qHTS assay to identify small molecule agonists of the peroxisome proliferator-activated receptor delta (PPARd) signaling pathway: Summary
The peroxisome proliferator-activated receptors (PPARs) are lipid-activated transcription factors of the nuclear receptor superfamily with three distinct subtypes namely PPAR alpha, PPAR delta (PPARd) and PPAR gamma. All these subtypes heterodimerizes with Retinoid X receptor (RXR) and have different physiological functions. Although the function of PPAR delta, is less well known until recently, more ..
BioActive Compounds: 97
U.S. Tox21 Program
National Center for Advancing Translational Sciences [NCATS]
NIH Chemical Genomics Center [NCGC]
U.S. Environmental Protection Agency [EPA]
National Institutes of Environmental Health Sciences [NIEHS]
National Toxicology Program [NTP]
U.S. Food and Drug Administration [FDA]
Tox21 Assay Overview:
The peroxisome proliferator-activated receptors (PPARs) are lipid-activated transcription factors of the nuclear receptor superfamily with three distinct subtypes namely PPAR alpha, PPAR delta (PPARd) and PPAR gamma. All these subtypes heterodimerizes with Retinoid X receptor (RXR) and have different physiological functions. Although the function of PPAR delta, is less well known until recently, but the identification of the subtype specific synthetic ligands and the creation of animal models revealed its role in the regulation of cholesterol and lipid metabolism. To identify the compounds that activate PPAR-delta signaling, GeneBLAzer PPAR delta UAS-bla HEK293H cell line (Invitrogen, Carlsbad, CA, USA) containing a beta-lactamase reporter gene under the control of an Upstream Activator Sequence (UAS) was used to screen the Tox21 compound library. The cytotoxicity of the Tox21 compound library against the PPARd-bla cell line was tested in parallel by measuring the cell viability using CellTiter-Glo assay (Promega, Madison, WI) in the same wells. The compounds were also tested for auto fluorescence that may interfere with the biological target readout resulting in potential false positives and/or negatives.
Please refer to other AIDs 743212, 743211, 720687, 720685, 720678 and 720681, for detailed assay protocols.
This summary is written for the purposes of summarizing the compound activities from the project combining the results from both the PPARd agonist mode assay (AID 743212), cell viability counter screen (AID 743211), and auto fluorescence counter screens (AIDs 720687, 720685, 720678 and 720681). For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Potency and efficacy were used for determining relative score. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 5 and 30 determined by phenotype.
Although all reasonable efforts have been made to ensure the accuracy and reliability of the data, caution should be exercised when interpreting the results as artifacts are possible from nonspecific effects such as assay signal interference. The curve fitting and activity calls presented here are based on the NCATS analysis methods. Alternative analysis methods and interpretations of the data are available at EPA (http://actor.epa.gov) and NTP (http://tools.niehs.nih.gov/cebs3/ui/).
* Activity Concentration.
Data Table (Concise)