Bookmark and Share
BioAssay: AID 2604

Quantitative high throughput screen for inhibitors of ROR gamma transcriptional activity: Summary

The retinoic acid-related orphan receptor (ROR) gamma is a transcription factor that has a central role in the differentiation of Th17 cells, a subset of T helper cells that secrete the inflammatory cytokines IL-17, IL-17F, and IL-22. Th17 cells have been implicated in graft versus host disease, autoimmune disease and asthma. ROR gamma is induced in naive T helper cells in the presence of TGF-beta combined with IL-6, IL-21, or IL-23, and thereafter directs the expression of the Th17 lineage cytokines. ..more
_
   
AID: 2604
Data Source: NCGC (RORG954)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2010-03-18
Modify Date: 2010-12-10
Target
Related Experiments
Show more
AIDNameTypeComment
2546VP16 counterscreen qHTS for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Primary counterscreen tested in the VP16 cell line at 6 concentrations from 46 uM to 3 nM
2551qHTS for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Primary screen tested at 6 concentrations from 46 uM to 3 nM
2762Concentration response confirmation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Confirmation assay: retest of selected compounds
2763Concentration response confirmation VP16 counterscreen for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Confirmation assay: retest of selected compounds, VP16 cell line counter screen
489036VP16 selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Selectivity assay of ROR gamma transcriptional activity
489037Secondary confirmation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Secondary confirmation assay
489038DHR3 selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Selectivity assay of ROR gamma transcriptional activity
489039ROR alpha selectivity assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Selectivity assay of ROR gamma transcriptional activity
492954Mouse Th17 T cell differentiation assay for inhibitors of ROR gamma transcriptional activityConfirmatorydepositor-specified cross reference: Selectivity assay of ROR gamma transcriptional activity
492962Mouse Th1 T cell differentiation assay for inhibitors of ROR gamma transcriptional activityOtherdepositor-specified cross reference: Selectivity assay of ROR gamma transcriptional activity
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

MLPCN Grant: R03 DA026211-01
Assay Provider: Dan Littman, New York University

Assay Overview:

The retinoic acid-related orphan receptor (ROR) gamma is a transcription factor that has a central role in the differentiation of Th17 cells, a subset of T helper cells that secrete the inflammatory cytokines IL-17, IL-17F, and IL-22. Th17 cells have been implicated in graft versus host disease, autoimmune disease and asthma. ROR gamma is induced in naive T helper cells in the presence of TGF-beta combined with IL-6, IL-21, or IL-23, and thereafter directs the expression of the Th17 lineage cytokines.

To study the mechanism of action of ROR gamma, two insect cell-based assays, ROR gamma (AID 2551) and VP16 (AID 2546), were screened to find inhibitors of ROR gamma mediated transcription. These Drosophila Schneider cell lines were stably transfected with two vectors: (1), a gene expressing a fusion of the DNA binding domain of Gal4 with the transactivation domain of ROR gamma or VP16 under the control of the metallothionine promoter and (2), a Photinus luciferase reporter gene regulated by the Gal4 binding site enhancer, UAS. Copper addition to the medium induces expression of the Gal4 fusion protein that subsequently activates the UAS-luciferase reporter. Small molecule inhibitors of ROR gamma decrease luciferase reporter activity in the ROR gamma cells but not in the VP16 cells. Small molecule inhibitors that inhibit components common to the ROR gamma and VP16 assays, such as the Gal4 DNA binding domain or the UAS luciferase reporter, are active in both assays and thus scored as nonspecific.
Protocol
Assay Protocol Summary:

Please refer to AIDs 2551 and 2546 for protocol descriptions.
Comment
This summary is written for the purposes of summarizing the probe activities from the project. No probes have yet been declared for this MLPCN project. The current results are from initial screening, and have not yet been validated. MLSCN probes are given a score of 100. Molecules in the prior art are given a score of 80. Other, less active molecules in the same chemical series as the probe molecules are given a score of 50. Molecules pending validation are given a score of 10. Inactive analogues from these series are given a score of 0.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML209
PubChem Substance ID (SID) for probe 1: 99455330
PubChem Compound ID (CID) for probe 1: 53385590
Probe type for probe 1: Antagonist
IC50/EC50 (nM) for probe 1: 460
Fold selectivity for probe 1: >200
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK133441/ (ID: 3025976)
Grant number for probe 1: DA026211-01
NCBI Book chapter title for probe 1: Identification of Potent and Selective ROR? Antagonists
Additional Information
Grant Number: R03 DA026211-01

PageFrom: