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BioAssay: AID 1808

Summary of probe development efforts to identify agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPAR gamma)

Name: Summary of probe development efforts to identify agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPAR gamma) ..more
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AID: 1808
Data Source: The Scripps Research Institute Molecular Screening Center (PPARGSRC1_AG_LEADS_SUMMARY)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2009-06-08
Modify Date: 2011-07-27
Targets
Sequence: peroxisome proliferative activated receptor gamma isoform 2 [Homo sapiens]
Description ..   
Protein Family: The ligand binding domain of peroxisome proliferator-activated receptors

Gene:PPARG     Related Protein 3D Structures     More BioActivity Data..


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Depositor Specified Assays
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AIDNameTypeProbeComment
631Primary biochemical High Throughput Screening assay for agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPARgamma)screening Primary HTS in singlicate to identify SRC1 agonists.
1051Measurement of TR-FRET detection format artefact in the screen for agonists of steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPARgamma)other Counter screen in triplicate to identify possible fluorescent artifacts.
1300Confirmation biochemical High Throughput Screening assay for agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPARgamma)screening Confirmation of hit activity in triplicate.
1319Dose response biochemical High Throughput Screening assay for agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPARgamma)confirmatory Titration assays in triplicate to determine potency of selective compounds.
1679TR-FRET dose response biochemical High Throughput Screening assay for agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPAR gamma): non-selective agonistsconfirmatory Titration assays in triplicate to determine potency of non-selective compounds.
504446TR-FRET-based biochemical dose response competitive binding lanthascreen assay for partial agonists of the peroxisome proliferator-activated receptor gamma (PPARg)confirmatory Dose response Lanthascreen (PPARG partial agonists in triplicate)
504447Luminescence-based cell-based assay provider high throughput dose response assay for partial agonists of the peroxisome proliferator-activated receptor gamma (PPARg)confirmatory CBI Dose response (PPARG partial agonists in triplicate)
504452Luminescence-based cell-based primary high throughput screening assay to identify partial agonists of the peroxisome proliferator-activated receptor gamma (PPARg)screening Primary screen (PPARG partial agonists in duplicate)
504453Fluorescence-polarization-based biochemical polarscreen dose response binding assay for partial agonists of the peroxisome proliferator-activated receptor gamma (PPARg)confirmatory Dose response Polarscreen (PPARG partial agonists in triplicate)
504735Late stage assay provider counterscreen for partial agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Luminescence-based cell-based dose response assay for partial agonists of the peroxisome proliferator-activated receptor alpha (PPARA).confirmatory Late stage dose response counterscreen (PPARA partial agonists in triplicate)
504938Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): chemiluminescence-based biochemical western blot assay for inhibitors of phosphorylation of PPARG protein levelsother1 Late stage counterscreen western blot assay (phosphorylation of PPARG protein levels inhibitors)
504939Late stage assay provider results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Luminescence-based cell-based dose response assay to identify transactivation inhibitor of PPRE (PPARG response element)confirmatory Late stage dose response assay (PPRE transactivation inhibitors)
504943Late stage assay provider results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): TR-FRET-based biochemical dose response competitive binding lanthascreen assay for non-agonists of the PPARgconfirmatory Late stage dose response lanthascreen (PPARg non-agonists)
540282Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assessPPAR-gamma (PPARG) gene expression changes associated with osteoblast differentiationother Late stage counterscreen (PPARG gene expression changes in triplicate)
540283Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess receptor activator of nuclear factor kappa-B ligand (RANKL) gene expression changes associated with osteoblast differentiationother Late stage counterscreen (RANKL gene expression changes in triplicate)
540284Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess type I collagen (col1) gene expression changes associated with osteoblast differentiationother Late stage counterscreen (col1 gene expression changes in triplicate)
540285Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess alkaline phosphatase (Alp) gene expression changes associated with osteoblast differentiationother Late stage counterscreen (Alp gene expression changes in triplicate)
540286Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess AP2 (fatty acid binding protein 4) gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (AP2 gene expression changes in triplicate)
540287Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess C/EBP-alpha gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (C/EBP-alpha gene expression changes in triplicate)
540289Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess PPAR-gamma gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (PPAR-gamma gene expression changes in triplicate)
540290Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess CD36 gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (CD36 gene expression changes in triplicate)
540291Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess Lpl gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (Lpl gene expression changes in triplicate)
540292Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess Fasn gene expression changes associated with adipocyte differentiation in 3T3-L1 cells.other Late stage counterscreen (Fasn gene expression changes in triplicate)
540293Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): Leptin-deficient ob/ob mouse model studies to assess the effect of probe candidate on fasting blood glucoseother1 Late stage counterscreen (fasting blood glucose)
540294Late stage assay provider counterscreen results from the probe development effort to identify non-agonists of the peroxisome proliferator-activated receptor gamma (PPARg): fluorescence-based cell-based quantitative polymerase chain reaction assay to assess Glut4 gene expression changes associated with adipocyte differentiation in 3T3-L1 cellsother Late stage counterscreen (Glut4 gene expression changes in triplicate)
Description:
Source (MLPCN Center Name): The Scripps Research Institute Molecular Screening Center (SRIMSC)
Center Affiliation: The Scripps Research Institute (TSRI)
Assay Provider: Pat Griffin, TSRI
Network: Molecular Library Probe Production Center Network (MLPCN)
Grant Proposal Number: 1 X01 MH079861-01
Grant Proposal PI: Patrick Griffin, TSRI
External Assay ID: PPARGSRC1_AG_LEADS_SUMMARY

