title: Cytotoxicity of crystals involves RIPK3-MLKL-mediated necroptosis.
abstract: Crystals cause injury in numerous disorders, and induce inflammation via the NLRP3 inflammasome, however, it remains unclear how crystals induce cell death. Here we report that crystals of calcium oxalate, monosodium urate, calcium pyrophosphate dihydrate and cystine trigger caspase-independent cell death in five different cell types, which is blocked by necrostatin-1. RNA interference for receptor-interacting protein kinase 3 (RIPK3) or mixed lineage kinase domain like (MLKL), two core proteins of the necroptosis pathway, blocks crystal cytotoxicity. Consistent with this, deficiency of RIPK3 or MLKL prevents oxalate crystal-induced acute kidney injury. The related tissue inflammation drives TNF-α-related necroptosis. Also in human oxalate crystal-related acute kidney injury, dying tubular cells stain positive for phosphorylated MLKL. Furthermore, necrostatin-1 and necrosulfonamide, an inhibitor for human MLKL suppress crystal-induced cell death in human renal progenitor cells. Together, TNF-α/TNFR1, RIPK1, RIPK3 and MLKL are molecular targets to limit crystal-induced cytotoxicity, tissue injury and organ failure.
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Shrikant R
authors.initials: SR
authors.lastname: Mulay
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Jyaysi
authors.initials: J
authors.lastname: Desai
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Santhosh V
authors.initials: SV
authors.lastname: Kumar
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Jonathan N
authors.initials: JN
authors.lastname: Eberhard
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Dana
authors.initials: D
authors.lastname: Thomasova
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Simone
authors.initials: S
authors.lastname: Romoli
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Melissa
authors.initials: M
authors.lastname: Grigorescu
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Onkar P
authors.initials: OP
authors.lastname: Kulkarni
authors.affiliation: Department of Anatomy and Cell Biology, Ludwig-Maximilians Universität, Munich 80336, Germany.
authors.forename: Bastian
authors.initials: B
authors.lastname: Popper
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Vielhauer
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University of Munich, Munich 81377, Germany.|Walter Brendel Center for Experimental Medicine, University of Munich, Munich 81377, Germany.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Zuchtriegel
authors.affiliation: Department of Otorhinolaryngology, Head and Neck Surgery, University of Munich, Munich 81377, Germany.|Walter Brendel Center for Experimental Medicine, University of Munich, Munich 81377, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Reichel
authors.affiliation: Department of Nephropathology, Institute for Pathology, Hannover Medical School, Hannover 30625, Germany.
authors.forename: Jan Hinrich
authors.initials: JH
authors.lastname: Bräsen
authors.affiliation: Excellence Centre for Research, Transfer and High Education for the Development of De Novo Therapies (DENOTHE), University of Florence, Florence 50139, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Romagnani
authors.affiliation: Danylo Halytsky Lviv National Medical University, Lviv 79010, Ukraine.
authors.forename: Rostyslav
authors.initials: R
authors.lastname: Bilyy
authors.affiliation: Department for Internal Medicine, University Hospital Erlangen, Institute for Clinical Immunology, Erlangen 91054, Germany.
authors.forename: Luis E
authors.initials: LE
authors.lastname: Munoz
authors.affiliation: Department for Internal Medicine, University Hospital Erlangen, Institute for Clinical Immunology, Erlangen 91054, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Herrmann
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, Saint Louis, Missouri 63110, USA.|Nephropath, Little Rock, Arkansas 72211, USA.
authors.forename: Helen
authors.initials: H
authors.lastname: Liapis
authors.affiliation: Division of Nephrology and Hypertension, Christian-Albrechts-University, Kiel 24105, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Krautwald
authors.affiliation: Division of Nephrology and Hypertension, Christian-Albrechts-University, Kiel 24105, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Linkermann
authors.affiliation: Medizinische Klinik und Poliklinik IV, Klinikum der Universität, München, Munich 80336, Germany.
