title: Identification of novel translational urinary biomarkers for acetaminophen-induced acute liver injury using proteomic profiling in mice.
abstract: Drug-induced liver injury (DILI) is the leading cause of acute liver failure. Currently, no adequate predictive biomarkers for DILI are available. This study describes a translational approach using proteomic profiling for the identification of urinary proteins related to acute liver injury induced by acetaminophen (APAP). Mice were given a single intraperitoneal dose of APAP (0-350 mg/kg bw) followed by 24 h urine collection. Doses of ≥275 mg/kg bw APAP resulted in hepatic centrilobular necrosis and significantly elevated plasma alanine aminotransferase (ALT) values (p<0.0001). Proteomic profiling resulted in the identification of 12 differentially excreted proteins in urine of mice with acute liver injury (p<0.001), including superoxide dismutase 1 (SOD1), carbonic anhydrase 3 (CA3) and calmodulin (CaM), as novel biomarkers for APAP-induced liver injury. Urinary levels of SOD1 and CA3 increased with rising plasma ALT levels, but urinary CaM was already present in mice treated with high dose of APAP without elevated plasma ALT levels. Importantly, we showed in human urine after APAP intoxication the presence of SOD1 and CA3, whereas both proteins were absent in control urine samples. Urinary concentrations of CaM were significantly increased and correlated well with plasma APAP concentrations (r = 0.97; p<0.0001) in human APAP intoxicants, who did not present with elevated plasma ALT levels. In conclusion, using this urinary proteomics approach we demonstrate CA3, SOD1 and, most importantly, CaM as potential human biomarkers for APAP-induced liver injury.
authors.affiliation: Department of Pharmacology and Toxicology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors.forename: Rachel P L
authors.initials: RP
authors.lastname: van Swelm
authors.forename: Coby M M
authors.initials: CM
authors.lastname: Laarakkers
authors.forename: Ellen C
authors.initials: EC
authors.lastname: van der Kuur
authors.forename: Eva
authors.initials: E
authors.lastname: Morava-Kozicz
authors.forename: Ron A
authors.initials: RA
authors.lastname: Wevers
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Augustijn
authors.forename: Daan J
authors.initials: DJ
authors.lastname: Touw
authors.forename: Maro H
authors.initials: MH
authors.lastname: Sandel
authors.forename: Rosalinde
authors.initials: R
authors.lastname: Masereeuw
authors.forename: Frans G M
authors.initials: FG
authors.lastname: Russel
mesh_terms: D000082:Acetaminophen / Q000009:adverse effects* / Q000494:pharmacology; D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D015415:Biomarkers / Q000652:urine; D002147:Calmodulin / Q000652:urine; D002256:Carbonic Anhydrases / Q000652:urine; D056486:Chemical and Drug Induced Liver Injury / Q000175:diagnosis / Q000378:metabolism* / Q000652:urine; D004305:Dose-Response Relationship, Drug; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D008875:Middle Aged; D020543:Proteome; D040901:Proteomics; D013482:Superoxide Dismutase / Q000652:urine; D000072105:Superoxide Dismutase-1; D055815:Young Adult
chemical_list: D015415:Biomarkers; D002147:Calmodulin; D020543:Proteome; C000606290:SOD1 protein, human; D000082:Acetaminophen; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D002256:Carbonic Anhydrases