PXD031319-1

PXD031319 is an original dataset announced via ProteomeXchange.

Title	Novel assays monitoring direct GR protein activity exhibit high predictive power for ligand activity on endogenous GR gene targets
Description	Exogenous glucocorticoids are widely used in the clinic for the treatment of inflammatory disorders and auto-immune diseases. Unfortunately, their use is hampered by many side effects and therapy resistance. Efforts to find more selective glucocorticoid receptor (GR) agonists and modulators (called SEGRAMs), able to separate anti-inflammatory effects via gene suppression from metabolic effects via gene activation, have been unsuccessful so far. In this study, we characterized a set of functionally diverse GR ligands in A549 cells, first using a panel of luciferase-based reporter gene assays evaluating GR-driven gene activation and gene suppression. We expanded this minimal assay set with novel luciferase-based read-outs monitoring GR protein levels, GR dimerization and GR Serine 211 (Ser211) phosphorylation status and compared their outcomes with compound effects on the mRNA levels of known GR target genes in A549 cells and primary hepatocytes. We found that luciferase reporters evaluating GR-driven gene activation and gene repression were not always reliable predictors for effects on endogenous target genes. Remarkably, our novel assay monitoring GR Ser211 phosphorylation levels proved to be the most reliable predictor for compound effects on almost all tested endogenous GR targets, both driven by gene activation and repression. The integration of this novel assay in existing screening platforms may therefore increase chances to find novel GR ligands with an improved therapeutic benefit.
HostingRepository	PRIDE
AnnounceDate	2022-04-11
AnnouncementXML	Submission_2022-04-10_16:50:27.512.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Delphi Van Haver
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-01-30 10:47:02	ID requested
⏵ 1	2022-04-10 16:50:27	announced

Dataset with its publication pending

submitter keyword: Glucocorticoids, glucocorticoid receptor, SEGRAM, drug discovery, inflammation, assay development

Kris Gevaert
contact affiliation	VIB Center for Medical Biotechnology (CMB), Ghent, Belgium Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
contact email	kris.gevaert@vib-ugent.be
lab head
Delphi Van Haver
contact affiliation	VIB Proteomics Core
contact email	delphi.vanhaver@vib-ugent.be
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD031319
     1. Label: PRIDE project
     2. Name: Novel assays monitoring direct GR protein activity exhibit high predictive power for ligand activity on endogenous GR gene targets