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Modeling Endodermal Differentiation Trajectories Following All-trans Retinoic Acid Exposure
Citation:
Saili, K., T. Zurlinden, T. Antonijevic, I. Shah, C. Deisenroth, AND T. Knudsen. Modeling Endodermal Differentiation Trajectories Following All-trans Retinoic Acid Exposure. Presented at FutureToxIV, Crystal City, VA, November 14 - 16, 2018. https://doi.org/10.23645/epacomptox.7752452
Impact/Purpose:
In silico modeling of this phenotypic shift will serve as a basis for testing additional chemicals for potential disruption to early embryonic development.
Description:
Numerous organ systems originate from the embryonic endoderm, including the thyroid, thymus, lungs, stomach, liver, pancreas, small and large intestines. We monitored differentiating endoderm for 8 days following all-trans retinoic acid (ATRA) exposure to identify early molecular events that may lead to malformations following exposure to a teratogen. Induced pluripotent stem cells derived from adult female fibroblasts were directed to a differentiating trajectory using low serum and activin. Cells were then exposed daily to 5 concentrations of ATRA (0.001 to 10 µM) or 0.1% DMSO control and collected at three timepoints (6h, 96h, and 192h) for high content imaging (HCI) and RNA-sequencing analysis. Cell count and FOXA2 expression as a marker of definitive endoderm were measured by HCI. A tipping point defined by the persistence of effects at both 96h and 192h timepoints was identified at 0.1µM. Concentrations above 0.1µM caused an increase in cell counts and reduced FOXA2 expression compared to time-matched controls. Global gene expression at concentrations above and below the tipping point followed two diverging trajectories. Developmental marker expression suggested that cells exposed to concentrations above 0.1µM transiently reverted to pluripotent states followed by a shift in endodermal fate from an anterior to posterior molecular phenotype. In silico modeling of this phenotypic shift will serve as a basis for testing additional chemicals for potential disruption to early embryonic development. This work does not represent US EPA policy.
URLs/Downloads:
DOI: Modeling Endodermal Differentiation Trajectories Following All-trans Retinoic Acid Exposure
SAILI_FUTURETOXIV_POSTER_181109_M.PDF (PDF, NA pp, 1361.168 KB, about PDF)