title: RIP3 induces apoptosis independent of pronecrotic kinase activity.
abstract: Receptor-interacting protein kinase 3 (RIP3 or RIPK3) has emerged as a central player in necroptosis and a potential target to control inflammatory disease. Here, three selective small-molecule compounds are shown to inhibit RIP3 kinase-dependent necroptosis, although their therapeutic value is undermined by a surprising, concentration-dependent induction of apoptosis. These compounds interact with RIP3 to activate caspase 8 (Casp8) via RHIM-driven recruitment of RIP1 (RIPK1) to assemble a Casp8-FADD-cFLIP complex completely independent of pronecrotic kinase activities and MLKL. RIP3 kinase-dead D161N mutant induces spontaneous apoptosis independent of compound, whereas D161G, D143N, and K51A mutants, like wild-type, only trigger apoptosis when compound is present. Accordingly, RIP3-K51A mutant mice (Rip3(K51A/K51A)) are viable and fertile, in stark contrast to the perinatal lethality of Rip3(D161N/D161N) mice. RIP3 therefore holds both necroptosis and apoptosis in balance through a Ripoptosome-like platform. This work highlights a common mechanism unveiling RHIM-driven apoptosis by therapeutic or genetic perturbation of RIP3.
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Pratyusha
authors.initials: P
authors.lastname: Mandal
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Scott B
authors.initials: SB
authors.lastname: Berger
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Sirika
authors.initials: S
authors.lastname: Pillay
authors.affiliation: Department of Pathology, Immunology and Microbiology Program, University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Kenta
authors.initials: K
authors.lastname: Moriwaki
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Chunzi
authors.initials: C
authors.lastname: Huang
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Hongyan
authors.initials: H
authors.lastname: Guo
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Lich
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Finger
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Viera
authors.initials: V
authors.lastname: Kasparcova
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Bart
authors.initials: B
authors.lastname: Votta
authors.affiliation: Molecular Discovery Research, Platform Technologies and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Ouellette
authors.affiliation: Molecular Discovery Research, Platform Technologies and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Bryan W
authors.initials: BW
authors.lastname: King
authors.affiliation: Molecular Discovery Research, Platform Technologies and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: David
authors.initials: D
authors.lastname: Wisnoski
authors.affiliation: Molecular Discovery Research, Platform Technologies and Science, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Ami S
authors.initials: AS
authors.lastname: Lakdawala
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: DeMartino
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Linda N
authors.initials: LN
authors.lastname: Casillas
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Haile
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Clark A
authors.initials: CA
authors.lastname: Sehon
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Marquis
authors.affiliation: Department of Molecular Biosciences, University of Texas at Austin, Austin, TX 78712, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Upton
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Lisa P
authors.initials: LP
authors.lastname: Daley-Bauer
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Linda
authors.initials: L
authors.lastname: Roback
authors.affiliation: Department of Pathology, Immunology and Microbiology Program, University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Nancy
authors.initials: N
authors.lastname: Ramia
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Cole M
authors.initials: CM
authors.lastname: Dovey
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Jan E
authors.initials: JE
authors.lastname: Carette
authors.affiliation: Department of Pathology, Immunology and Microbiology Program, University of Massachusetts Medical School, Worcester, MA 01605, USA.
authors.forename: Francis Ka-Ming
authors.initials: FK
authors.lastname: Chan
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: John
authors.initials: J
authors.lastname: Bertin
authors.affiliation: Pattern Recognition Receptor Discovery Performance Unit, Immuno-Inflammation Therapeutic Area, GlaxoSmithKline, Collegeville, PA 19426, USA.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Gough
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA. Electronic address: mocarski@emory.edu.
authors.forename: Edward S
authors.initials: ES
authors.lastname: Mocarski
authors.affiliation: Department of Microbiology and Immunology, Emory Vaccine Center, Emory University School of Medicine, Atlanta, GA 30322, USA. Electronic address: wkaiser@emory.edu.
authors.forename: William J
authors.initials: WJ
authors.lastname: Kaiser
mesh_terms: D000818:Animals; D017209:Apoptosis; D053181:Caspase 8 / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism; D055879:Gene Knock-In Techniques; D019073:HT29 Cells; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D041681:NIH 3T3 Cells; D009336:Necrosis / Q000201:enzymology; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000378:metabolism; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*
chemical_list: C119724:AGFG1 protein, human; C505533:FADD protein, human; D053200:Fas-Associated Death Domain Protein; D028861:Nuclear Pore Complex Proteins; D047428:Protein Kinase Inhibitors; D016601:RNA-Binding Proteins; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C505530:CASP8 protein, human; D053181:Caspase 8