Detection of Drug-Induced Cholestasis Potential in Sandwich-Cultured Human Hepatocytes

Deferm, Neel; Richert, Lysiane; Brantegem, Pieter Van; Vocht, Tom De; Qi, Bing; Witte, Peter de; Bouillon, Thomas; Annaert, Pieter

doi:10.1007/978-1-4939-9420-5_22

Cited by 4 publications

(3 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The DICI reflects the ratio in urea production of SCHH co-exposed to a test compound and bile salts or to the test compound alone. In our previously published protocol [6] , [7] , [8] , compounds with a DICI value lower than 0.80 were considered cholestatic. This cut-off value was chosen arbitrarily and had no statistical basis.…”

Section: Discussionmentioning

confidence: 99%

“…By determining the ratio of urea formation after incubation with test compound in absence and in presence of the bile salt mixture, the DIC index (DICI) value can be calculated. Based on previously published protocols, a DICI value lower than 0.80 was used to flag a test compound at that specific concentration for an increased in vitro DIC risk [6] , [7] , [8] . In addition, following evaluation of a range of test compound concentrations, a safety margin (SM) could be estimated by dividing the lowest concentration with DICI < 0.80 (cf.…”

Section: General Description Of the Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Drug-induced cholestasis assay in primary hepatocytes

Brantegem

Chatterjee

Bruyn³

et al. 2020

MethodsX

Self Cite

View full text Add to dashboard Cite

Drug-induced cholestasis (DIC) is a major cause of clinical failure of drug candidates. Numerous patients worldwide are affected when exposed to marketed drugs exhibiting a DIC signature. Prospective identification of DIC during early compound development remains challenging. Here we describe the optimized in vitro procedure for early assessment and prediction of an increased DIC risk. Our method is based on three principles: Exposure of primary human hepatocyte cultures to test compounds in the absence and presence of a physiologically relevant mixture of endogenous bile salts. Rapid and quantitative assessment of the influence of concomitant bile salt exposure on hepatocyte functionality and integrity after 24 h or 48 h of incubation. Translation of the in vitro result, expressed as a DIC index (DICI) value, into an in vivo safety margin. Using our historical control data, a new (data driven) DICI cut-off value of 0.78 was established for discerning cholestatic and non-cholestatic compounds. Our DIC assay protocol was further improved by now relying on the principle of the no observable adverse effect level (NOAEL) for determining the highest test compound concentration corresponding to a DICI ≥ 0.78. Predicted safety margin values were subsequently calculated for compounds displaying hepatotoxic and/or cholestatic effects in patients, thus enabling evaluation of the performance of our DIC assay. Of note, this assay can be extended to explore the role of drug metabolites in precipitating DIC.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: General Description Of the Methodsmentioning

confidence: 99%

Drug-induced cholestasis assay in primary hepatocytes

Brantegem

Chatterjee

Bruyn³

et al. 2020

MethodsX

Self Cite

View full text Add to dashboard Cite

show abstract

“…The assay was able to adequately distinguish cholestatic compound from hepatotoxic non-cholestatic and non-hepatotoxic compounds. This SCH-based in vitro assay, thus, offers a unique tool to reliably detect cholestatic drug candidates Oorts et al 2016;Deferm et al 2019). In conclusion, the possibility to directly determine alterations in intracellular concentrations of BAs is one of the greatest assets of the SCH model, as it may provide useful mechanistic information on compounds that cause cholestasis ).…”

Section: Bei (%) =mentioning

confidence: 99%

Current insights in the complexities underlying drug-induced cholestasis

Deferm

Vocht

et al. 2019

Critical Reviews in Toxicology

Self Cite

View full text Add to dashboard Cite

Drug-induced cholestasis (DIC) poses a major challenge to the pharmaceutical industry and regulatory agencies. It causes both drug attrition and post-approval withdrawal of drugs. DIC represents itself as an impaired secretion and flow of bile, leading to the pathological hepatic and/or systemic accumulation of bile acids (BAs) and their conjugate bile salts. Due to the high number of mechanisms underlying DIC, predicting a compound's cholestatic potential during early stages of drug development remains elusive. A profound understanding of the different molecular mechanisms of DIC is, therefore, of utmost importance. Although many knowledge gaps and caveats still exist, it is generally accepted that alterations of certain hepatobiliary membrane transporters and changes in hepatocellular morphology may cause DIC. Consequently, liver models, which represent most of these mechanisms, are valuable tools to predict human DIC. Some of these models, such as membrane-based in vitro models, are exceptionally well-suited to investigate specific mechanisms (i.e., transporter inhibition) of DIC, while others, such as liver slices, encompass all relevant biological processes and therefore offer a better representation of the in vivo situation. In the current review, we highlight the principal molecular mechanisms associated with DIC and offer an overview and critical appraisal of the different liver models that are currently being used to predict the cholestatic potential of drugs.

show abstract

Primary hepatocyte cultures for liver disease modeling

Vinken¹

2021

Current Opinion in Toxicology

View full text Add to dashboard Cite

show abstract

Detection of Drug-Induced Cholestasis Potential in Sandwich-Cultured Human Hepatocytes

Cited by 4 publications

References 17 publications

Drug-induced cholestasis assay in primary hepatocytes

Drug-induced cholestasis assay in primary hepatocytes

Current insights in the complexities underlying drug-induced cholestasis

Primary hepatocyte cultures for liver disease modeling

Contact Info

Product

Resources

About