Transcriptional response to the HSP70 inhibitor MAL3-101 in parental rhabdomyosarcoma cells and isogenic acquired-resistance lines.
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ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE80525 | GEO | 2016/05/15
SECONDARY ACCESSION(S): PRJNA319193
REPOSITORIES: GEO
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Project description:We demonstrate that inhibition of HSP70 with MAL3-101 induces activation of the unfolded protein response, and that cells with acquired resistance upregulate a cytosolic HSP70 at the level of mRNA. Whole transcriptome sequencing was undertaken of parental or acquired-resistance RMS13 cells treated with either DMSO or 10 micromolar MAL3-101.
2016-05-15 | E-GEOD-80525 | biostudies-arrayexpress
Project description:Molecular chaperones prevent the accumulation of toxic proteins in the cell, but when chaperones are overwhelmed, the rapid modulation of proteostasis pathways provide corrective responses. One molecular chaperone in particular, Hsp70, plays a central role in both cellular protection and the response to proteotoxic insults. This is perhaps most evident in cancer cells, some of which overexpress Hsp70 and thrive even when harboring high levels of misfolded, aggregated, and/or unassembled protein complexes. In an effort to define how cells compensate for compromised proteostasis, we examined the adaptive response in breast cancer cells after challenge with a specific Hsp70 inhibitor, MAL3-101 and further define the pathways involved in the differential MAL3-101 response between sensitive and resistant cells.
2021-07-01 | GSE178352 | GEO
Project description:The major heat shock protein Hsp70 has been shown to form a complex with a scaffold protein Bag3, linking it to multiple signaling pathways. Via these interactions, the Hsp70-Bag3 module functions as a proteotoxicity sensor that controls cell signaling. Here, as a tool to identify signaling pathways regulated by this complex, we utilized JG-98, an allosteric inhibitor of Hsp70 that blocks its interaction with Bag3. Gene expression profiling followed by the pathway analysis indicated that a set of signaling pathways including the unfolded protein response (UPR) was activated by JG-98. Surprisingly, only the translation initiation factor eIF2a-associated branch of the UPR was activated under these conditions, while other UPR branches mediating induction of ER chaperones were not induced, suggesting that the response was not related to ER proteotoxicity and thus to ER-associated kinase PERK1. Indeed, induction of the UPR genes under these conditions was dependent on activation of a distinct cytoplasmic eIF2a kinase, HRI. We demonstrated that the Hsp70-Bag3 complex directly interacted with HRI and regulated phosphorylation of eIF2a upon induction of cytoplasmic proteotoxicity. Therefore, we uncovered a novel signaling response, which regulates cell death upon the buildup of abnormal protein species in cytoplasm via an Hsp70-Bag3-HRI-eIF2a axis.
2021-04-14 | GSE171440 | GEO
Project description:From preliminary experiments, HSP70 deficient MEF cells display moderate thermotolerance to a severe heatshock of 45.5 degrees after a mild preshock at 43 degrees, even in the absence of hsp70 protein. We would like to determine which genes in these cells are being activated to account for this thermotolerance. Keywords: thermal stress, heat shock response, knockout, cell culture, hsp70
2009-04-09 | GSE11120 | GEO
Project description:Parkinsonâs disease (PD) and Dementia with Lewy bodies (DLB) are the two most common examples of synucleinopathies, i.e. human disorders that display intracellular deposition of the Alpha-synuclein (aSYN) protein. The two forms of aSYN deposits, intracellular Lewy bodies and Lewy neurites, are in the PD brain primarily found in the substantia nigra whereas in DLB brains they are diffusely spread in cortical and limbic areas. We are currently analyzing the protective effects on aSYN pathology by the molecular chaperone hsp70 in mice overexpressing wildtype human aSYN. Moreover, transgenic mice overexpressing hsp70 have been cross-bred with the aSYN mouse line. Intriguingly, the aSYN / hsp70 double transgenics display a pronounced reduction in biochemically assessed aSYN aggregation. Gene expression of these mice will not only provide insight into the disease mechanisms of aSYN pathology but its prevention by the overexpression of hsp70, leading to new potential avenues of treatment. Project 1: To investigate and compare alterations in gene expression profiles of brains from wild-type mice and mice overexpressing wildtype human aSYN, human hsp70 and aSYN / hsp70, respectively. The comparison of the expression profiles of the groups of mice: aSYN, Hsp70, aSYN/ Hsp70 and non-transgenic littermates will help to identify the genes that are involved in the mediation of aggregation toxicity of aSYN expression as well as the genes that may mediate an attenuation of aSYN pathology in hsp70-coexpressors. Project 1: Selection of animals Mice of 4 month old (n=6) of age of each background (aSYN, Hsp70, aSYN/ Hsp70 and non-transgenic littermates) will be used for RNA-extraction for a total of 24 mice. Preparation of samples For the proposed experiments, we will use the recommended procedures, as outlined by the consortium, for extracting and purifying RNA from whole brain homogenate. Briefly, RNA will be extracted by the trizol method and purified by using the RNeasy MinElute Cleanup (QIAGEN Inc., USA). RNA labeling and hybridization procedures will be carried out by the microarray consortium. SPECIAL NOTE: The samples will be sent upon availability. Gene Expression Microarray Analysis Six mice will be included in each group per timepoint to generate statistically testable data and limit the possible confound of individual mouse gene expression variations. The genome expression will be evaluated through the use of the GeneChip® Mouse Genome 430 2.0 Array (Affymetrix) for each individual mouse constituting a total of 24 arrays for the proposed study. Gene expression data interpretation will be assisted by the Microarray Consortium Center bioinformatics core.
