Explore the role of hepatocyte inflammasome in alcoholic liver disease - Project Summary/Abstract Alcohol-Associated with are kidney regulated 1 all the inflammasome inflammasomes, essentially epithelial-targeted indicating revealed alcohol hand, hepatocytes r treatment. mitochondrial PPARα and cell the for features inhibiting beneficial Liver Disease (AALD) is the leading cause of liver-related deaths in the USA. AALD starts lipid deposition in hepatocytes and progresses to liver inflammation and fibrosis. Effective treatment options very limited. There are t hree clinical trials ongoing in humans targeting IL-1β signaling. In these studies, toxicity was reported, indicating that liver-specific approaches should be considered. IL-1β production is upstream by the assembly of inflammasome components [an inflammasome adaptor ASC, caspase- or caspase-11, and i nflammasome sensors (NLRs or NLRCs)]. nflammasome components are expressed in l iver cell types, and to date, we have very limited nowledge of the effect of inflammasomes in hepatocytes, most abundant liver cell type and an important driver of AALD pathophysiology. I t was reported that the was activated in hepatocytes upon alcohol exposure. To further explore the liver-specific role of we deleted one essential inflammasome component, caspase-1 (Casp1), common to all inflammasomes, in liver epithelial cells specifically using Albumin-Cre mice. We found that liver deletion of Casp1 leads to a strong exacerbation of the disease in the NIAAA alcohol model, a rotective role of Casp1 gainst AALD in the l iver. Liver bulk RNA-seq and proteomic analysis that in the absence of Casp1 from hepatocytes, the antioxidant defense mechanisms in mitochondria, detoxification, NF-κB-related pathways and myeloid-driven inflammation are reduced. On the other fatty acid synthesis and PPARα target genes are increased. In vitro experiments in Casp1-deficient evealed exacerbation of mitochondrial ROS production and cell death upon alcohol and LPS In Aim 1, we will examine which hepatocyte cell-intrinsic pathway is affected by Casp1 related to ROS production, alcohol metabolism, and lipid deposition. We will focus on NF-κB, β-catenin, and revealed by omics analysis. We will also examine if the Casp1 effect is due to IL-1β or IL-18 production signaling. In Aim 2, we will delete Casp1 or Casp11 in hepatocytes and examine liver pathology and immune alterations after 8 weeks of alcohol feeding. The results generated from this project will provide the basis for preparation of an R01 application to study which molecule from inflammasome sensors is the best candidate targeting in vivo in regular mice and in humanized mice with alcohol that we have een to recapitulate several of human AALD. These results will help in the design of better therapeutic approaches in humans by inflammasome function in the ell types where it has a clear pathogenic role while preserving the functions of the inflammasome. I k p a , s c
