title: A RIPK2 inhibitor delays NOD signalling events yet prevents inflammatory cytokine production.
abstract: Intracellular nucleotide binding and oligomerization domain (NOD) receptors recognize antigens including bacterial peptidoglycans and initiate immune responses by triggering the production of pro-inflammatory cytokines through activating NF-κB and MAP kinases. Receptor interacting protein kinase 2 (RIPK2) is critical for NOD-mediated NF-κB activation and cytokine production. Here we develop and characterize a selective RIPK2 kinase inhibitor, WEHI-345, which delays RIPK2 ubiquitylation and NF-κB activation downstream of NOD engagement. Despite only delaying NF-κB activation on NOD stimulation, WEHI-345 prevents cytokine production in vitro and in vivo and ameliorates experimental autoimmune encephalomyelitis in mice. Our study highlights the importance of the kinase activity of RIPK2 for proper immune responses and demonstrates the therapeutic potential of inhibiting RIPK2 in NOD-driven inflammatory diseases.
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Ueli
authors.initials: U
authors.lastname: Nachbur
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Che A
authors.initials: CA
authors.lastname: Stafford
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Bankovacki
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Yifan
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Lisa M
authors.initials: LM
authors.lastname: Lindqvist
authors.affiliation: 1] Department of Disease Biology, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark [2] Ludwig Institute for Cancer Research, Nuffield Department of Clinical Medicine, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Berthe K
authors.initials: BK
authors.lastname: Fiil
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Yelena
authors.initials: Y
authors.lastname: Khakham
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Hyun-Ja
authors.initials: HJ
authors.lastname: Ko
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Jarrod J
authors.initials: JJ
authors.lastname: Sandow
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia [3] Cancer Therapeutics CRC, Bundoora, Victoria 3083, Australia.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Falk
authors.affiliation: ACRF Rational Drug Discovery Centre, St Vincent's Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Australia.
authors.forename: Jessica K
authors.identifier: 0000000287352871
authors.initials: JK
authors.lastname: Holien
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Diep
authors.initials: D
authors.lastname: Chau
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Joanne
authors.initials: J
authors.lastname: Hildebrand
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: James E
authors.initials: JE
authors.lastname: Vince
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Phillip P
authors.initials: PP
authors.lastname: Sharp
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Andrew I
authors.initials: AI
authors.lastname: Webb
authors.affiliation: The Florey Institute of Neuroscience and Mental Health, Melbourne Brain Centre, Austin Campus, 245 Burgundy Street, Heidelberg, Victoria 3084, Australia.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Jackman
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia.
authors.forename: Sabrina
authors.initials: S
authors.lastname: Mühlen
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia.
authors.forename: Catherine L
authors.initials: CL
authors.lastname: Kennedy
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Kym N
authors.initials: KN
authors.lastname: Lowes
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: James M
authors.initials: JM
authors.lastname: Murphy
authors.affiliation: 1] Department of Disease Biology, Novo Nordisk Foundation Center for Protein Research, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark [2] Ludwig Institute for Cancer Research, Nuffield Department of Clinical Medicine, University of Oxford, Oxford OX3 7DQ, UK.
authors.forename: Mads
authors.initials: M
authors.lastname: Gyrd-Hansen
authors.affiliation: 1] ACRF Rational Drug Discovery Centre, St Vincent's Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Australia [2] Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Michael W
authors.identifier: 0000000231011138
authors.initials: MW
authors.lastname: Parker
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia.
authors.forename: Elizabeth L
authors.initials: EL
authors.lastname: Hartland
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Lew
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: David C S
authors.initials: DC
authors.lastname: Huang
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Lessene
authors.affiliation: 1] The Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, Parkville, Victoria 3052, Australia [2] Department of Medical Biology, University of Melbourne, Melbourne, Victoria 3010, Australia.
authors.forename: John
authors.identifier: 0000000276115774
authors.initials: J
authors.lastname: Silke
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D002853:Chromatography, Liquid; D016207:Cytokines / Q000378:metabolism*; D004681:Encephalomyelitis, Autoimmune, Experimental / Q000235:genetics; D005260:Female; D006801:Humans; D007107:Immune System; D007249:Inflammation / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D007371:Interferon-gamma / Q000378:metabolism; D020935:MAP Kinase Signaling System; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D016328:NF-kappa B / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D025801:Ubiquitin / Q000378:metabolism
chemical_list: D016207:Cytokines; D016328:NF-kappa B; D011994:Recombinant Proteins; D025801:Ubiquitin; D007371:Interferon-gamma; D000255:Adenosine Triphosphate; C506253:RIPK2 protein, human; D053475:Receptor-Interacting Protein Serine-Threonine Kinase 2; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C506255:Ripk2 protein, mouse