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Cited by 157Cell death mechanisms and regulationLiver Diseases and ImmunityLiver physiology and pathology

The pseudokinase MLKL mediates programmed hepatocellular necrosis independently of RIPK3 during hepatitis

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Günther, Claudia, He, Gui-Wei, Kremer, Andreas E., Murphy, James M., Petrie, Emma J., Amann, Kerstin, Vandenabeele, Peter, Linkermann, Andreas +7 more·2016·Journal of Clinical Investigation
DOI 10.1172/jci87545PMC PMC5096909
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Abstract

Although necrosis and necroinflammation are central features of many liver diseases, the role of programmed necrosis in the context of inflammation-dependent hepatocellular death remains to be fully determined. Here, we have demonstrated that the pseudokinase mixed lineage kinase domain-like protein (MLKL), which plays a key role in the execution of receptor-interacting protein (RIP) kinase-dependent necroptosis, is upregulated and activated in human autoimmune hepatitis and in a murine model of inflammation-dependent hepatitis. Using genetic and pharmacologic approaches, we determined that hepatocellular necrosis in experimental hepatitis is driven by an MLKL-dependent pathway that occurs independently of RIPK3. Moreover, we have provided evidence that the cytotoxic activity of the proinflammatory cytokine IFN-γ in hepatic inflammation is strongly connected to induction of MLKL expression via activation of the transcription factor STAT1. In summary, our results reveal a pathway for MLKL-dependent programmed necrosis that is executed in the absence of RIPK3 and potentially drives the pathogenesis of severe liver diseases.

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References 61 · Cited by 157

