PROJECT TITLE: Role of macrophage evolution in hepatic adaptation to alcohol PROJECT SUMMARY / ABSTRACT Acute alcoholic hepatitis (AH) is a severe inflammatory liver disease triggered by binge drinking. The disease has a 30-day mortality of approximately 20%. One of the most striking features of AH is that it affects only a small minority of heavy drinkers suggesting that most individuals are protected from developing alcoholic liver disease by as yet unknown mechanisms. We have recently shown that alcohol exposure in mice causes early changes in liver macrophage (MΦ) populations with a loss of up to 50% of Kupffer cells and entry of infiltrating macrophages. By 10 days, Kupffer cell numbers are restored but their gene expression patterns have become more like anti-inflammatory M2 macrophages. By 35 days of alcohol exposure, the hepatic MΦ gene expression pattern changes further with a decrease in some of the classic M2 markers and the development of a gene expression pattern associated with a restorative MΦ phenotype. The macrophage population changes correlate with changes in the sensitivity of the liver to a challenge with LPS. We hypothesize that the “lost” Kupffer cells are rapidly replaced by “adaptive” macrophages leading to a state in which liver inflammation is minimal. This adaptive macrophage formation requires Kupffer cell-derived apoptotic bodies and Th2 cytokines such as IL4 and IL13. The balance of pro-and anti-inflammatory MΦs and the nature of the hepatic adaptive MΦ populations change over time so that with prolonged alcohol exposure, adaptation can be lost and liver inflammation can occur. Better understanding of the nature of the adaptive macrophages and the factors that lead to their formation, maintenance and loss would provide new approaches for the therapy of alcoholic liver disease. We will explore this hypothesis with the following specific aims: Aim 1: To determine the origins, gene expression patterns and functional properties of the mouse liver MΦ subtypes that appear after alcohol exposure. We will use lineage tracing techniques and single cell RNA sequencing to define the origin and diversity of the macrophage populations present. We will then isolate the adaptive macrophage populations to determine their functional properties both in vivo and in vitro. Aim 2: To define the signals responsible for production and maintenance of alcohol adaptive MΦ populations. We will examine the role of specific apoptotic body receptors, the impact of different sources of apoptotic bodies, timing of apoptotic body formation within the liver, and the role of hepatocyte derived factors in the formation of adaptive macrophage populations. Aim 3: To identify macrophage/monocyte populations that contribute to alcohol adaptation in humans. These experiments will leverage the mouse findings made in the first two aims to identify alcohol adaptive macrophage populations in humans. This will be done by immunohistochemical analysis of macropha...