Development of an IL-4/IL-13:IL-4Rα in vitro to in vivo Translational Platform in a Model of Respiratory Disease.

Type 2 inflammation drives many chronic respiratory diseases, include asthma and COPD, with IL-4 and IL-13 signalling through IL-4Rα and JAK/STAT6 as central pathway nodes. While the success of dupilumab clinically validates IL-4Rα as a target, there remains a need for efficient, translational assays to discover and characterize next-generation modulators of this pathway. In collaboration with Numerion, Sygnature Discovery has established an integrated in vitro to in vivo platforms that links IL-4/IL-13:IL-4Rα target engagement and signalling with downstream chemokine readouts in a respiratory disease-relevant mouse model.

Using surface plasmon resonance (SPR), we demonstrated high-affinity binding of IL-4 and dupilumab to IL-4Rα, and confirmed disruption of the IL-4:IL-4Rα protein-protein interactions in a sandwich ELISA format. Cellular assays in BEAS-2B lung epithelial cells and mouse splenocytes then captured pathway activation (pSTAT6) and downstream chemokine release (Eotaxin-1), enabling us to benchmark the clinically validated IL-4Rα antibody dupilumab and the JAK inhibitor tofacitinib. Finally, an intranasal IL-13 Balb/c model provided in vivo confirmation that Tofacitinib reduces lung Eotaxin-1 and Eotaxin-2 in a dose-dependent manner with corresponding plasma exposure, complementing a medium-throughput translational cascade suitable for profiling novel IL-4Rα pathway therapeutics in respiratory and other Th2-driven indications.