[en] Inflammation within the CNS is a major component of many neurodegenerative diseases. A characteristic feature is the generation of microglia-derived factors that play an essential role in the immune response. IL-1β is a pro-inflammatory cytokine released by activated microglia, able to exacerbate injury at elevated levels. In the presence of caspase-1, pro-IL-1β is cleaved to the mature cytokine following NOD-like receptor pyrin domain containing 3 (NLRP3) inflammasome activation. Growing evidence suggests that ceramide plays a critical role in NLRP3 inflammasome assembly, however, the relationship between ceramide and inflammasome activation in microglia remains unknown. Here, we investigated potential mechanistic links between ceramide as a modulator of NLRP3 inflammasome assembly and the resulting secretion of IL-1β using small bioactive enzyme stimulators and inhibitors of ceramide signaling in wild-type and apoptosis-associated speck-like protein containing a CARD knockout (ASC-/- ) primary microglia. To induce the expression of inflammasome components, microglia were primed prior to experiments. Treatment with sodium palmitate (PA) induced de novo ceramide synthesis via modulation of its synthesizing protein serine palmitoyl transferase resulting in increased IL-1β secretion in microglia. Exposure of microglia to the serine palmitoyl transferase-inhibitor l-cycloserine significantly prevented PA-induced IL-1β secretion. Application of the ceramide analogue C2 and the sphingosine-1-phosphate-receptor agonist Fingolimod (FTY720) up-regulated levels of IL-1β and cleaved caspase-1 in wild-type microglia, whereas ASC-/- microglia were unaffected. HPA-12 inhibition of ceramide transport did not affect inflammasome activation. Taken together, our findings reveal a critical role for ceramide as a positive modulator of NLRP3 inflammasome assembly and the resulting release of IL-1β.
Disciplines :
Neurology
Author, co-author :
Scheiblich, Hannah; Department of Neurodegenerative Disease and Gerontopsychiatry/Neurology, University of Bonn - Medical Center, Bonn, Germany
Schlütter, Anna; Department of Neurodegenerative Disease and Gerontopsychiatry/Neurology, University of Bonn - Medical Center, Bonn, Germany ; Department of Neuroscience, Maastricht University, Maastricht, The Netherlands
Golenbock, Douglas T; Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, USA
Latz, Eicke; Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, USA ; Institute of Innate Immunity, University of Bonn, Bonn, Germany ; German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany
Martinez-Martinez, Pilar; Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, USA
HENEKA, Michael ; Department of Neurodegenerative Disease and Gerontopsychiatry/Neurology, University of Bonn - Medical Center, Bonn, Germany ; Department of Infectious Diseases and Immunology, University of Massachusetts Medical School, Worcester, Massachusetts, USA ; German Center for Neurodegenerative Diseases (DZNE), Bonn, Germany
External co-authors :
yes
Language :
English
Title :
Activation of the NLRP3 inflammasome in microglia: the role of ceramide.
ZonMw Internationale Stichting Alzheimer Onderzoek
Funding text :
The authors thank Francesco Santarelli for technical assistance, as well as Dr. Simone Crivelli for chemical compound exchange between the groups. This work was supported by the EU framework program JPND to MTH (InCure). PMM was supported by grants from ZonMw Memorabel program (projectnr: 733050105) and by the international foundation for Alzheimer Research (ISAO) (pro-jectnr: 14545). The authors declare that there is no conflict of interest with regard to the experimental part of this study. Michael T. Heneka is handling editor for the Journal of Neurochemistry.
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