Sandwich-Cultured Hepatocytes as a Tool to Study Drug Disposition and Drug-Induced Liver Injury

Abstract

Sandwich-cultured hepatocytes (SCH) are metabolically competent and have proper localization of basolateral and canalicular transporters with functional bile networks. Therefore, this cellular model is a unique tool that can be used to estimate biliary excretion of compounds. SCH have been used widely to assess hepatobiliary disposition of endogenous and exogenous compounds and metabolites. Mechanistic modeling based on SCH data enables estimation of metabolic and transporter-mediated clearances, which can be employed to construct physiologically-based pharmacokinetic models for prediction of drug disposition and drug-drug interactions in humans. In addition to pharmacokinetic studies, SCH also have been employed to study cytotoxicity and perturbation of biological processes by drugs and hepatically-generated metabolites. Human SCH can provide mechanistic insights underlying clinical drug-induced liver injury (DILI). In addition, data generated in SCH can be integrated into systems pharmacology models to predict potential DILI in humans. In this review, applications of SCH in studying hepatobiliary drug disposition and bile acid-mediated DILI are discussed. An example is presented to show how data generated in the SCH model was used to establish a quantitative relationship between intracellular bile acids and cytotoxicity, and how this information was incorporated into a systems pharmacology model for DILI prediction.

Effect of Liver Disease on Hepatic Transporter Expression and Function

“…For example, MRP3-6 are induced in liver biopsies from NASH patients, similar to findings in a rodent model of NASH. 176,180–182 Similarly, Mrp1 mRNA is increased ~5-fold compared to control animals. 176 Functional drug disposition studies corroborate these reports as evidenced by increased plasma acetaminophen glucuronide concentrations in diet-induced NASH animals attributable to increased Mrp3 expression.…”

Farnesoid X Receptor Agonists Obeticholic Acid and Chenodeoxycholic Acid Increase Bile Acid Efflux in Sandwich-Cultured Human Hepatocytes: Functional Evidence and Mechanisms

“…Model-independent parameters describing the overall hepatobiliary disposition of TCA in SCHH were calculated using eqs. 1 and 2 (Liu et al, 1999;Yang et al, 2016) as well as eq. 3.…”

“…(Yu et al, 2002;Modica et al, 2010), multidrug resistanceassociated protein 2 (MRP2) (Modica et al, 2010;Liu et al, 2014), and organic solute transporter (OSTa/b) (Boyer et al, 2006;Modica et al, 2010;Liu et al, 2014) in hepatic cell lines and primary human hepatocytes. Sandwich-cultured human hepatocytes (SCHH) are a physiologically relevant model that maintains transporter function, morphology, and regulatory machinery (LeCluyse et al, 2000;Yang et al, 2016) and therefore, is an ideal system to study the regulation of bile acid transporters after nonacute (.24-hour) treatment of hepatocytes with FXR agonists. Recent studies revealed that OCA and CDCA treatment of SCHH for 72 hours increased mRNA expression of OSTa/b and BSEP with minor changes in MRP3, MRP4, and sodium taurocholate cotransporting polypeptide (NTCP) (Jackson et al, 2016;Zhang et al, 2017).…”

Characterization of hepatocyte-based in vitro systems for reliable toxicity testing

“…Of more pharmaco-toxicological relevance are drug transporter assays, which offer the advantage to combine testing of polarity and hepatocellular functionality. In this respect, functionality testing of the sinusoidal sodium-taurocholate cotransporter, the apical bile salt export pump and multidrug resistance protein 2, using fluorescent probes and inhibitors, is a possible suggestion (Ramboer et al 2013;Yang et al 2016). …”