Project Summary/Abstract We aim to comprehensively determine how alcohol consumption alters the metabolism of critical organs, which can contribute to alcohol-induced organ damage. Alcohol consumption is the leading cause of public health burden worldwide. Chronic alcohol consumption is tightly associated with the development of cirrhosis, kidney dysfunction, intestinal inflammation, immune dysregulation, neurological damage, and cancers. However, it is still incompletely understood how alcohol and its metabolic products impact organ metabolism and potentially cause organ damage. Metabolism is a dynamic process like a flowing river. However, conventional methods that measure static snapshots such as gene expression, protein, or metabolite levels do not provide information about metabolic activities. For example, increased blood glucose levels can be due to either elevated production or reduced consumption by a dozen of organs. To resolve this issue, we will employ our unique platform, arteriovenous (AV) metabolomics, to quantitatively measure the dynamic metabolic activities of all organs relevant to alcoholic diseases (Aim 1). We will collect arterial blood and 10 organ-specific venous blood from alcohol-fed subjects and measure metabolite concentration gradients using liquid- chromatography mass spectrometry (LC-MS). This will inform how much and what kind of metabolites are absorbed or released by each organ. For this study, we will use pigs because pigs are similar to humans in terms of voluntary alcohol drinking behavior, addiction/intoxication phenotypes, and alcohol metabolic rate. By measuring ~1,300 metabolites’ movements between 10 major organs in pigs after alcohol feeding for three different durations, we will elucidate how each organ’s metabolism changes by alcohol consumption over time. To identify genes that drive alcohol-induced organ metabolic changes, we will also perform RNA-seq in the same pig tissues and employ new bioinformatics analysis to integrate these two omics data (Aim 2). Dr. Marcus Seldin, a bioinformatics expert, will then perform a battery of systematic correlation analyses using his innovative computational tools. Finally, we will validate the top candidate gene-metabolite relationships in cultured cells using gain/loss-of-function experiments. Such genes that mediate the alcohol-induced organ metabolism changes can be drug targets to mitigate alcohol’s detrimental effects. Our study will thus delineate previously unrecognized metabolic changes by alcohol in multiple organs as well as the key responsible genes, illuminating the way for targetable therapy to prevent alcohol-induced organ metabolic remodeling.