Profiling of drugs and environmental chemicals for functional impairment of neural crest migration in a novel stem cell-based test battery

Abstract

Developmental toxicity in vitro assays have hitherto been established as stand-alone systems, based on a limited number of toxicants. Within the embryonic stem cell-based novel alternative tests project, we developed a test battery framework that allows inclusion of any developmental toxicity assay and that explores the responses of such test systems to a wide range of drug-like compounds. We selected 28 compounds, including several biologics (e.g., erythropoietin), classical pharmaceuticals (e.g., roflumilast) and also six environmental toxicants. The chemical, toxicological and clinical data of this screen library were compiled. In order to determine a non-cytotoxic concentration range, cytotoxicity data were obtained for all compounds from HEK293 cells and from murine embryonic stem cells. Moreover, an estimate of relevant exposures was provided by literature data mining. To evaluate feasibility of the suggested test framework, we selected a well-characterized assay that evaluates ‘migration inhibition of neural crest cells.’ Screening at the highest non-cytotoxic concentration resulted in 11 hits (e.g., geldanamycin, abiraterone, gefitinib, chlorpromazine, cyproconazole, arsenite). These were confirmed in concentration–response studies. Subsequent pharmacokinetic modeling indicated that triadimefon exerted its effects at concentrations relevant to the in vivo situation, and also interferon-β and polybrominated diphenyl ether showed effects within the same order of magnitude of concentrations that may be reached in humans. In conclusion, the test battery framework can identify compounds that disturb processes relevant for human development and therefore may represent developmental toxicants. The open structure of the strategy allows rich information to be generated on both the underlying library, and on any contributing assay.Electronic supplementary materialThe online version of this article (doi:10.1007/s00204-014-1231-9) contains supplementary material, which is available to authorized users.

Grouping of histone deacetylase inhibitors and other toxicants disturbing neural crest migration by transcriptional profiling

“…Cell migration was assessed with minor changes as described in Zimmer et al (2012Zimmer et al ( , 2014. On day 2 NCC were thawed and 50,000 cells/cm 2 were seeded in 48 well plates and were grown to a monolayer.…”

“…On this basis relevant assays could be combined into to a more efficient test battery (Piersma, 2014;Piersma et al, 2013;Zimmer et al, 2014).…”

Highlight report: acetaminophen hepatotoxicity

“…Currently, intensive research activities are ongoing to establish in vitro systems particularly for hepatotoxicity (Grinberg et al 2014;Ramaiahgari et al 2014;Rodrigues et al 2013;Schug et al 2013), nephrotoxicity (Sanchez-Niño et al 2014;Fujiki et al 2013;, neurotoxicity (Barbosa et al 2014;Sisnaiske et al 2014;Frimat et al 2010) and developmental toxicity (Krug et al 2013;Rempel et al 2015;Balmer et al 2014;Zimmer et al 2014). For validation of in vitro systems with hepatocytes APAP represents a popular reference compound (Jennings et al 2014;Hengstler et al 2014).…”

Highlight report: perspectives in stem cell research—unbiased quantification of the similarity between in vitro generated and primary hepatocytes

“…Particularly, in the fields of liver (Godoy et al 2013;Zellmer et al 2010;Godoy and Bolt 2012), kidney (Faiz et al 2015;Yang et al 2014), and neurotoxicity (Zimmer et al 2014;Waldmann et al 2014;Krug et al 2013), in vitro systems are frequently used. Primary human cells still represent a gold standard in toxicological research (Heise et al 2012;Ghallab 2013Ghallab , 2014a.…”