Active MLKL triggers the NLRP3 inflammasome in a cell-intrinsic manner

Conos, SA; Chen, KW; De Nardo, D; Hara, H; Whitehead, L; Nunez, G; Masters, SL; Murphy, JM; Schroder, K; Vaux, DL; Lawlor, KE; Lindqvist, LM; Vince, JE

Author(s): Conos, SA; Chen, KW; De Nardo, D; Hara, H; Whitehead, L; Nunez, G; Masters, SL; Murphy, JM; Schroder, K; Vaux, DL; Lawlor, KE; Lindqvist, LM; Vince, JE;
Details: Publication Year 2017-02-07,Volume 114,Issue #6,Page E961-E969
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Publication Type: Journal Article
Abstract: Necroptosis is a physiological cell suicide mechanism initiated by receptor-interacting protein kinase-3 (RIPK3) phosphorylation of mixed-lineage kinase domain-like protein (MLKL), which results in disruption of the plasma membrane. Necroptotic cell lysis, and resultant release of proinflammatory mediators, is thought to cause inflammation in necroptotic disease models. However, we previously showed that MLKL signaling can also promote inflammation by activating the nucleotide-binding oligomerization domain (NOD)-like receptor protein 3 (NLRP3) inflammasome to recruit the adaptor protein apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC) and trigger caspase-1 processing of the proinflammatory cytokine IL-1beta. Here, we provide evidence that MLKL-induced activation of NLRP3 requires (i) the death effector four-helical bundle of MLKL, (ii) oligomerization and association of MLKL with cellular membranes, and (iii) a reduction in intracellular potassium concentration. Although genetic or pharmacological targeting of NLRP3 or caspase-1 prevented MLKL-induced IL-1beta secretion, they did not prevent necroptotic cell death. Gasdermin D (GSDMD), the pore-forming caspase-1 substrate required for efficient NLRP3-triggered pyroptosis and IL-1beta release, was not essential for MLKL-dependent death or IL-1beta secretion. Imaging of MLKL-dependent ASC speck formation demonstrated that necroptotic stimuli activate NLRP3 cell-intrinsically, indicating that MLKL-induced NLRP3 inflammasome formation and IL-1beta cleavage occur before cell lysis. Furthermore, we show that necroptotic activation of NLRP3, but not necroptotic cell death alone, is necessary for the activation of NF-kappaB in healthy bystander cells. Collectively, these results demonstrate the potential importance of NLRP3 inflammasome activity as a driving force for inflammation in MLKL-dependent diseases.
Publisher: National Academy of Sciences
Research Division(s): Cell Signalling And Cell Death; Inflammation; Systems Biology And Personalised Medicine
PubMed ID: 28096356
Link To PubMed Central Version: https://www-ncbi-nlm-nih-gov/pmc/articles/PMC5307433/
Publisher's Version: https://doi.org/10.1073/pnas.1613305114
NHMRC Grants: NHMRC/1052598, NHMRC/1051210, NHMRC/1101405, NHMRC/1057905, NHMRC/1020136, NHMRC/1105754, NHMRC/461221,
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2017-04-06 09:27:59

Last Modified: 2018-01-10 09:59:27