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Updated: Mar 1, 2026
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Detection and Isolation of Viable Mouse IL-17-Secreting T Cells
Published on: December 18, 2008
11.7K
High Throughput Screening Assay to Identify Modulators of IL-17 Expression
Mohamed Boudjelal1, Ana Maria Ruiz-Avendano2, Gonzalo Colmenarejo3
1King Abdullah International Medical Research Centre, Ministry of National Guard Health Affairs, Riyadh, Saudi Arabia.
Combinatorial Chemistry & High Throughput Screening
|June 3, 2017
Summary
We developed a new assay to find small molecule inhibitors of RORγt and IL-17. This parallel screening approach identified promising compounds, including novel IL-17 modulators.
Area of Science:
- Pharmacology and Immunology
- Drug Discovery
- Molecular Biology
Background:
- Retinoid-related Orphan Receptor gamma t (RORγt) is a key regulator of the IL-17 pathway, implicated in autoimmune diseases.
- Identifying small molecule inhibitors of RORγt and IL-17 is crucial for developing new therapeutics.
- Existing screening methods may not efficiently identify compounds with diverse mechanisms of action.
Purpose of the Study:
- To develop and validate a novel RORγt-enhanced IL-17F promoter-luciferase reporter assay for high-throughput screening (HTS).
- To identify cell-permeable small-molecule inhibitors of RORγt and IL-17 using a parallel screening approach.
- To evaluate the mode of action of identified inhibitors, distinguishing direct RORγt modulators from other IL-17 pathway inhibitors.
Main Methods:
- Development of a stable Jurkat T-cell line expressing RORγt for the IL-17F promoter-luciferase assay, miniaturized for HTS in 1536-well plates.
- Parallel screening of a >350k compound library using the luciferase reporter assay and a RORγt time-resolved Förster resonance energy transfer (TR-FRET) binding assay.
- Rigorous triaging of hits, including in-silico filtering, selectivity screening (IL-2 reporter assay), and phenotypic profiling in a PBMC IL-17A production assay.
Main Results:
- Successful identification of cell-permeable RORγt antagonists active in both reporter and binding assays.
- Discovery of a distinct set of compounds inhibiting IL-17 reporter activity without direct RORγt modulation.
- Identification of promising small molecules inhibiting IL-17 via potentially novel pathways after rigorous triaging.
Conclusions:
- The developed RORγt-luciferase reporter assay is effective for HTS of small molecule RORγt/IL-17 inhibitors.
- A parallel screening strategy combining multiple assay formats and in-silico triaging accelerates the identification of modulators with diverse mechanisms.
- This approach enables the discovery of novel IL-17 pathway inhibitors with therapeutic potential.
