Vesicle- and Hepatocyte-Based Assays for Identification of Drug Candidates Inhibiting BSEP Function

Brantegem, Pieter Van; Deferm, Neel; Qi, Bing; Vocht, Tom De; Annaert, Pieter

doi:10.1007/978-1-4939-9420-5_4

Cited by 2 publications

(1 citation statement)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Potential interactions of new molecular entities with BSEP and MRP3/MRP4 have therefore likely to be considered during their pharmaceutical development, in order to identify and discard candidate drugs that may cause DILIs (Hillgren et al., 2013 ; Kenna et al., 2018 ; Morgan et al., 2010 , 2013 ). For such a purpose, various in vitro assays are available, such as sandwich‐cultured hepatocytes‐based efflux assays and BSEP‐, MRP3‐, or MRP4‐related vesicular assays (Cheng et al., 2016 ; Kenna et al., 2018 ; Kostrubsky et al., 2003 ; Pedersen et al., 2013 ; Van Brantegem et al., 2019 ; Wolf et al., 2010 ). The use of cultured hepatocytes, but not that of vesicles, allows an integrated approach, with determination of drug effects towards both canalicular and sinusoidal efflux of bile acids such as taurocholate (TC) (Susukida et al., 2015 ).…”

Section: Introductionmentioning

confidence: 99%

Inhibition of canalicular and sinusoidal taurocholate efflux by cholestatic drugs in human hepatoma HepaRG cells

Vée

Moreau

Jouan

et al. 2022

Biopharm & Drug Disp

View full text Add to dashboard Cite

HepaRG cells are highly-differentiated human hepatoma cells, which are increasingly recognized as a convenient cellular model for in vitro evaluation of hepatic metabolism, transport, and/or toxicity of drugs. The present study was designed to evaluate whether HepaRG cells can also be useful for studying drug-mediated inhibition of canalicular and/or sinusoidal hepatic efflux of bile acids, which constitutes a major mechanism of drug-induced liver toxicity. For this purpose, HepaRG cells, initially loaded with the bile acid taurocholate (TC), were reincubated in TC-free transport assay medium, in the presence or absence of calcium or drugs, before analysis of TC retention. This method allowed us to objectivize and quantitatively measure biliary and sinusoidal efflux of TC from HepaRG cells, through distinguishing cellular and canalicular compartments. In particular, time-course analysis of the TCfree reincubation period of HepaRG cells, that is, the efflux period, indicated that a 20 min-efflux period allowed reaching biliary and sinusoidal excretion indexes for TC around 80% and 60%, respectively. Addition of the prototypical cholestatic drugs bosentan, cyclosporin A, glibenclamide, or troglitazone during the TC-free efflux phase period was demonstrated to markedly inhibit canalicular and sinusoidal secretion of TC, whereas, by contrast, incubation with the noncholestatic compounds salicylic acid or flumazenil was without effect. Such data therefore support the use of human HepaRG cells for in vitro predicting drug-induced liver toxicity (DILI) due to the inhibition of hepatic bile acid secretion, using a biphasic TC loading/efflux assay. K E Y W O R D Sbile salt export pump, cholestatic drug, efflux, HepaRG cells, taurocholate | INTRODUCTIONAlteration of hepatic bile acid transport is recognized as a major mechanism of drug-induced liver injury (drug-induced liver toxicity [DILI]) (Chatterjee & Annaert, 2018;Fernández-Murga et al., 2018;Mosedale & Watkins, 2017). It notably corresponds to the inhibition of the canalicular ATP-binding cassette (ABC) transporter bile salt export pump (BSEP/ABCB11) and results in bile acid retention into hepatocytes and cholestasis (Dawson et al., 2012;Izat & Sahin, 2021). Besides canalicular efflux, sinusoidal secretion of bile acids, mediatedThis is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

show abstract