Potential conflict of interest: Dr. Hoffman is a consultant for Sobi, Regeneron, and Novartis. He advises, is on the speakers' bureau for, and received grants from Novartis and Sobi.
Steatohepatitis/Metabolic Liver Disease
NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice
Article first published online: 30 JAN 2014
© 2014 by the American Association for the Study of Liver Diseases
Volume 59, Issue 3, pages 898–910, March 2014
How to Cite
Wree, A., Eguchi, A., McGeough, M. D., Pena, C. A., Johnson, C. D., Canbay, A., Hoffman, H. M. and Feldstein, A. E. (2014), NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice. Hepatology, 59: 898–910. doi: 10.1002/hep.26592
The work was funded by the National Institutes of Health (DK076852 and DK082451 [to A.E.F.] and AI52430 [to H.M.H.]) and the German Research Foundation (DFG grant no. 173/2-1; to A.W.). The monoclonal alpha-tubulin antibody, developed by J. Frankel and E.M. Nelsen, was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the Eunice Kennedy Shriver National Institute of Child Health and Human Development and maintained by the Department of Biology at The University of Iowa (Iowa City, IA).
See Editorial on Page 761
- Issue published online: 25 FEB 2014
- Article first published online: 30 JAN 2014
- Accepted manuscript online: 28 JUN 2013 02:29PM EST
- Manuscript Accepted: 14 JUN 2013
- Manuscript Received: 8 JAN 2013
Inflammasome activation plays a central role in the development of drug-induced and obesity-associated liver disease. However, the sources and mechanisms of inflammasome-mediated liver damage remain poorly understood. Our aim was to investigate the effect of NLRP3 inflammasome activation on the liver using novel mouse models. We generated global and myeloid cell-specific conditional mutant Nlrp3 knock-in mice expressing the D301N Nlrp3 mutation (ortholog of D303N in human NLRP3), resulting in a hyperactive NLRP3. To study the presence and significance of NLRP3-initiated pyroptotic cell death, we separated hepatocytes from nonparenchymal cells and developed a novel flow-cytometry–based (fluorescence-activated cell sorting; FACS) strategy to detect and quantify pyroptosis in vivo based on detection of active caspase 1 (Casp1)- and propidium iodide (PI)-positive cells. Liver inflammation was quantified histologically by FACS and gene expression analysis. Liver fibrosis was assessed by Sirius Red staining and quantitative polymerase chain reaction for markers of hepatic stellate cell (HSC) activation. NLRP3 activation resulted in shortened survival, poor growth, and severe liver inflammation; characterized by neutrophilic infiltration and HSC activation with collagen deposition in the liver. These changes were partially attenuated by treatment with anakinra, an interleukin-1 receptor antagonist. Notably, hepatocytes from global Nlrp3-mutant mice showed marked hepatocyte pyroptotic cell death, with more than a 5-fold increase in active Casp1/PI double-positive cells. Myeloid cell-restricted mutant NLRP3 activation resulted in a less-severe liver phenotype in the absence of detectable pyroptotic hepatocyte cell death. Conclusions: Our data demonstrate that global and, to a lesser extent, myeloid-specific NLRP3 inflammasome activation results in severe liver inflammation and fibrosis while identifying hepatocyte pyroptotic cell death as a novel mechanism of NLRP3-mediated liver damage. (Hepatology 2014;59:898–910)