title: PIKfyve, a class III PI kinase, is the target of the small molecular IL-12/IL-23 inhibitor apilimod and a player in Toll-like receptor signaling.
abstract: Toll-like receptor (TLR) signaling is a key component of innate immunity. Aberrant TLR activation leads to immune disorders via dysregulation of cytokine production, such as IL-12/IL-23. Herein, we identify and characterize PIKfyve, a lipid kinase, as a critical player in TLR signaling using apilimod as an affinity tool. Apilimod is a potent small molecular inhibitor of IL-12/IL-23 with an unknown target and has been evaluated in clinical trials for patients with Crohn's disease or rheumatoid arthritis. Using a chemical genetic approach, we show that it binds to PIKfyve and blocks its phosphotransferase activity, leading to selective inhibition of IL-12/IL-23p40. Pharmacological or genetic inactivation of PIKfyve is necessary and sufficient for suppression of IL-12/IL-23p40 expression. Thus, we have uncovered a phosphoinositide-mediated regulatory mechanism that controls TLR signaling.
authors.affiliation: Novartis Institutes for Biomedical Research, 250 Massachusetts Avenue, Cambridge, MA 02139, USA.
authors.forename: Xinming
authors.initials: X
authors.lastname: Cai
authors.forename: Yongyao
authors.initials: Y
authors.lastname: Xu
authors.forename: Atwood K
authors.initials: AK
authors.lastname: Cheung
authors.forename: Ronald C
authors.initials: RC
authors.lastname: Tomlinson
authors.forename: Abel
authors.initials: A
authors.lastname: Alcázar-Román
authors.forename: Leon
authors.initials: L
authors.lastname: Murphy
authors.forename: Andreas
authors.initials: A
authors.lastname: Billich
authors.forename: Bailin
authors.initials: B
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Feng
authors.forename: Martin
authors.initials: M
authors.lastname: Klumpp
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Rondeau
authors.forename: Aleem N
authors.initials: AN
authors.lastname: Fazal
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Wilson
authors.forename: Vic
authors.initials: V
authors.lastname: Myer
authors.forename: Gerard
authors.initials: G
authors.lastname: Joberty
authors.forename: Tewis
authors.initials: T
authors.lastname: Bouwmeester
authors.forename: Mark A
authors.initials: MA
authors.lastname: Labow
authors.forename: Peter M
authors.initials: PM
authors.lastname: Finan
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Porter
authors.forename: Hidde L
authors.initials: HL
authors.lastname: Ploegh
authors.forename: Daniel
authors.initials: D
authors.lastname: Baird
authors.forename: Pietro
authors.initials: P
authors.lastname: De Camilli
authors.forename: John A
authors.initials: JA
authors.lastname: Tallarico
authors.forename: Qian
authors.initials: Q
authors.lastname: Huang
mesh_terms: D000818:Animals; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D006835:Hydrazones; D018664:Interleukin-12 / Q000037:antagonists & inhibitors*; D053759:Interleukin-23 / Q000037:antagonists & inhibitors*; D051379:Mice; D009025:Morpholines / Q000378:metabolism / Q000494:pharmacology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011485:Protein Binding; D011743:Pyrimidines; D015398:Signal Transduction / Q000187:drug effects*; D013379:Substrate Specificity; D051193:Toll-Like Receptors / Q000378:metabolism*; D014227:Triazines / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D004791:Enzyme Inhibitors; D006835:Hydrazones; D018664:Interleukin-12; D053759:Interleukin-23; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011743:Pyrimidines; D051193:Toll-Like Receptors; D014227:Triazines; C492139:PIKFYVE protein, human; C504227:apilimod
