Inhibition of bile canalicular network formation in rat sandwich cultured hepatocytes by drugs associated with risk of severe liver injury. (September 2016)

• Record Type: Journal Article
• Authors: Takemura, Akinori; Izaki, Aya; Sekine, Shuichi; Ito, Kousei
• Is Part Of: Toxicology in vitro. Volume 35(2016)
• Page Start: 121
• Page End: 130
• Publication Date: 2016-09
• Subjects: DILI drug-induced liver injury -- ALT alanine aminotransferase -- ALP alkaline phosphatase -- BBW black box warning -- WDN withdrawn -- BC bile canalicular -- BSEP bile salt export pump -- MRP2 multidrug resistance-associated protein 2 -- SCHs sandwich cultured hepatocytes -- WME Williams' Medium E -- DMEM Dulbecco's Modified Eagle's Medium -- TCA taurocholic acid -- LCA lithocholic acid -- LDH lactate dehydrogenase -- IC50 half maximal inhibitory concentration -- FDA Food and Drug Administration -- MDD maximum daily dose -- PMDA Pharmaceuticals and Medical Devices Agency -- Cmax, u maximum unbound concentration in plasma -- NM no mention -- AR adverse reaction -- WNG warning -- PC precaution -- EC50 half maximal effective concentration; Bile canaliculus -- Sandwich cultured hepatocytes -- Hepatotoxicity -- In vitro screening -- Cholestasis -- Jaundice; Toxicity testing -- In vitro -- Periodicals; Toxicology -- Periodicals; 615.9
• DOI: 10.1016/j.tiv.2016.05.016
• Languages: English
• ISSNs: 0887-2333

Abstract

Abstract: Idiosyncratic drug-induced liver injury is a clinical concern with serious consequences. Although many preclinical screening methods have been proposed, it remains difficult to identify compounds associated with this rare but potentially fatal liver condition. Here, we propose a novel assay system to assess the risk of liver injury. Rat primary hepatocytes were cultured in a sandwich configuration, which enables the formation of a typical bile canalicular network. From day 2 to 3, test drugs, mostly selected from a list of cholestatic drugs, were administered, and the length of the network was semi-quantitatively measured by immunofluorescence. Liver injury risk information was collected from drug labels and was compared with in vitro measurements. Of 23 test drugs examined, 15 exhibited potent inhibition of bile canalicular network formation (< 60% of control). Effects on cell viability were negligible or minimal as confirmed by lactate dehydrogenase leakage and cellular ATP content assays. For the potent 15 drugs, IC50 values were determined. Finally, maximum daily dose divided by the inhibition constant gave good separation of the highest risk of severe liver toxicity drugs such as troglitazone, benzbromarone, flutamide, and amiodarone from lower risk drugs. In conclusion, inhibitory effect on the bile canalicular network formation observed in in vitro sandwich cultured hepatocytes evaluates a new aspect of drug toxicity, particularly associated with aggravation of liver injury. Graphical abstract: Highlights: The bile canalicular network is a prerequisite for biliary excretion. Bile canalicular network formation was quantified in vitro . Some drugs potently inhibited bile canalicular network formation. Inhibition by drugs correlated with the severity of in vivo liver toxicity. Our novel assay system is useful to predict the risk of drug-induced liver injury.