Comparisons betweenin vitrowhole cell imaging andin vivozebrafish-based approaches for identifying potential human hepatotoxicants earlier in pharmaceutical development

Abstract

Drug-induced liver injury (DILI) is a major cause of attrition during both the early and later stages of the drug development and marketing process. Reducing or eliminating drug-induced severe liver injury, especially those that lead to liver transplants or death, would be tremendously beneficial for patients. Therefore, developing new pharmaceuticals that have the highest margins and attributes of hepatic safety would be a great accomplishment. Given the current low productivity of pharmaceutical companies and the high costs of bringing new medicines to market, any early screening assay(s) to identify and eliminate pharmaceuticals with the potential to cause severe liver injury in humans would be of economic value as well. The present review discusses the background, proof-of-concept, and validation studies associated with high-content screening (HCS) by two major pharmaceutical companies (Pfizer Inc and Jansen Pharmaceutical Companies of Johnson & Johnson) for detecting compounds with the potential to cause human DILI. These HCS assays use fluorescent-based markers of cell injury in either human hepatocytes or HepG2 cells. In collaboration with Evotec, an independent contract lab, these two companies also independently evaluated larval zebrafish as an early-stage in vivo screen for hepatotoxicity in independently conducted, blinded assessments. Details about this model species, the need for bioanalysis, and, specifically, the outcome of the phenotypic-based zebrafish screens are presented. Comparing outcomes in zebrafish against both HCS assays suggests an enhanced detection for hepatotoxicants of most DILI concern when used in combination with each other, based on the U.S. Food and Drug Administration DILI classification list.

Graphene-Based Anticancer Nanosystem and Its Biosafety Evaluation Using a Zebrafish Model

“…[30][31][32]62,67 Compared with the blank control, neither sublethal toxicity nor drop of hatching rate was detectable in the presence of lGO/NPcurcumin complexes. Screening of phenotypic toxicity may be implemented because of opaque quality of tissues following cell death.…”

“…33,62 Noteworthy, a short-term zebrafish biotoxicity assessment could be more or less linked to the developmental biotoxicity during the long-term human growth. 32,63 Herein, we investigated the biotoxicity of graphene-based nanomaterials using zebrafish as a vertebrate model from the embryo through the hatchling stage. 62,64 Water-insoluble curcumin is a potentially anticancer drug, stemming from its ability to modulate growth of tumor cells through regulation of multiple cell signaling pathways.…”

Liver Fatty Acid Binding Protein Deficiency Provokes Oxidative Stress, Inflammation, and Apoptosis-Mediated Hepatotoxicity Induced by Pyrazinamide in Zebrafish Larvae

“…Their high fecundity and rapid organogenesis, the transparent nature of embryos and larvae, the ability to screen the whole organism in a microtiter plate format with small amounts (single milligrams) of compounds, and especially their high degree of genetic conservation with humans offer additional advantages over traditional in vivo animal models (11). As a vertebrate, high-level genetic conservation with mammals in liver development, regeneration, and carcinogenesis was documented (12), and a good correlation with mammalian hepatotoxicity has been observed (13).…”

Acute hepatotoxicity induced by quetiapine fumarate in larval zebrafish

“…The mor-phological and molecular basis of tissues and organs in zebrafish is either identical or similar to humans, possessing orthologs for ~85% of human genes and > 86% of human drug targets (Chen and Fishman, 1996;Granato and Nüsslein-Volhard, 1996;Gunnarsson et al, 2008;Howe et al, 2013). It has been confi rmed that zebrafi sh has a strikingly similar toxicity profi le to mammalian and is more appropriate for identifying endpoints of toxicity and elucidating the toxicity mechanisms (McGrath and Li, 2008;Hill et al, 2012;Hill, 2005). Zebrafish complete primary liver morphogenesis by 48 hr post-fertilization (hpf) and the liver is fully formed and function by 72 hpf.…”