2008-06-13 | E-GEOD-4758 | biostudies-arrayexpress
Project description:The goal of this study is to assess the role of ASH1 like histone lysine methyltransferase (ASH1L) in the biology of anaplastic thyroid cancer. CRISPR-Cas9 was used to create 4 independent cell lines derived from BHT-101 anaplastic thyroid cancer cells with premature stop codons prior to the catalytic domain within both alleles of ASH1L. RNA-seq was performed on these 4 KO cell lines, and compared to 3 biological replicates of wild type BHT-101 cells.
2020-05-09 | GSE147076 | GEO
Project description:The goal of this study is to assess the role of ASH1 like histone lysine methyltransferase (ASH1L) in the biology of anaplastic thyroid cancer. CRISPR-Cas9 was used to create cell lines derived from BHT-101 anaplastic thyroid cancer cells with premature stop codons prior to the catalytic domain within both alleles of ASH1L. ChIP-seq for H3K36me2, the histone mark catalyzed by ASH1L, was performed on two KO cell lines, and compared to wild type BHT-101 cells.
2020-05-09 | GSE147074 | GEO
Project description:Heat shock proteins (Hsps), in particular Hsp70, play a central role in proteostasis in eukaryotic cells. Due to its chaperone properties, Hsp70 is involved in various processes both after stress and under normal physiological conditions. In contrast to mammals and many Diptera species, members of the Hsp70 family in Drosophila are constitutively synthesized at a low level and undergo dramatic induction after temperature elevation or other forms of stress. In the courtship suppression paradigm used in this study, Drosophila males that have been repeatedly rejected by mated females during courtship are less likely than naive males to court other females. Although numerous genes with known function were identified to play important roles in long-term memory, there is, to the best of our knowledge, no direct evidence implicating Hsp70 in this process. To eliminate this gap, we used D. melanogaster strains containing different hsp70 copy numbers, including strains carrying a deletion of all six hsp70 genes. Our investigations exploring the memory of courtship rejection paradigm demonstrated that a low constitutive level of Hsp70 is required for learning and the formation of short and long-term memories in males. The performed transcriptomic studies demonstrate that males with different hsp70 copy numbers differ significantly in the expression of a few definite groups of genes involved in mating and reproduction, as well as methionine metabolism and immune genes in response to rejection. Specifically, our analysis reveals several major pathways that depend on the presence of hsp70 copies in the genome and apparently participate in memory formation and consolidation, including the cAMP signalling cascade.
2020-06-18 | GSE152647 | GEO
Project description:To determine the phenotypic overlap between restoring miR-101 expression in bladder cancer cells and knocking down EZH2. Either control precursors, pre-miR-101, control siRNA, or EZH2 siRNA were transfected in triplicate into UM-UC-3 cells for 72 hours at a final concentration of 50 nM.
2010-06-25 | E-GEOD-13674 | biostudies-arrayexpress
Project description:From preliminary experiments, HSP70 deficient MEF cells display moderate thermotolerance to a severe heatshock of 45.5 degrees after a mild preshock at 43 degrees, even in the absence of hsp70 protein. We would like to determine which genes in these cells are being activated to account for this thermotolerance. Experiment Overall Design: Two cell lines are analyzed - hsp70 knockout and hsp70 rescue cells. 6 microarrays from the (-/-)knockout cells are analyzed (3 Pretreated vs 3 unheated controls). For the (+/+) rescue cells, 4 microarrays are used (2 pretreated and 2 unheated controls). Cells were plated at 3k/well in a 96 well plate, covered with a gas permeable sealer and heat shocked at 43degrees for 30 minutes at the 20 hr time point. The RNA was harvested at 3hrs after heat treatment.
2009-04-17 | E-GEOD-11120 | biostudies-arrayexpress