The effect of fostamatinib, ruxolitinib, and BAY-61-3606 on antiviral innate immune signaling in SUIT-2 cells in response to IFN-a

To dissect the differential impact of fostamatinib and ruxolitinib on interferon (IFN) signaling and related biological processes, we performed global transcriptomic analysis using RNA-sequencing (RNA-seq) on SUIT-2 cells treated with either ruxolitinib, R406, BAY-61-3606, or equal amounts of DMSO (as control) in the presence or absence of type I IFN (IFN-a) stimulation. An additional SYK inhibitor, BAY-61-3606, which does not stimulate VSV replication, was included to distinguish effects due to SYK inhibition from those specific to fostamatinib. Cells were pre-treated with 5 uM of each drug, or DMSO vehicle control, for 16 h, followed by 4h of stimulation with 5,000 U/mL human IFN-a (with drug; 20 h total drug treatment) before total RNA was isolated and analyzed by poly-A enriched RNA sequencing (RNA-seq). This analysis provided a comprehensive overview of how each drug influences gene expression in cells with or without IFN stimulation.