Research works printed but not linked: 48

  1. Vercammen D, et al. Dual signaling of the Fas receptor: initiation of both apoptotic and necrotic cell death pathways. J Exp Med. 1998;188(5):919-930.
  2. Zhang H, Zhou X, McQuade T, Li J, Chan FK, Zhang J. Functional complementation between FADD and RIP1 in embryos and lymphocytes. Nature. 2011;471(7338):373-376.
  3. Sun L, et al. Mixed lineage kinase domain-like protein mediates necrosis signaling downstream of RIP3 kinase. Cell. 2012;148(1-2):213-227.
  4. Günther C, et al. Caspase-8 regulates TNF-α- induced epithelial necroptosis and terminal ileitis. Nature. 2011;477(7364):335-339.
  5. Feoktistova M, et al. cIAPs block Ripoptosome formation, a RIP1/caspase-8 containing intracel- lular cell death complex differentially regulated by cFLIP isoforms. Mol Cell. 2011;43(3):449-463.
  6. Cai Z, et al. Plasma membrane transloca- tion of trimerized MLKL protein is required for TNF-induced necroptosis. Nat Cell Biol. 2014;16(1):55-65.
  7. Hildebrand JM, et al. Activation of the pseudo- kinase MLKL unleashes the four-helix bundle domain to induce membrane localization and necroptotic cell death. Proc Natl Acad Sci U S A. 2014;111(42):15072-15077.
  8. Ma X, et al. The oncogenic microRNA miR-21 promotes regulated necrosis in mice. Nat Com- mun. 2015;6:7151.
  9. Linkermann A, et al. Two independent path- ways of regulated necrosis mediate isch- emia-reperfusion injury. Proc Natl Acad Sci U S A. 2013;110(29):12024-12029.
  10. Luedde T, Kaplowitz N, Schwabe RF. Cell death and cell death responses in liver disease: mech- anisms and clinical relevance. Gastroenterology. 2014;147(4):765-783.e4.
  11. Wang H, et al. Mixed lineage kinase domain-like protein MLKL causes necrotic membrane dis- ruption upon phosphorylation by RIP3. Mol Cell. 2014;54(1):133-146.
  12. Ramachandran A, McGill MR, Xie Y, Ni HM, Ding WX, Jaeschke H. Receptor interacting pro- tein kinase 3 is a critical early mediator of acet- aminophen-induced hepatocyte necrosis in mice. Hepatology. 2013;58(6):2099-2108.
  13. Takemoto K, et al. Necrostatin-1 protects against reactive oxygen species (ROS)-induced hepato- toxicity in acetaminophen-induced acute liver failure. FEBS Open Bio. 2014;4:777-787.
  14. Dara L, et al. Receptor interacting protein kinase 1 mediates murine acetaminophen toxicity independent of the necrosome and not through necroptosis. Hepatology. 2015;62(6):1847-1857.
  15. Deutsch M, et al. Divergent effects of RIP1 or RIP3 blockade in murine models of acute liver injury. Cell Death Dis. 2015;6:e1759.
  16. Hussain MJ, Mustafa A, Gallati H, Mowat AP, Mieli-Vergani G, Vergani D. Cellular expression of tumour necrosis factor-alpha and interferon-gam- ma in the liver biopsies of children with chronic liver disease. J Hepatol. 1994;21(5):816-821.
  17. Küsters S, Gantner F, Künstle G, Tiegs G. Inter- feron gamma plays a critical role in T cell-depen- dent liver injury in mice initiated by concanavalin A. Gastroenterology. 1996;111(2):462-471.
  18. Horras CJ, Lamb CL, Mitchell KA. Regulation of hepatocyte fate by interferon-γ. Cytokine Growth Factor Rev. 2011;22(1):35-43.
  19. Siebler J, et al. A key pathogenic role for the STAT1/T-bet signaling pathway in T-cell- mediated liver inflammation. Hepatology. 2003;38(6):1573-1580.
  20. Sun R, et al. STAT1 contributes to dsRNA inhibi- tion of liver regeneration after partial hepatecto- my in mice. Hepatology. 2006;44(4):955-966.
  21. Tanzer MC, et al. Necroptosis signalling is tuned by phosphorylation of MLKL residues outside the pseudokinase domain activation loop. Biochem J. 2015;471(2):255-265.
  22. Tiegs G, Hentschel J, Wendel A. A T cell-depen- dent experimental liver injury in mice inducible by concanavalin A. J Clin Invest. 1992;90(1):196-203.
  23. Murphy JM, et al. The pseudokinase MLKL medi- ates necroptosis via a molecular switch mecha- nism. Immunity. 2013;39(3):443-453.
  24. Degterev A, et al. Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol. 2008;4(5):313-321.
  25. Dillon CP, et al. RIPK1 blocks early postnatal lethality mediated by caspase-8 and RIPK3. Cell. 2014;157(5):1189-1202.
  26. Rickard JA, et al. RIPK1 regulates RIPK3-MLKL- driven systemic inflammation and emergency hematopoiesis. Cell. 2014;157(5):1175-1188.
  27. Duprez L, et al. RIP kinase-dependent necrosis drives lethal systemic inflammatory response syndrome. Immunity. 2011;35(6):908-918.
  28. Rodriguez DA, et al. Characterization of RIPK3-mediated phosphorylation of the activa- tion loop of MLKL during necroptosis. Cell Death Differ. 2016;23(1):76-88.
  29. Pine R, Canova A, Schindler C. Tyrosine phosphor- ylated p91 binds to a single element in the ISGF2/ IRF-1 promoter to mediate induction by IFN alpha and IFN gamma, and is likely to autoregulate the p91 gene. EMBO J. 1994;13(1):158-167.
  30. Küsters S, et al. In vivo evidence for a functional role of both tumor necrosis factor (TNF) recep- tors and transmembrane TNF in experimental hepatitis. Eur J Immunol. 1997;27(11):2870-2875.
  31. Chen X, et al. Translocation of mixed lineage kinase domain-like protein to plasma mem- brane leads to necrotic cell death. Cell Res. 2014;24(1):105-121.
  32. Dondelinger Y, et al. MLKL compromises plasma membrane integrity by binding to phosphatidylinositol phosphates. Cell Rep. 2014;7(4):971-981.
  33. Kang YJ, et al. Regulation of NKT cell-mediated immune responses to tumours and liver inflam- mation by mitochondrial PGAM5-Drp1 signal- ling. Nat Commun. 2015;6:8371.
  34. Newton K, Manning G. Necroptosis and inflam- mation. Annu Rev Biochem. 2016;85:743-763.
  35. Dondelinger Y, Hulpiau P, Saeys Y, Bertrand MJ, Vandenabeele P. An evolutionary perspective on the necroptotic pathway [published online ahead of print June 28, 2016]. Trends Cell Biol. doi: 10.1016/j.tcb.2016.06.004. · DOI · Paper page
  36. Tanzer MC, et al. Evolutionary divergence of the necroptosis effector MLKL. Cell Death Differ. 2016;23(7):1185-1197.
  37. McGill MR, Sharpe MR, Williams CD, Taha M, Curry SC, Jaeschke H. The mechanism under- lying acetaminophen-induced hepatotoxicity in humans and mice involves mitochondrial damage and nuclear DNA fragmentation. J Clin Invest. 2012;122(4):1574-1583.
  38. Vinken M, et al. Primary hepatocytes and their cultures in liver apoptosis research. Arch Toxicol. 2014;88(2):199-212.
  39. Rodriguez DA, et al. Characterization of RIPK3-mediated phosphorylation of the activa- tion loop of MLKL during necroptosis. Cell Death Differ. 2016;23(1):76-88.
  40. McComb S, et al. Type-I interferon signal- ing through ISGF3 complex is required for sustained Rip3 activation and necroptosis in macrophages. Proc Natl Acad Sci U S A. 2014;111(31):E3206-E3213.
  41. García-Buey L, et al. Latent autoimmune hepati- tis triggered during interferon therapy in patients with chronic hepatitis C. Gastroenterology. 1995;108(6):1770-1777.
  42. Newton K, Sun X, Dixit VM. Kinase RIP3 is dis- pensable for normal NF-kappa Bs, signaling by the B-cell and T-cell receptors, tumor necrosis factor receptor 1, and Toll-like receptors 2 and 4. Mol Cell Biol. 2004;24(4):1464-1469.
  43. Bajt ML, Knight TR, Lemasters JJ, Jaeschke H. Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes: protection by N-acetyl cysteine. Toxicol Sci. 2004;80(2):343-349.
  44. Murphy JM, et al. A robust methodology to subclas- sify pseudokinases based on their nucleotide-bind- ing properties. Biochem J. 2014;457(2):323-334.
  45. Babon JJ, Murphy JM. In vitro JAK kinase activ- ity and inhibition assays. Methods Mol Biol. 2013;967:39-55.
  46. Cook WD, et al. RIPK1-and RIPK3-induced cell death mode is determined by target availability. Cell Death Differ. 2014;21(10):1600-1612.
  47. Cartharius K, et al. MatInspector and beyond: pro- moter analysis based on transcription factor bind- ing sites. Bioinformatics. 2005;21(13):2933-2942.
  48. Marinescu VD, Kohane IS, Riva A. MAPPER: a search engine for the computational identifi- cation of putative transcription factor binding sites in multiple genomes. BMC Bioinformatics. 2005;6:79.