Name: Summary of probe development efforts to identify agonists of the steroid receptor coactivator 1 (SRC-1) recruitment by the peroxisome proliferator-activated receptor gamma (PPAR gamma)

Description:

Peroxisome proliferator-activated receptors (PPARs) belong to the nuclear receptor superfamily and are lipid sensors functioning as ligand-dependent transcription factors regulating gene expression patterns of diverse biological processes (1, 2). PPARs play a critical role in metabolic processes such as glucose metabolism, lipid metabolism, and have been implicated in anti-atherogenic, anti-inflammatory as well as anti-hypertensive functions (3). Like other nuclear receptors, PPARs act as agonist-activated transcription factors, regulating specific target gene transcription. PPARs have been shown to respond to small molecules and are well-documented for therapeutic actions triggered by synthetic agonists (4-6). Among the three isoforms of PPAR identified, PPAR gamma (NR1C3) is implicated in several important disorders such as atherosclerosis, diabetes, obesity and cancer, providing strong justification for the search for specific PPARg agonists that can be used to treat these pathologies. However, the clinical use of PPARg agonists has been associated with adverse effects that are mainly caused by the concomitant activation of various target genes implicated in different physiological pathways. These side effects include weight gain through increased adipogenesis, renal fluid retention and plasma volume expansion, as well as toxic effects in the liver (7). To design safer and more selective PPARg agonists, the different physiological pathways triggered by PPARg activation have to be decoupled. Therefore, screening for agonists that favor specifically the association of a given cofactor will provide useful chemical tools for probing PPARg/coactivator interactions, helping the design of safer PPARg agonists.

Summary of Probe Development Effort:

Following primary HTS in singlicate to identify SRC1 agonists (AID 631), counterscreening in triplicate to identify possible fluorescent artifacts (AID 1051), confirmation of hit activity in triplicate (AID 1300), followed by titration assays in triplicate to determine potency of selective (AID 1319) and non-selective compounds (AID 1679), certain compounds were identified as possible candidates for probe development. A probe development effort is currently underway at the SRIMSC.

References:
1. Chawla, A., et al., Nuclear receptors and lipid physiology: Opening the X-files. Science, 2001. 294(5548): p. 1866-1870.
2. Krey, G., et al., Fatty acids, eicosanoids, and hypolipidemic agents identified as ligands of peroxisome proliferator-activated receptors by coactivator-dependent receptor ligand assay. Molecular Endocrinology, 1997. 11(6): p. 779-791.
3. Bishop-Bailey, D., T. Hla, and T.D. Warner, Intimal smooth muscle cells as a target for peroxisome proliferator-activated receptor-gamma ligand therapy. Circ Res, 2002. 91(3): p. 210-7.
4. Evans, R.M., G.D. Barish, and Y.X. Wang, PPARs and the complex journey to obesity. Nat Med, 2004. 10(4): p. 355-61. PMID: 15057233.
5. Staels, B., et al., Mechanism of action of fibrates on lipid and lipoprotein metabolism. Circulation, 1998. 98(19): p. 2088-93.
6. Barish, G.D., V.A. Narkar, and R.M. Evans, PPAR delta: a dagger in the heart of the metabolic syndrome. J Clin Invest, 2006. 116(3): p. 590-7.
7. Berger, J.P., T.E. Akiyama, and P.T. Meinke, PPARs: therapeutic targets for metabolic disease. Trends Pharmacol Sci, 2005. 26(5): p. 244-51.

Keywords:

Summary AID, PPAR gamma, PPARg, PPARG1, PPARG2, SRC-1, SRC1, nuclear receptor coactivator 1, NCOA1, NCoA-1, PAX3/NCOA1 fusion protein, F-SRC-1, RIP160, KAT13A, agonist, agonism, activator, selective, non-selective, time-resolved fluorescence energy transfer, TR-FRET, FRET, 1536-well, HTS, High Throughput Screening, dose response, Scripps, Scripps Florida, Scripps Research Institute Molecular Screening Center, SRIMSC, Molecular Libraries Probe Production Centers Network, MLPCN.
Protocol
Please see AIDs 631, 1051, 1300, 1319, and 1679 for all protocols performed in this probe development effort.
Comment
This probe development project is still underway at the SRIMSC.

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