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

Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Toxicology and Applied Pharmacology (Impact Factor: 3.71). 05/2009; 237(3):317-30. DOI: 10.1016/j.taap.2009.04.002
Source: PubMed


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, IFN gamma, IL-1 alpha, 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-1 alpha, 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.

Download full-text


Available from: Bruce Tidor,
40 Reads
  • Source
    • "3) Probes: Expand fluorescent imaging probes to include additional bile acid analogs that are substrates of both NTCP and BSEP transporters, in addition to cholyl lysyl fluoresceine which is a more specific substrate for OATP/MRP transporters (de Waart et al., 2010). 4) Inflammatory and multi-cell systems: Explore the role of pro-inflammatory cytokines and/or Kupffer cells for possible synergistic effects in test sensitivity without sacrificing specificity (Cosgrove et al., 2009), and assess the potential benefit of 3D liver chip with controlled microfluidics. 5) Modeling: Mechanism-based PKPD modeling approaches should be applied to integrate any in vitro measurements into a holistic in vivo prediction (Woodhead et al., 2014). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Incorporating phenotypic screening as a key strategy enhances predictivity and translatability of drug discovery efforts. Cellular imaging serves as a "phenotypic anchor" to identify important toxicologic pathology that encompasses an array of underlying mechanisms, thus provides an effective means to reduce drug development failures due to insufficient safety. This mini-review highlights the latest advances in hepatotoxicity, cardiotoxicity, and genetic toxicity tests that utilized cellular imaging as a screening strategy, and recommends path forward for further improvement.
    Frontiers in Pharmacology 10/2015; 6:191. DOI:10.3389/fphar.2015.00191 · 3.80 Impact Factor
  • Source
    • "The choice of Dicrocoelium as model for the present study is related to both the poor knowledge on its pathogenesis, very often masked by the frequent poliparasitism in ruminants, and to its high prevalence in sheep, goats, cattle and buffaloes of southern Italy (Cringoli et al., 2002; Maurelli et al., 2007; Rinaldi et al., 2009; Musella et al., 2011) and other regions of the Mediterranea area (Rojo-Vázquez et al., 2012). Specifically, in the present study we investigated the effects of somatic antigen of D. dendriticum on cell death mechanisms using two human hepatocyte cell lines, HepG2 and HuH7, representing an almost non-tumor (Cosgrove et al., 2009; Soldatow et al., 2013) and a tumor cell line, respectively. "
    [Show abstract] [Hide abstract]
    ABSTRACT: The relationship between Dicrocoelium dendriticum and cancer has been poorly investigated so far, but a large amount of findings suggest that other trematodes can favour cancer in both animals and humans. In this study, the effects of D. dendriticum on cell proliferation, cell death mechanisms and oxidative stress induction were evaluated in hepatocellular carcinoma (HCC) cell lines (HepG2 and HuH7). Results showed that short time exposure to low concentrations of somatic antigens from D. dendriticum caused slight proliferation in both HepG2 and HuH7 cells while high concentrations and long exposure time to extracts from D. dendriticum caused a significant growth inhibition. This effect was, however, not paralleled by apoptosis but it occurred with an about 40% increase of the formation of autophagic vacuoles. In the same experimental conditions, a strong oxidative stress was recorded with an about 100% increase of the intracellular O2-. These data suggest the occurrence of an escape anti-apoptotic mechanism in HCC cells. In conclusion, these results suggest a role for D. dendriticum in the chronic oxidative stress and in the regulation of transformation processes in HCC warranting additional investigations in this specific area of research.
    Veterinary Parasitology 08/2015; 212(3). DOI:10.1016/j.vetpar.2015.07.039 · 2.46 Impact Factor
    • "The potential utility of this new assay to screen metabolism-dependent hepatotoxins was evidenced by a set of bioactivable and nonbioactivable drugs (Tolosa et al. 2013). Similarly, several authors have proposed HCS-based assays to explore drug toxicity to a variety of liver cell models, including rodent hepatocytes (Chen et al. 2014; Cosgrove et al. 2009), hepatocytes cocultured with fibroblasts (Cole et al. 2014), HepaRG cells (Pernelle et al. 2011; Ranade et al. 2014), or HepG2 cells maintained in microfluidic devices (Ye et al. 2007) or combined with subcellular liver fractions (Garside et al. 2014; Westerink et al. 2011). These multiparametric strategies offer the possibility of detecting subtle toxicity-related changes with greater sensitivity than monoparametric cytotoxicity assays which detect overt toxicity (O'Brien et al. 2006; Tolosa et al. 2012b). "
    [Show abstract] [Hide abstract]
    ABSTRACT: High-content screening is the application of automated microscopy and image analysis to both cell biology and drug discovery. Over the last decade, this technique has emerged as a useful technology that allows the simultaneous measurement of different parameters at a single-cell level. Hepatotoxicity is a compelling reason for drug nonapprovals and withdrawals. It is recognized that the safety of a compound cannot be based on a single in vitro assay, and existing methods are not predictive of drug-induced toxicity. However, different HCS assays have been recently demonstrated as being powerful for identifying different mechanisms implicated in drug-induced toxicity with high sensitivity and specificity. These assays integrate the data obtained from different cell function indicators and can be easily incorporated into basic screening processes for the safety evaluation and selection of drug candidates; thus, they contribute greatly to lessen the likelihood of drug failure. Exploring the use of cellular imaging technology in drug-induced liver injury by reviewing the different tests proposed provides evidence that this technology has a strong impact on drug discovery.
    Archives of Toxicology 03/2015; 89(7). DOI:10.1007/s00204-015-1503-z · 5.98 Impact Factor
Show more