In Vivo Inhibition of RIPK2 Kinase Alleviates Inflammatory Disease

• Published in: The Journal of biological chemistry Vol. 289; no. 43; pp. 29651 - 29664
• Main Authors: Tigno-Aranjuez, Justine T., Benderitter, Pascal, Rombouts, Frederik, Deroose, Frederik, Bai, XiaoDong, Mattioli, Benedetta, Cominelli, Fabio, Pizarro, Theresa T., Hoflack, Jan, Abbott, Derek W.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 24-10-2014; American Society for Biochemistry and Molecular Biology
• Subjects: Acetylmuramyl-Alanyl-Isoglutamine; Animals; Crohn Disease - drug therapy; Crohn Disease - enzymology; Crohn Disease - pathology; Disease Models, Animal; Down-Regulation - drug effects; Down-Regulation - genetics; Gene Expression Profiling; Genetic Markers; HEK293 Cells; Humans; Ileitis - drug therapy; Ileitis - enzymology; Ileitis - pathology; Inflammation - drug therapy; Inflammation - enzymology; Inflammation - pathology; Macrophages - drug effects; Macrophages - enzymology; Macrophages - pathology; Mice, Inbred C57BL; Peritonitis - chemically induced; Peritonitis - pathology; Protein Kinase Inhibitors - chemistry; Protein Kinase Inhibitors - pharmacology; Protein Kinase Inhibitors - therapeutic use; Receptor-Interacting Protein Serine-Threonine Kinase 2 - antagonists & inhibitors; Receptor-Interacting Protein Serine-Threonine Kinase 2 - metabolism; Signal Transduction; Substrate Specificity - drug effects; Transcriptome - genetics; Treatment Outcome; Immunology; Signal Transduction; Dual Specificity Kinase; Nod-like Receptor (NLR); Innate Immunity; Enzyme Inhibitor
• DOI: 10.1074/jbc.M114.591388
• EISSN: 1083-351X
• ISSN: 0021-9258

The RIPK2 kinase transduces signaling downstream of the intracellular peptidoglycan sensors NOD1 and NOD2 to promote a productive inflammatory response. However, excessive NOD2 signaling has been associated with numerous diseases, including inflammatory bowel disease (IBD), sarcoidosis and inflammatory arthritis, making pharmacologic inhibition of RIPK2 an appealing strategy. In this work, we report the generation, identification, and evaluation of novel RIPK2 specific inhibitors. These compounds potently inhibit the RIPK2 tyrosine kinase activity in in vitro biochemical assays and cellular assays, as well as effectively reduce RIPK2-mediated effects in an in vivo peritonitis model. In conjunction with the development of these inhibitors, we have also defined a panel of genes whose expression is regulated by RIPK2 kinase activity. Such RIPK2 activation markers may serve as a useful tool for predicting settings likely to benefit from RIPK2 inhibition. Using these markers and the FDA-approved RIPK2 inhibitor Gefitinib, we show that pharmacologic RIPK2 inhibition drastically improves disease in a spontaneous model of Crohn Disease-like ileitis. Furthermore, using novel RIPK2-specific inhibitors, we show that cellular recruitment is inhibited in an in vivo peritonitis model. Altogether, the data presented in this work provides a strong rationale for further development and optimization of RIPK2-targeted pharmaceuticals and diagnostics.