title: PMA and crystal-induced neutrophil extracellular trap formation involves RIPK1-RIPK3-MLKL signaling.
abstract: Neutrophil extracellular trap (NET) formation contributes to gout, autoimmune vasculitis, thrombosis, and atherosclerosis. The outside-in signaling pathway triggering NET formation is unknown. Here, we show that the receptor-interacting protein kinase (RIPK)-1-stabilizers necrostatin-1 or necrostatin-1s and the mixed lineage kinase domain-like (MLKL)-inhibitor necrosulfonamide prevent monosodium urate (MSU) crystal- or PMA-induced NET formation in human and mouse neutrophils. These compounds do not affect PMA- or urate crystal-induced production of ROS. Moreover, neutrophils of chronic granulomatous disease patients are shown to lack PMA-induced MLKL phosphorylation. Genetic deficiency of RIPK3 in mice prevents MSU crystal-induced NET formation in vitro and in vivo. Thus, neutrophil death and NET formation may involve the signaling pathway defining necroptosis downstream of ROS production. These data imply that RIPK1, RIPK3, and MLKL could represent molecular targets in gout or other crystallopathies.
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Jyaysi
authors.initials: J
authors.lastname: Desai
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Santhosh V
authors.initials: SV
authors.lastname: Kumar
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Shrikant R
authors.initials: SR
authors.lastname: Mulay
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Lukas
authors.initials: L
authors.lastname: Konrad
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Romoli
authors.affiliation: Department of Internal Medicine 3, University of Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Schauer
authors.affiliation: Department of Internal Medicine 3, University of Erlangen-Nuremberg, Erlangen, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Herrmann
authors.affiliation: Department of Histology, Cytology, Embryology, Danylo Halytsky Lviv National Medical University, Lviv, Ukraine.
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Bilyy
authors.affiliation: Pathologisches Institut, Ludwig-Maximilians Universität, Munich, Germany.
authors.forename: Susanna
authors.initials: S
authors.lastname: Müller
authors.affiliation: Department of Anatomy and Cell Biology, Ludwig-Maximilians Universität, Munich, Germany.
authors.forename: Bastian
authors.initials: B
authors.lastname: Popper
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Daigo
authors.initials: D
authors.lastname: Nakazawa
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Marc
authors.initials: M
authors.lastname: Weidenbusch
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Dana
authors.initials: D
authors.lastname: Thomasova
authors.affiliation: Division of Nephrology and Hypertension, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Krautwald
authors.affiliation: Division of Nephrology and Hypertension, Christian-Albrechts-University, Kiel, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Linkermann
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität München, Munich, Germany.
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Anders
mesh_terms: D000818:Animals; D015153:Blotting, Western; D065206:Extracellular Traps / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000633:toxicity; D011109:Polymethacrylic Acids / Q000633:toxicity; D011494:Protein Kinases / Q000276:immunology / Q000378:metabolism*; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000276:immunology / Q000378:metabolism*; D015398:Signal Transduction / Q000276:immunology*; D014527:Uric Acid / Q000633:toxicity
chemical_list: D011109:Polymethacrylic Acids; C500651:poly(MPC-co-MA); D010767:Phosphorylcholine; D014527:Uric Acid; C570693:MLKL protein, human; C570694:MLKL protein, mouse; D011494:Protein Kinases; C506249:RIPK1 protein, human; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C506251:Ripk1 protein, mouse; C485615:Ripk3 protein, mouse
keywords: Necroptosis; Necrosis; Neutrophil; Neutrophil extracellular trap formation; Receptor-interacting protein kinase