Synergistic drug–cytokine induction of hepatocellular death as an in vitro approach for the study of inflammation-associated idiosyncratic drug hepatotoxicity

Abstract

Idiosyncratic drug hepatotoxicity represents a major problem in drug development due to inadequacy of current preclinical screening assays, but recently established rodent models utilizing bacterial LPS co-administration to induce an inflammatory background have successfully reproduced idiosyncratic hepatotoxicity signatures for certain drugs. However, the low-throughput nature of these models renders them problematic for employment as preclinical screening assays. Here, we present an analogous, but high-throughput, in vitro approach in which drugs are administered to a variety of cell types (primary human and rat hepatocytes and the human HepG2 cell line) across a landscape of inflammatory contexts containing LPS and cytokines TNF, IFNγ, IL-1α, and IL-6. Using this assay, we observed drug-cytokine hepatotoxicity synergies for multiple idiosyncratic hepatotoxicants (ranitidine, trovafloxacin, nefazodone, nimesulide, clarithromycin, and telithromycin) but not for their corresponding non-toxic control compounds (famotidine, levofloxacin, buspirone, and aspirin). A larger compendium of drug-cytokine mix hepatotoxicity data demonstrated that hepatotoxicity synergies were largely potentiated by TNF, IL-1α, and LPS within the context of multi-cytokine mixes. Then, we screened 90 drugs for cytokine synergy in human hepatocytes and found that a significantly larger fraction of the idiosyncratic hepatotoxicants (19%) synergized with a single cytokine mix than did the non-hepatotoxic drugs (3%). Finally, we used an information theoretic approach to ascertain especially informative subsets of cytokine treatments for most highly effective construction of regression models for drug- and cytokine mix-induced hepatotoxicities across these cell systems. Our results suggest that this drug-cytokine co-treatment approach could provide a useful preclinical tool for investigating inflammation-associated idiosyncratic drug hepatotoxicity.

A testing strategy to predict risk for drug-induced liver injury in humans using high-content screen assays and the ‘rule-of-two’ model

“…Notably, in the study of Cosgrove et al (2009), human hepatocyte cultures were treated with a mix of individual drugs and different cytokines. Approximately 25 % sensitivity and 97 % specificity for DILI prediction were reported using a large compendium of drugs.…”

“…Approximately 25 % sensitivity and 97 % specificity for DILI prediction were reported using a large compendium of drugs. The data from the in vitro assay of Cosgrove et al (2009) were combined with the RO2 model using the proposed testing strategy for an improved prediction of DILI (Supplemental Table 2). …”

“…For this purpose, a primary rat hepatocyte-based HCS assay was studied by investigating the negatives predicted from the RO2 model, thereby refining the results obtained from in silico methods alone. Furthermore, we implemented published data on drug-induced cytokine release (Cosgrove et al 2009) into our testing strategy to improve its predictive performance. Our study demonstrates that the proposed testing strategy improves the accuracies of detecting risk for DILI by approximately 10 % and at the same time contributes to the reduction of experimental cost by assisting in the rational selection of candidates for HCS and subsequent animal studies.…”

Mitochondrial and immunoallergic injury increase risk of positive drug rechallenge after drug-induced liver injury: A systematic review

“…1,[48][49][50][51] Most drugs resulting in positive rechallenge are administered at a high daily drug dose (>50 mg), which has been associated with a higher risk of DILI overall. 52 Typically, fewer than 1 in 1,000 exposed patients develop severe DILI, 53 suggesting a heightened vulnerability in those affected, which may be due to a concurrent, potentially transient, inflammation 54,55 and resultant oxidative stress, with concomitant medications contributing to defective liver regeneration/ repair, 56 high drug dose or hepatic metabolism, 52 female sex or obesity, 28 inherited pathogenic mitochondrial DNA mutations, 12 other genetic susceptibility, 57 or other factors. Therefore, in addition to drugspecific mechanisms of injury, interindividual factors greatly influence the reaction to DILI.…”

Idiosyncratiques et « omiques » peuvent-ils rimer ?

“…Une étude récente a consisté à stimuler des cellules hépatiques (hépatocytes primaires humains, hépato-cytes de rats, lignée d'hépatome humain HepG2) par un cocktail cytokinique (IFN , TNF, IL-1, IL-6). Une cytotoxicité a été ensuite observée lors d'un traitement par des médicaments induisant des réactions idiosyncratiques (ranitidine, TVX, nimésulide), mais non lors d'un traitement par des médicaments non hépatotoxiques (famotidine, LVX, aspirine) [27].…”