authors.forename: Hans-Joachim
authors.initials: HJ
authors.lastname: Anders
mesh_terms: D000818:Animals; D017209:Apoptosis; D002129:Calcium Oxalate / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D002131:Calcium Pyrophosphate / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D006801:Humans; D007674:Kidney Diseases / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009336:Necrosis; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism; D014527:Uric Acid / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*
chemical_list: D014409:Tumor Necrosis Factor-alpha; D002129:Calcium Oxalate; D014527:Uric Acid; C570694:MLKL protein, mouse; D011494:Protein Kinases; D053422:Receptor-Interacting Protein Serine-Threonine Kinases; C485615:Ripk3 protein, mouse; D002131:Calcium Pyrophosphate
references.citation: Science. 2008 May 2;320(5876):674-7
references.pmid: 18403674
references.citation: Cell Microbiol. 2006 Nov;8(11):1812-25
references.pmid: 16824040
references.citation: Immunity. 2008 Nov 14;29(5):807-18
references.pmid: 18993083
references.citation: Science. 2009 Jul 17;325(5938):332-6
references.pmid: 19498109
references.citation: Nature. 2010 Apr 29;464(7293):1357-61
references.pmid: 20428172
references.citation: Urol Res. 2010 Dec;38(6):429-38
references.pmid: 20658131
references.citation: Immunity. 2011 Dec 23;35(6):908-18
references.pmid: 22195746
references.citation: Proc Natl Acad Sci U S A. 2012 Apr 3;109(14):5322-7
references.pmid: 22421439
references.citation: Cell. 2012 May 25;149(5):1060-72
references.pmid: 22632970
references.citation: Arch Pathol Lab Med. 2012 Jul;136(7):713-20
references.pmid: 22742545
references.citation: Stem Cells. 2012 Aug;30(8):1714-25
references.pmid: 22628275
references.citation: J Clin Invest. 2013 Jan;123(1):236-46
references.pmid: 23221343
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 29;110(5):1851-6
references.pmid: 23307811
references.citation: Annu Rev Immunol. 2013;31:73-106
references.pmid: 23215645
references.citation: Autoimmunity. 2013 Aug;46(5):336-41
references.pmid: 23231469
references.citation: Proc Natl Acad Sci U S A. 2013 Jul 16;110(29):12024-9
references.pmid: 23818611
references.citation: Cell Res. 2013 Aug;23(8):994-1006
references.pmid: 23835476
references.citation: Kidney Int. 2013 Nov;84(5):895-901
references.pmid: 23739234
references.citation: Cell Res. 2014 Jan;24(1):105-21
references.pmid: 24366341
references.citation: Cell. 2014 Jan 16;156(1-2):317-31
references.pmid: 24439385
references.citation: Nat Rev Mol Cell Biol. 2014 Feb;15(2):135-47
references.pmid: 24452471
references.citation: N Engl J Med. 2014 Jan 30;370(5):455-65
references.pmid: 24476434
references.citation: Annu Rev Physiol. 2014;76:129-50
references.pmid: 24079414
references.citation: Science. 2014 Mar 21;343(6177):1357-60
references.pmid: 24557836
references.citation: Immunity. 2014 Mar 20;40(3):389-99
references.pmid: 24631154
references.citation: Mol Cell. 2014 Apr 10;54(1):133-46
references.pmid: 24703947
references.citation: Nat Med. 2014 May;20(5):511-7
references.pmid: 24784231
references.citation: J Immunol. 2014 Jun 15;192(12):5671-8
references.pmid: 24799565
references.citation: Nat Rev Immunol. 2014 Nov;14(11):759-67
references.pmid: 25324125
references.citation: Nat Commun. 2015;6:6282
references.pmid: 25693118
references.citation: J Immunol. 2015 Aug 15;195(4):1685-97
references.pmid: 26195813
references.citation: J Vasc Res. 2003 Sep-Oct;40(5):435-41
references.pmid: 14530600
references.citation: Science. 2004 Mar 5;303(5663):1532-5
references.pmid: 15001782
references.citation: Microvasc Res. 1973 May;5(3):384-94
references.pmid: 4709735
references.citation: Methods Cell Biol. 1981;22:97-122
references.pmid: 6267417
references.citation: J Cell Biol. 1988 Oct;107(4):1359-68
references.pmid: 3170633
references.citation: Rheumatol Int. 1997;16(6):241-7
references.pmid: 9106935
references.citation: Int J Urol. 2005 Mar;12(3):290-8
references.pmid: 15828958
references.citation: Am J Pathol. 2005 Jul;167(1):285-98
references.pmid: 15972972
references.citation: Kidney Int. 2005 Oct;68(4):1543-53
references.pmid: 16164631
references.citation: J Leukoc Biol. 2006 Jan;79(1):114-22
references.pmid: 16275892
references.citation: Nat Chem Biol. 2005 Jul;1(2):112-9
references.pmid: 16408008
references.citation: Nature. 2006 Mar 9;440(7081):237-41
references.pmid: 16407889
references.citation: Nat Immunol. 2008 Aug;9(8):847-56
references.pmid: 18604214