A RIPK2 inhibitor delays NOD signalling events yet prevents inflammatory cytokine production

Ueli Nachbur; Che A Stafford; Aleksandra Bankovacki; Yifan Zhan; Lisa M Lindqvist; Berthe K Fiil; Yelena Khakham; Hyun-Ja Ko; Jarrod J Sandow; Hendrik Falk; Jessica K Holien; Diep Chau; Joanne Hildebrand; James E Vince; Phillip P Sharp; Andrew I Webb; Katherine A Jackman; Sabrina Mühlen; Catherine L Kennedy; Kym N Lowes; James M Murphy; Mads Gyrd-Hansen; Michael W Parker; Elizabeth L Hartland; Andrew M Lew; David C S Huang; Guillaume Lessene; John Silke

doi:10.1038/ncomms7442

A RIPK2 inhibitor delays NOD signalling events yet prevents inflammatory cytokine production

Nat Commun. 2015 Mar 17:6:6442. doi: 10.1038/ncomms7442.

Authors

Ueli Nachbur¹, Che A Stafford¹, Aleksandra Bankovacki¹, Yifan Zhan¹, Lisa M Lindqvist¹, Berthe K Fiil², Yelena Khakham¹, Hyun-Ja Ko¹, Jarrod J Sandow¹, Hendrik Falk³, Jessica K Holien⁴, Diep Chau¹, Joanne Hildebrand¹, James E Vince¹, Phillip P Sharp¹, Andrew I Webb¹, Katherine A Jackman⁵, Sabrina Mühlen⁶, Catherine L Kennedy⁶, Kym N Lowes¹, James M Murphy¹, Mads Gyrd-Hansen², Michael W Parker⁷, Elizabeth L Hartland⁶, Andrew M Lew¹, David C S Huang¹, Guillaume Lessene¹, John Silke¹

Affiliations

¹ 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.
² 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.
³ 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.
⁴ ACRF Rational Drug Discovery Centre, St Vincent's Institute of Medical Research, 9 Princes Street, Fitzroy, Victoria 3065, Australia.
⁵ The Florey Institute of Neuroscience and Mental Health, Melbourne Brain Centre, Austin Campus, 245 Burgundy Street, Heidelberg, Victoria 3084, Australia.
⁶ Department of Microbiology and Immunology, University of Melbourne, Victoria 3010, Australia.
⁷ 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.

PMID: 25778803
DOI: 10.1038/ncomms7442

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine Triphosphate / chemistry
Animals
Chromatography, Liquid
Cytokines / metabolism*
Encephalomyelitis, Autoimmune, Experimental / genetics
Female
Humans
Immune System
Inflammation / metabolism*
Inhibitory Concentration 50
Interferon-gamma / metabolism
MAP Kinase Signaling System
Male
Mice
Mice, Inbred C57BL
Mice, Inbred NOD
NF-kappa B / metabolism
Protein Binding
Protein Conformation
Receptor-Interacting Protein Serine-Threonine Kinase 2 / antagonists & inhibitors
Receptor-Interacting Protein Serine-Threonine Kinase 2 / metabolism
Receptor-Interacting Protein Serine-Threonine Kinases / antagonists & inhibitors
Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
Recombinant Proteins / metabolism
Signal Transduction
Tandem Mass Spectrometry
Ubiquitin / metabolism

Substances

Cytokines
NF-kappa B
Recombinant Proteins
Ubiquitin
Interferon-gamma
Adenosine Triphosphate
RIPK2 protein, human
Receptor-Interacting Protein Serine-Threonine Kinase 2
Receptor-Interacting Protein Serine-Threonine Kinases
Ripk2 protein, mouse