title: Assessment of candidate biomarkers of drug-induced hepatobiliary injury in preclinical toxicity studies.
abstract: This study was designed to assess the value of a set of potential markers for improved detection of liver injury in preclinical toxicity studies. Male Wistar rats were treated with drug candidates (BAY16, EMD335823, BI-3) that previously failed during development, in part due to hepatotoxicity, at two dose levels for 1, 3 and 14 days. Concentrations of lipocalin-2/NGAL and clusterin, which are frequently overexpressed and released from damaged tissues, and thiostatin, recently identified within PredTox as being elevated in urine in response to liver injury, were determined in rat urine and serum by ELISA. This was supplemented by confirmatory qRT-PCR and immunohistochemical analyses in the target organ. Serum paraoxonase-1 activity (PON1), which has been suggested as a marker of hepatotoxicity, was determined using a fluorometric assay. Clusterin and PON1 were not consistently altered in response to liver injury. In contrast, thiostatin and NGAL were increased in serum and urine of treated animals in a time- and dose-dependent manner. These changes correlated well with mRNA expression in the target organ and generally reflected the onset and degree of drug-induced liver injury. Receiver-operating characteristics (ROC) analyses supported serum thiostatin, but not NGAL, as a better indicator of drug-induced hepatobiliary injury than conventional clinical chemistry parameters, i.e. ALP, ALT and AST. Although thiostatin, an acute phase protein expressed in a range of tissues, may not be specific for liver injury, our results indicate that thiostatin may serve as a sensitive, minimally-invasive diagnostic marker of inflammation and tissue damage in preclinical safety assessment.
authors.affiliation: University of Würzburg, Würzburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Adler
authors.forename: D
authors.initials: D
authors.lastname: Hoffmann
authors.forename: H
authors.initials: H
authors.lastname: Ellinger-Ziegelbauer
authors.forename: P
authors.initials: P
authors.lastname: Hewitt
authors.forename: K
authors.initials: K
authors.lastname: Matheis
authors.forename: L
authors.initials: L
authors.lastname: Mulrane
authors.forename: W M
authors.initials: WM
authors.lastname: Gallagher
authors.forename: J J
authors.initials: JJ
authors.lastname: Callanan
authors.forename: L
authors.initials: L
authors.lastname: Suter
authors.forename: M M
authors.initials: MM
authors.lastname: Fountoulakis
authors.forename: W
authors.initials: W
authors.lastname: Dekant
authors.forename: A
authors.initials: A
authors.lastname: Mally
mesh_terms: D000209:Acute-Phase Proteins / Q000378:metabolism; D000818:Animals; D043303:Aryldialkylphosphatase / Q000097:blood / Q000378:metabolism; D001659:Biliary Tract / Q000187:drug effects / Q000378:metabolism*; D054316:Biomarkers, Pharmacological / Q000097:blood / Q000378:metabolism* / Q000652:urine; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism*; D051152:Clusterin / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D022781:Hepatocytes / Q000187:drug effects; D007704:Kininogens / Q000097:blood / Q000378:metabolism / Q000652:urine; D000071068:Lipocalin-2; D054834:Lipocalins / Q000378:metabolism; D008297:Male; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D018675:Toxicity Tests
chemical_list: D000209:Acute-Phase Proteins; D054316:Biomarkers, Pharmacological; D051152:Clusterin; D007704:Kininogens; C489506:LCN2 protein, human; D000071068:Lipocalin-2; D054834:Lipocalins; D011518:Proto-Oncogene Proteins; D043303:Aryldialkylphosphatase