ChIP-Seq for cofactors in cancer
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Richard Young
PROVIDER: E-GEOD-45984 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Similar Datasets
Project description:Discovery of the genome-wide location of proteins using ChIP-Seq has allowed global mapping of the key transcription factors and chromatin regulators that control gene expression programs in various cells. Many DNA-associated processes are targeted for disease therapy. This study investigates the functions of small molecule therapeutics that target DNA-associated processes involved of CDK9 and BRD4. Genomic DNA was enriched by chromatin immunoprecipitation (ChIP) and analyzed by Solexa sequencing. ChIP was performed using an antibody against RNAP2, BRD4, CDK9, and CTCF using whole cell extract (WCE) as a background control.
2013-04-23 | E-GEOD-43743 | biostudies-arrayexpress
Project description:An ability to map the global interactions of a chemical entity with chromatin genome-wide could provide new insights into the mechanisms by which a small molecule perturbs cellular functions. we developed a method that uses chemical derivatives and massively parallel DNA sequencing (Chem-Seq) to identify the sites bound by small chemical molecules throughout the human genome. We developed in vivo and in vitro Chem-Seq protocols with a biotinylated derivative of small molecules. In the in vivo protocol, Cells were first treated with biotinylated ligand and cross-linked with formaldehyde at the same time. Cells were then lysed, sonicated to shear the DNA, and streptavidin beads were used to isolate biotinylated ligand and associated chromatin fragments. We then used massively parallel sequencing to identify the enriched DNA fragments, and mapped these sequences to the genome. In in vitrol protocol, MM1.S cells were fixed and the derived sonicated lysate incubated with biotinylated drug to enrich for bound chromatin regions in vitro. We then used massively parallel sequencing to identify the enriched DNA fragments, and mapped these sequences to the genome.
2013-05-31 | E-GEOD-44098 | biostudies-arrayexpress
Project description:Chromatin regulators have become highly attractive targets for cancer therapy, yet many of these regulators are expressed in a broad range of healthy cells and contribute generally to gene expression. An important conundrum has thus emerged: how can inhibition of a general regulator of gene expression produce selective effects at specific oncogenes? Here we investigate how inhibition of the transcriptional coactivator BRD4 (Bromodomain containing 4) leads to selective inhibition of disease-critical oncogenes in a highly malignant blood cancer, multiple myeloma (MM). We found that BRD4 generally occupies the promoter elements of active genes together with the Mediator coactivator, but remarkably high levels of these two coactivator proteins were associated with a small set of exceptionally large enhancers. These super-enhancers are associated with genes that feature prominently in MM biology, including the MYC oncogene. Treatment of MM tumor cells with the BET-bromodomain inhibitor JQ1 led to preferential loss of BRD4 at super-enhancers and consequent transcription elongation defects that preferentially impact genes with super-enhancers, including the c-MYC oncogene. Super-enhancers were found at key oncogenic drivers in many other tumor cells. Thus, super-enhancers can regulate oncogenic drivers in tumor cells, which in some cells can be preferentially disrupted by BRD4 inhibition, which in turn contributes to the selective transcriptional effects observed at these oncogenes. These observations have implications for the discovery of novel cancer therapeutics directed at components of super-enhancers in diverse tumor types. ChIP-Seq for chromatin regulators and RNA Polymerase II in multiple myeloma, glioblastoma multiforme, and small cell lung cancer
2013-01-02 | E-GEOD-42355 | biostudies-arrayexpress
Project description:P493-6 cells are immortalized human peripheral B cells that carry a conditional, tetracycline-regulated myc gene. We present ChIP-seq analysis of key transcritional regulators in P493-6 cells expressing various levels of c-Myc: 0hr (low c-Myc levels), 1hr (intermediate c-Myc levels), 24hr (very high c-Myc levels) and No Tet (steady-state c-Myc levels). Brd4, c-Myc, Max, Med1, RNAPII, and the chromatin modification H3K27Ac were profiled in P493-6 cells
2012-11-14 | E-GEOD-42262 | biostudies-arrayexpress
Project description:Discovery of the genome-wide location of proteins using ChIP-Seq has allowed global mapping of the key transcription factors and chromatin regulators that control gene expression programs in various cells. Many DNA-associated processes are targeted for disease therapy. This study investigates the functions of small molecule therapeutics that target DNA-associated processes involved with CDK9 and BRD4. Genomic DNA was enriched by chromatin immunoprecipitation (ChIP) and analyzed by Solexa sequencing. ChIP was performed using an antibody against Brd4, RNA Polymerase II, Med1, H3K27ac, and CDK8 as well as whole-cell extract (WCE) DNA controls
2013-12-24 | E-GEOD-53601 | biostudies-arrayexpress
