Project Summary Lipid accumulation, inflammation and metabolic dysregulation are hallmarks of liver disease. Nonalcoholic fatty liver disease(NAFLD) has recently emerged as one of the leading global etiologies of liver disease as it affects approximately 25% of the global population. Nonalcoholic steatohepatitis (NASH) is the inflammatory subtype of NAFLD, and approximately 40% of patients will progress to this stage; with another subset proceeding towards carcinogenesis. Unfortunately, many of these patients will go undetected as they progress to hepatocellular carcinoma which has a median survival of just 11 months. It is thus crucial to better characterize the interface between NAFLD and NASH to best improve patient outcomes. To better understand the molecular signals driving this transition we have targeted the insulin signaling pathway where we have established a liver specific deletion model of PTEN, the negative regulator of the insulin signaling pathway, that recapitulates human disease progression. Our preliminary data has shown that dysregulation in eicosanoid metabolism correlates with PTEN loss such that prostaglandin synthesis is significantly enhanced while proresolving cyp450 associated eicosanoid synthesis is downregulated. These correlations between PTEN, prostaglandin, and cyp450 associated protein expression levels are preserved agnostic of PTEN deletion when examining publicly available data. The PI3K/AKT signaling pathway is unequivocally induced upon PTEN deletion and as such we aim to further investigate the role of AKT in driving this bioactive lipid metabolism dysregulation. Utilizing genetic knockout hepatocytes, our data has shown that AKT regulates eicosanoid synthesis in an isoform specific manner. Further, eicosanoid have long been implicated in playing a role in modulating macrophage chemotaxis and polarization. Our data has also shown significant enrichment of macrophages, and previous work demonstrated that depletion of this macrophage accumulation via AKT2 deletion attenuated disease progression. As such, we aim to investigate the role of AKT specific isoforms in regulating hepatic eicosanoid biosynthesis, and the effect this has on macrophage chemotaxis and polarization in liver disease. Completion of this project will show the potential therapeutic benefit that targeting eicosanoid signaling may have in NAFLD and NASH. This work will also elucidate the mechanistic roles of each AKT isoform in regulating hepatic eicosanoid metabolism and chronic inflammation to provide more insights for the scientific community at large.