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Research at the interface between chemistry and the -omic sciences and systems biology.
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Mol. BioSyst., 2009, 5, 1039 - 1050, DOI: 10.1039/b902021g
A quantitative high-throughput screen for modulators of IL-6 signaling: a model for interrogating biological networks using chemical libraries
Ronald L. Johnson, Ruili Huang, Ajit Jadhav, Noel Southall, Jennifer Wichterman, Ryan MacArthur, Menghang Xia, Kun Bi, John Printen, Christopher P. Austin and James Inglese
Small molecule modulators are critical for dissecting and understanding signaling pathways at the molecular level. Interleukin 6 (IL-6) is a cytokine that signals via the JAK–STAT pathway and is implicated in cancer and inflammation. To identify modulators of this pathway, we screened a chemical collection against an IL-6 responsive cell line stably expressing a
-lactamase reporter gene fused to a sis-inducible element (SIE-bla cells). This assay was optimized for a 1536-well microplate format and screened against 11
693 small molecules using quantitative high-throughput screening (qHTS), a method that assays a chemical library at multiple concentrations to generate titration-response profiles for each compound. The qHTS recovered 564 actives with well-fit curves that clustered into 32 distinct chemical series of 13 activators and 19 inhibitors. A retrospective analysis of the qHTS data indicated that single concentration data at 1.5 and 7.7 
M scored 35 and 71% of qHTS actives, respectively, as inactive and were therefore false negatives. Following counter screens to identify fluorescent and non-selective series, we found four activator and one inhibitor series that modulated SIE-bla cells but did not show similar activity in reporter gene assays induced by EGF and hypoxia. Small molecules within these series will make useful tool compounds to investigate IL-6 signaling mediated by JAK–STAT activation.
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