title: Carbamoyl phosphate synthetase-1 is a rapid turnover biomarker in mouse and human acute liver injury.
abstract: Several serum markers are used to assess hepatocyte damage, but they have limitations related to etiology specificity and prognostication. Identification of novel hepatocyte-specific biomarkers could provide important prognostic information and better pathogenesis classification. We tested the hypothesis that hepatocyte-selective biomarkers are released after subjecting isolated mouse hepatocytes to Fas-ligand-mediated apoptosis. Proteomic analysis of hepatocyte culture medium identified the mitochondrial matrix protein carbamoyl phosphate synthetase-1 (CPS1) among the most readily detected proteins that are released by apoptotic hepatocytes. CPS1 was also detected in mouse serum upon acute challenge with Fas-ligand or acetaminophen and in hepatocytes upon hypoosmotic stress, independent of hepatocyte caspase activation. Furthermore, CPS1 was observed in sera of mice chronically fed the hepatotoxin 3,5-diethoxycarbonyl-1,4-dihydrocollidine. Mouse CPS1 detectability was similar in serum and plasma, and its half-life was 126 ± 9 min. Immune staining showed that CPS1 localized to mouse hepatocytes but not ductal cells. Analysis of a few serum samples from patients with acute liver failure (ALF) due to acetaminophen, Wilson disease, or ischemia showed readily detectable CPS1 that was not observed in several patients with chronic viral hepatitis or in control donors. Notably, CPS1 rapidly decreased to undetectable levels in sera of patients with acetaminophen-related ALF who ultimately recovered, while alanine aminotransferase levels remained elevated. Therefore, CPS1 becomes readily detectable upon hepatocyte apoptotic and necrotic death in culture or in vivo. Its abundance and short serum half-life, compared with alanine aminotransferase, suggest that it may be a useful prognostic biomarker in human and mouse liver injury.
authors.affiliation: Department of Molecular & Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan; sujith@umich.edu.
authors.forename: Sujith V W
authors.initials: SV
authors.lastname: Weerasinghe
authors.affiliation: Department of Molecular & Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan;
authors.forename: You-Jin
authors.initials: YJ
authors.lastname: Jang
authors.affiliation: Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan; and.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Fontana
authors.affiliation: Department of Molecular & Integrative Physiology, University of Michigan Medical School, Ann Arbor, Michigan; Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan; and VA Ann Arbor Healthcare System, Ann Arbor, Michigan.
authors.forename: M Bishr
authors.initials: MB
authors.lastname: Omary
mesh_terms: D000082:Acetaminophen; D000410:Alanine Transaminase / Q000097:blood; D000818:Animals; D017209:Apoptosis; D015415:Biomarkers / Q000378:metabolism; D002222:Carbamoyl-Phosphate Synthase (Ammonia) / Q000097:blood / Q000378:metabolism*; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000097:blood / Q000201:enzymology* / Q000209:etiology / Q000473:pathology; D056487:Chemical and Drug Induced Liver Injury, Chronic / Q000097:blood / Q000201:enzymology / Q000209:etiology; D017077:Culture Media, Conditioned / Q000378:metabolism; D004195:Disease Models, Animal; D053222:Fas Ligand Protein / Q000378:metabolism; D006207:Half-Life; D019694:Hepatitis B, Chronic / Q000097:blood / Q000201:enzymology; D019698:Hepatitis C, Chronic / Q000097:blood / Q000201:enzymology; D022781:Hepatocytes / Q000201:enzymology* / Q000473:pathology; D006801:Humans; D008099:Liver / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D009336:Necrosis; D011379:Prognosis; D011725:Pyridines; D013997:Time Factors
chemical_list: C530773:3,5-diethoxycarbonyl-1,4-dihydrocollidine; D015415:Biomarkers; D017077:Culture Media, Conditioned; D053222:Fas Ligand Protein; D011725:Pyridines; D000082:Acetaminophen; D000410:Alanine Transaminase; D002222:Carbamoyl-Phosphate Synthase (Ammonia)
keywords: HMGB1; acetaminophen; alanine aminotransferase; apoptosis; hepatocytes; necrosis