title: Establishment and assessment of a new human embryonic stem cell-based biomarker assay for developmental toxicity screening.
abstract: A metabolic biomarker-based in vitro assay utilizing human embryonic stem (hES) cells was developed to identify the concentration of test compounds that perturbs cellular metabolism in a manner indicative of teratogenicity. This assay is designed to aid the early discovery-phase detection of potential human developmental toxicants. In this study, metabolomic data from hES cell culture media were used to assess potential biomarkers for development of a rapid in vitro teratogenicity assay. hES cells were treated with pharmaceuticals of known human teratogenicity at a concentration equivalent to their published human peak therapeutic plasma concentration. Two metabolite biomarkers (ornithine and cystine) were identified as indicators of developmental toxicity. A targeted exposure-based biomarker assay using these metabolites, along with a cytotoxicity endpoint, was then developed using a 9-point dose-response curve. The predictivity of the new assay was evaluated using a separate set of test compounds. To illustrate how the assay could be applied to compounds of unknown potential for developmental toxicity, an additional 10 compounds were evaluated that do not have data on human exposure during pregnancy, but have shown positive results in animal developmental toxicity studies. The new assay identified the potential developmental toxicants in the test set with 77% accuracy (57% sensitivity, 100% specificity). The assay had a high concordance (≥75%) with existing in vivo models, demonstrating that the new assay can predict the developmental toxicity potential of new compounds as part of discovery phase testing and provide a signal as to the likely outcome of required in vivo tests.
authors.affiliation: Stemina Biomarker Discovery, Inc, Madison, Wisconsin.
authors.forename: Jessica A
authors.initials: JA
authors.lastname: Palmer
authors.forename: Alan M
authors.initials: AM
authors.lastname: Smith
authors.forename: Laura A
authors.initials: LA
authors.lastname: Egnash
authors.forename: Kevin R
authors.initials: KR
authors.lastname: Conard
authors.forename: Paul R
authors.initials: PR
authors.lastname: West
authors.forename: Robert E
authors.initials: RE
authors.lastname: Burrier
authors.forename: Elizabeth L R
authors.initials: EL
authors.lastname: Donley
authors.forename: Fred R
authors.initials: FR
authors.lastname: Kirchner
mesh_terms: D001681:Biological Assay / Q000379:methods*; D015415:Biomarkers / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D047108:Embryonic Development / Q000187:drug effects; D053595:Embryonic Stem Cells / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D006801:Humans; D055432:Metabolomics; D008954:Models, Biological; D011247:Pregnancy; D013723:Teratogens / Q000633:toxicity; D018675:Toxicity Tests / Q000379:methods*
chemical_list: D015415:Biomarkers; D013723:Teratogens
keywords: biomarker identification; human embryonic stem cells; in vitro developmental toxicity screen; in vitro toxicology; mechanisms of teratogenesis; metabolomics; retinoic acid; valproic acid