You are here:
Expanding New Approach Methods for Developmental Toxicity: The DevTox Germ Layer Reporter Platform
Citation:
Gamble, J. AND C. Deisenroth. Expanding New Approach Methods for Developmental Toxicity: The DevTox Germ Layer Reporter Platform. Society of Toxicology 62nd Annual Meeting and ToxExpo, Nashville, TN, March 20 - 23, 2023. https://doi.org/10.23645/epacomptox.22695940
Impact/Purpose:
Poster to be presented at the Society of Toxicology 62nd Annual Meeting and ToxExpo March 2023.
Description:
The Frank R. Lautenberg Chemical Safety for the 21st Century Act mandates the US EPA develop new approach methods to detect chemical risks to susceptible populations, including pregnant women, that reduce reliance on the use of vertebrate animals in chemical testing. The DevTox-GLR (germ layer reporter) model platform was recently established for high-throughput screening and prioritization of potential developmental hazards (doi.org/10.3390/toxics10070392). The model platform utilizes the RUES2-GLR pluripotent stem cell reporter line that expresses fluorescent fusion protein biomarkers SOX17 (endoderm), Brachyury (mesoderm), and SOX2 (ectoderm and pluripotency); enabling a multi-lineage readout on gastrulation lineages. The endodermal reporter (DevTox GLR-Endo assay) employed use of the model platform to evaluate chemical effects on differentiating endoderm, yielding a balanced accuracy (BA) of 72% against a training set of 23 non-developmental toxicants and 43 developmental toxicants. To expand biological coverage, assays for pluripotency (DevTox GLR-Pluri), ectoderm (DevTox GLR-Ecto), and mesoderm (DevTox GLR-Meso) were developed to compare chemical-specific responses across lineages, thus providing a chemical specific mechanistic profile for perturbed gastrulation. Testing was done on each assay with a reference set of 4 non-developmental toxicants (acetaminophen, folic acid, penicillin G and saccharin) and 12 developmental toxicants (13-cis retinoic acid, 5,5-diphenylhydantoin, 5-fluouracil, all-trans retinoic acid, bisphenol A, busulfan, diethylstilbestrol, methotrexate, pomalidomide, sunitinib, thalidomide and valproic acid) with concentrations ranging from 100 pM to 200 µM. BAs were 96% (pluripotent), 83% (ectoderm), and 58% (mesoderm). Assay overlap revealed 7 chemicals correctly identified by all assays, although none of the tested chemicals had a lineage specific effect. However, developmental toxicity and cytotoxicity potencies varied across assays with differences between lineages. Further testing is necessary to determine the efficacy of each assay against a broader set of chemicals, but the DevTox GLR platform overall has the capability to determine mechanistic potencies of developmental toxicants on cell lineages specified during gastrulation. (This abstract does not necessarily reflect EPA policy, nor endorse or recommend any products mentioned).
URLs/Downloads:
DOI: Expanding New Approach Methods for Developmental Toxicity: The DevTox Germ Layer Reporter Platform
JOHNGAMBLE_SOT2023V5.PDF (PDF, NA pp, 863.478 KB, about PDF)