Authors
Günther, C., He, G.-W., Kremer, A. E., Murphy, J. M., Petrie, E. J., Amann, K., Vandenabeele, P., Linkermann, A., Poremba, C., Schleicher, U., Dewitz, C., Krautwald, S., Neurath, M. F., Becker, C., & Wirtz, S.
Affiliations
  • Friedrich-Alexander-Universität Erlangen-Nürnberg (DE)
  • The University of Melbourne (AU)
  • Ghent University (BE)
  • University Hospital Schleswig-Holstein (DE)
  • Universitätsklinikum Erlangen (DE)
Corresponding author
Wirtz, Stefan
ORCID
Venue
Journal of Clinical Investigation
Publisher
American Society for Clinical Investigation
Published
2016-10-17
Volume
126
Issue
11
Pages
4346-4360
ISSN
  • 0021-9738 (ISSN-L)
  • 1558-8238
DOI
10.1172/jci87545
PMC
PMC5096909
OpenAlex
W2529871702
Cited by
157
References
61
Citations by year
2026: 3 · 2025: 12 · 2024: 12 · 2023: 13 · 2022: 18 · 2021: 24 · 2020: 17 · 2019: 23 · 2018: 16 · 2017: 16 · 2016: 3
FWCI
10.04
Type
journal article
Language
en
Open access
Locations
  • Journal of Clinical Investigation (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Ghent University Academic Bibliography (Ghent University) (submittedVersion, other-oa)
Funders
  • Fresenius Medical Care North America
  • Deutsche Forschungsgemeinschaft (KFO 257, SFB1181, SPP-1656, BE3686/2, SFB796, KR4391/1‐1)
  • State Government of Victoria
  • Medical Research Council
  • National Health and Medical Research Council (1105754, 9000220, 1057905)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
  • Health Sciences › Medicine › Hepatology › Liver Diseases and Immunity
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
Keywords
MLKL, necroptosis, liver necrosis, autoimmune hepatitis, hepatitis, interferon-gamma, STAT1, liver inflammation, hepatocyte death
MeSH terms
Animals, Humans, Interferon-gamma, Necrosis, Protein Kinases, Mice, Knockout, Hepatitis, Autoimmune, Hepatocytes, STAT1 Transcription Factor, Mice, Receptor-Interacting Protein Serine-Threonine Kinases

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