Metabolic syndrome, obesity, and steatosis are characterized by a range of dysregulations including defects in ubiquitin ligase tagging proteins for degradation. The identification of novel hepatic genes associated with fatty liver disease and metabolic dysregulation may be relevant to unravelling new mechanisms involved in liver disease progression METHODS: Through integrative analysis of liver transcriptomic and metabolomic obtained from obese subjects with steatosis, we identified itchy E ubiquitin protein ligase (ITCH) as a gene downregulated in human hepatic tissue in relation to steatosis grade. Wild-type or ITCH knockout mouse models of non-alcoholic fatty liver disease (NAFLD) and obesity-related hepatocellular carcinoma were analyzed to dissect the causal role of ITCH in steatosis RESULTS: We show that ITCH regulation of branched-chain amino acids (BCAAs) degradation enzymes is impaired in obese women with grade 3 compared with grade 0 steatosis, and that ITCH acts as a gatekeeper whose loss results in elevation of circulating BCAAs associated with hepatic steatosis. When ITCH expression was specifically restored in the liver of ITCH knockout mice, ACADSB mRNA and protein are restored, and BCAA levels are normalized both in liver and plasma CONCLUSIONS: Our data support a novel functional role for ITCH in the hepatic regulation of BCAA metabolism and suggest that targeting ITCH in a liver-specific manner might help delay the progression of metabolic hepatic diseases and insulin resistance.

Department of Biosciences Nottingham Trent University Nottingham NG11 8NS United Kingdom

Department of Diabetes Endocrinology and Nutrition University Hospital of Girona 'Dr Josep Trueta' Institut d'Investigacio Biomedica de Girona IdibGi; and CIBER Fisiopatologia de la Obesidad y Nutricion Girona Spain

Department of Diabetes Endocrinology and Nutrition University Hospital of Girona 'Dr Josep Trueta' Institut d'Investigacio Biomedica de Girona IdibGi; and CIBER Fisiopatologia de la Obesidad y Nutricion Girona Spain; Department of Medical Sciences School of Medicine University of Girona Spain

Department of Experimental Medicine University of Rome Tor Vergata Via Montpellier 1 00133 Rome Italy

Department of Surgery University of Rome Tor Vergata Via Montpellier 1 00133 Rome Italy

Department of Systems Medicine University of Rome Tor Vergata Via Montpellier 1 00133 Rome Italy

Department of Systems Medicine University of Rome Tor Vergata Via Montpellier 1 00133 Rome Italy; Center for Atherosclerosis University Hospital Policlinico Tor Vergata Italy

European Brain Research Institute Via del Fosso del Cavaliere 100 00131 Rome Italy

Imperial College London Section of Biomolecular Medicine Division of Systems Medicine Department of Metabolism Digestion and Reproduction Imperial College London Exhibition Road London SW7 2AZ United Kingdom

Imperial College London Section of Biomolecular Medicine Division of Systems Medicine Department of Metabolism Digestion and Reproduction Imperial College London Exhibition Road London SW7 2AZ United Kingdom; Section of Genomic and Environmental Medicine Respiratory Division National Heart and Lung Institute Imperial College London Dovehouse St London SW3 6LY United Kingdom; European Genomic Institute for Diabetes CNRS UMR 8199 INSERM UMR 1283 Institut Pasteur de Lille Lille University Hospital University of Lille 59045 Lille France; McGill University and Genome Quebec Innovation Centre 740 Doctor Penfield Avenue Montréal QC H3A 0G1 Canada

INSERM and University Paul Sabatier Institut des Maladies Métaboliques et Cardiovasculaires INSERM U1048 F 31432 Toulouse France and Université Paul Sabatier F 31432 Toulouse France

International Clinical Research Center London United Kingdom

Section of Human Anatomy Department of Biomedicine Neuroscience and Advanced Diagnostic Palermo Italy

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