. The TB expression matrix was then subjected to component plane presentation integrated self-organizing map (CPP-SOM) for visual comparisons, or subjected to Cluster3.0/TreeView-1.0.8 softwares with Euclidean distance algorithm for sample classification of 13 time-course transcriptome profiles. Also, Linear Models for Microarray Data (LIMMA) bioconductor library[18] was applied to identify those differentially expressed genes between any two successive time points. The criteria for identifying the top significant genes for the designed contrast was based on P-values (< 0.01) corrected using Benjamini and Hochberg false discovery rate (FDR) procedure. Sets of genes of interest such as differentially expressed genes between 4h and 18h infections were uploaded onto Database for Annotation, Visualization and Integrated Discovery (DAVID) v6.7 for identifying enriched gene ontology and KEGG pathways (FDR<0.01). For TFBS enrichment analysis, we applied PRIMA (PRomoter Integration in Microarray Analysis) program to identify TFs whose binding sites are significantly over-represented in a given gene set (i.e., differentially expressed genes) compared to background. Bonferroni-corrected P-values (< 0.01) were utilized to determine the significance of the binding site enrichments of TFs. Such computationally predicted TFs were considered as candidate regulators responsible for the putative co-regulations of differentially expressed genes.
2011-12-01 | E-GEOD-29628 | biostudies-arrayexpress
Project description:RNA-Seq was carried out in order to obtain the expression profile of lipopolysaccharide (LPS)-induced transcriptome changes in PMA-differentiated human THP-1 cell line.
2021-10-17 | E-MTAB-10513 | biostudies-arrayexpress
Project description:R. conorii (pathogenic) and R. montanensis (non-pathogenic) display opposite survival versus death phenotypes in macrophage-like cells, respectively. We herein employed a global transcriptomic profiling of host responses to infection (1hpi) of human THP-1 macrophages with R. conorii and R. montanensis. Shortly, total RNA was harvested from uninfected, R. conorii- and R. montanensis-infected THP-1 macrophages, DNA was removed from RNA purification (DNAse treatment), ribosomal RNA was depleted and cDNA libraries were constructed. The samples were sequenced using an Ion Proton V2 chip on Ion Chef Instrument. The programmes Cufflinks and Cuffmerge were used to map transcripts and calculate gene expression, and Cuffdiff was used to calculate which samples had genes, which were statistically significantly differentially expressed between conditions.
2019-12-01 | E-MTAB-6724 | biostudies-arrayexpress