Multivalency enhances the specificity of Fc-cytokine fusions

Abstract

AbstractThe common γ-chain receptor cytokines are promising immune therapies due to their central role in coordinating the proliferation and activity of various immune cell populations. One of these cytokines, interleukin (IL)-2, has potential as a therapy in autoimmunity but is limited in effectiveness by its modest specificity toward regulatory T cells (Tregs). Therapeutic ligands are often made dimeric as antibody Fc fusions to confer desirable pharmacokinetic benefits, with under-explored consequences on signaling. Here, we systematically profiled the signaling responses to a panel of wild type and mutein IL-2 molecules in various Fc fusion configurations. We used a tensor-structured dimensionality reduction scheme to decompose the responses of each cell population to each ligand over a range of time points and cytokine concentrations. We found that dimeric muteins are uniquely specific for Tregs at intermediate ligand concentrations. We then compared signaling response across all treatments to a simple, two-step multivalent binding model. Our model was able to predict cellular responses with high accuracy. Bivalent Fc fusions display enhanced specificity and potency for Tregs through avidity effects toward IL-2Rα. We then utilize our model to identify the potential benefits conferred by valency engineering as an additional mechanism for cytokines with optimized therapeutic benefits. In total, these findings represent a comprehensive analysis of how ligand properties, and their consequent effects on surface receptor-ligand interactions, translate to selective activation of immune cell populations. It also identifies a new route toward engineering even more selective therapeutic cytokines.Significance StatementSignaling in off-target immune cells has hindered the effectiveness of IL-2 as an immunotherapeutic. We show that bivalent IL-2 muteins exhibit more regulatory T cell-selective signaling than monovalent forms. This altered selectivity is explained by altered surface receptor-ligand binding kinetics and can be quantitatively predicted using a multivalent binding model. Finally, our model shows that even more selective IL-2 therapies may be developed by designing cytokines in higher valency formats, revealing valency as an unexplored mechanism for engineering specific IL-2 responses.