Targeting IRAK3 for Degradation to Enhance IL-12 Pro-inflammatory Cytokine Production
Ann Rowley, Brian S. Brown, Mary R. Stofega, Hana Y. Hoh, Rebecca Mathew, Violeta L. Marin, Rongxian Ding, Ryan A. McClure, Fabiola M. Bittencourt, Jun Chen, Tarikere Gururaja, Taisei Kinoshita, Xueqing Wang, Alexey Rivkin, Kevin R. Woller
Abstract
Interleukin-1 receptor-associated kinase 3 (IRAK3) is a pseudokinase mediator in the human inflammatory pathway, and ablation of its function is associated with enhanced antitumor immunity. Traditionally, pseudokinases have eluded "druggability" and have not been considered tractable targets in the pharmaceutical industry. Herein we disclose a CRISPR/Cas9-mediated knockout of IRAK3 in monocyte-derived dendritic cells that results in an increase in IL-12 production upon lipopolysaccharide (LPS) stimulation. Furthermore, we disclose and characterize Degradomer D-1, which displays selective proteasomal degradation of IRAK3 and reproduces the 1L-12p40 increases observed in the CRISPR/Cas9 knockout.