references.citation: Cell. 1993 May 7;73(3):585-96
references.pmid: 8387896
references.citation: Cell. 2010 Mar 19;140(6):805-20
references.pmid: 20303872
references.citation: Nature. 2007 Jul 5;448(7149):68-72
references.pmid: 17572665
references.citation: Science. 2000 Mar 17;287(5460):1964-9
references.pmid: 10720315
references.citation: J Exp Med. 2008 Feb 18;205(2):315-22
references.pmid: 18227218
references.citation: J Biol Chem. 2001 Jul 13;276(28):26141-7
references.pmid: 11285266
references.citation: Nat Rev Drug Discov. 2010 Apr;9(4):293-307
references.pmid: 20380038
references.citation: EMBO Rep. 2008 Feb;9(2):164-70
references.pmid: 18188180
references.citation: Nature. 1994 Jun 30;369(6483):756-8
references.pmid: 8008069
references.citation: Proc Natl Acad Sci U S A. 2007 Oct 30;104(44):17518-23
references.pmid: 17956977
references.citation: Science. 2003 Aug 1;301(5633):640-3
references.pmid: 12855817
references.citation: IDrugs. 2007 Jan;10(1):53-9
references.pmid: 17187316
references.citation: Inflamm Bowel Dis. 2010 Jul;16(7):1209-18
references.pmid: 19918967
references.citation: PLoS One. 2012;7(4):e35069
references.pmid: 22493730
references.citation: Nature. 2009 Oct 1;461(7264):614-20
references.pmid: 19759537
references.citation: Chembiochem. 2005 Mar;6(3):523-6
references.pmid: 15696508
references.citation: J Biol Chem. 2011 Apr 15;286(15):13404-13
references.pmid: 21349843
references.citation: EMBO J. 2008 Dec 17;27(24):3221-34
references.pmid: 19037259
references.citation: J Cell Sci. 2006 Oct 1;119(Pt 19):3944-57
references.pmid: 16954148
references.citation: Immunol Rev. 2004 Dec;202:96-105
references.pmid: 15546388
references.citation: Nature. 2008 Mar 13;452(7184):234-8
references.pmid: 18305481
references.citation: Proc Natl Acad Sci U S A. 1994 Dec 20;91(26):12574-8
references.pmid: 7809080
references.citation: Nat Immunol. 2002 Sep;3(9):875-81
references.pmid: 12154357
references.citation: Endocrinology. 2002 Dec;143(12):4742-54
references.pmid: 12446602
references.citation: Annu Rev Immunol. 1998;16:495-521
references.pmid: 9597139
references.citation: Biochem Biophys Res Commun. 2009 May 8;382(3):566-70
references.pmid: 19289105
references.citation: J Biol Chem. 2007 Aug 17;282(33):23878-91
references.pmid: 17556371
references.citation: Nat Immunol. 2006 Feb;7(2):156-64
references.pmid: 16415873
references.citation: Cell. 1994 Jul 15;78(1):35-43
references.pmid: 7518356
references.citation: Biochem J. 2009 Apr 1;419(1):1-13
references.pmid: 19272020
references.citation: J Exp Med. 2002 Apr 1;195(7):953-8
references.pmid: 11927638
references.citation: Nat Immunol. 2008 Oct;9(10):1157-64
references.pmid: 18758466
references.citation: Cell. 2006 Jun 2;125(5):943-55
references.pmid: 16751103
references.citation: Mol Cell Biol. 2004 Dec;24(23):10437-47
references.pmid: 15542851
references.citation: Cell. 2010 Mar 19;140(6):935-50
references.pmid: 20303881
references.citation: Annu Rev Med. 2009;60:97-110
references.pmid: 18976050
references.citation: Cell Biol Int. 2008 Jun;32(6):591-604
references.pmid: 18304842
references.citation: Nature. 1997 Jul 24;388(6640):394-7
references.pmid: 9237759
references.citation: Immunol Rev. 2007 Dec;220:251-69
references.pmid: 17979852
references.citation: Chem Soc Rev. 2005 Jun;34(6):472-82
references.pmid: 16137160
references.citation: Blood. 2007 Feb 1;109(3):1156-64
references.pmid: 17053051
references.citation: Hum Mol Genet. 2009 Dec 15;18(24):4868-78
references.pmid: 19793721