Project description:MM1.S cells are an aggressive dexamethasone sensitive multiple myeloma cell line whose transcritional program is driven by deregulated c-Myc activity. We present ChIP-seq analysis of key transcritional regulators that are implicated the c-Myc transcriptional network in MM1.S cells treated with vehicle or 500nM JQ1. Brd4, Cdk9, cMyc, Max, Med1, RNA Pol II, and the chromatin modifications H3K4me3 and H3K27Ac were profiled in MM1.S cells treated with 500nM JQ1 for 24hr
2012-11-30 | E-GEOD-42161 | biostudies-arrayexpress
Project description:The bromodomain and extraterminal (BET) protein Brd4 is a validated drug target in leukemia, yet its regulatory function in this disease is not well understood. Here, we show that Brd4 chromatin occupancy in acute myeloid leukemia closely correlates with the hematopoietic transcription factors (TFs) Pu.1, Fli1, Erg, C/EBPα, C/EBPβ, and Myb at nucleosome-depleted enhancer and promoter regions. We provide evidence that these TFs, in conjunction with the lysine acetyltransferase activity of p300/CBP, facilitate Brd4 recruitment to their occupied sites to promote transcriptional activation. Moreover, chemical inhibition of BET bromodomains is found to suppress the functional output each hematopoietic TF, thereby interfering with essential lineage-specific transcriptional circuits in this disease. These findings reveal a chromatin-based signaling cascade comprised of hematopoietic TFs, p300/CBP, and Brd4, which supports leukemia maintenance and is suppressed by BET bromodomain inhibition. ChIP-Seq for regulatory factors of Brd4 in MLL-AF9 transformed acute myeloid leukemia cells (RN2)
2015-05-01 | E-GEOD-66122 | biostudies-arrayexpress
Project description:To enable successful SLC function deorphanization we aimed to select a minimal number of cell lines covering expression of as many SLCs as possible. Based on a publicly available RNA-Seq dataset of 675 cell lines (Klijn C. et al, Nat. Biotechnol. 2015) we selected a set of 6 adherent human cell lines (HCT 116, Huh 7, LS180, MDA MB 468, SK MEL 28, 1321N1) cumulatively covering expression (TPM>1) of about 80% of all human SLCs. These cell lines will be used to generate SLC-knockout and SLC-overexpressing single cell clones. Additionally, Jump-In T-REx HEK293 cells will be used for generating SLC-overexpression cell lines. In order to characterize the expressed proteome of these parental cell lines, we performed extensive proteomic profiling using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Mass spectra were acquired on a Q Exactive hybrid quadrupole-Orbitrap mass spectrometer (Thermo Fisher Scientific) in data-dependent acquisition (DDA) mode. Instrument performance was monitored by a quality control routine using BSA- and HeLa digest standards. Data was analyzed using Thermo Proteome Discoverer 2.3 software. This data release consist of (1) RAW mass spectrometry data files generated by the instrument per fraction, (2) ProteomeDiscoverer result files containing all Peptide-Spectrum-Matches (PSMs), peptide and protein grouping, scoring and confidence values, as well as abundance values from label-free-quantitation (LFQ). As RESOLUTE will generate a significant number of protein-protein interaction data sets (AP-MS and BioID), the presented whole proteome profiling data set can serve as a reference core proteome data set, providing I) technical information on detected and expressed tryptic peptides and proteins; and II) provide valuable information on quantitative distribution of i.e. bait and prey proteins in the different cell lines. In conjunction with the other Omics data sets, like the previously published transcriptomics data, the characterization of the proteome will complement and enable valuable insight into the biological state of the investigated cell lines under default culturing conditions. Furthermore, members of the research community might apply advanced analyses with a focus out of the scope of the RESOLUTE project (e.g. PTM analysis).
2024-04-29 | PXD051747 | Pride
Project description:The bromodomain and extraterminal (BET) protein Brd4 is a validated drug target in leukemia, yet its regulatory function in this disease is not well understood. Here, we show that Brd4 chromatin occupancy in acute myeloid leukemia closely correlates with the hematopoietic transcription factors (TFs) Pu.1, Fli1, Erg, C/EBPα, C/EBPβ, and Myb at nucleosome-depleted enhancer and promoter regions. We provide evidence that these TFs, in conjunction with the lysine acetyltransferase activity of p300/CBP, facilitate Brd4 recruitment to their occupied sites to promote transcriptional activation. Moreover, chemical inhibition of BET bromodomains is found to suppress the functional output each hematopoietic TF, thereby interfering with essential lineage-specific transcriptional circuits in this disease. These findings reveal a chromatin-based signaling cascade comprised of hematopoietic TFs, p300/CBP, and Brd4, which supports leukemia maintenance and is suppressed by BET bromodomain inhibition. PolyA selected RNA-Seq for drug treated or shRNA-expressing MLL-AF9 transformed acute myeloid leukemia cells (RN2)
2015-05-01 | E-GEOD-66068 | biostudies-arrayexpress
Project description:BRCA2 maintains genome stability by facilitating DNA repair via homologous recombination and replication fork stability. Loss of BRCA2 is deleterious for survival of normal cells, but is paradoxically tolerated in cancer cells. Using quantitative mass-spectrometry, differences in protein expression were identified that might shed light on how breast cancer cells (HCC38) survive in the absence of BRCA2.
2018-12-19 | PXD007253 | Pride