Abstract
Exposure to teratogenic chemicals during pregnancy may cause severe birth defects. Due to high inter-species variation of drug responses as well as financial and ethical burdens, despite the widely use of in vivo animal tests, it’s crucial to develop highly predictive human pluripotent stem cell (hPSC)-based in vitro assays to identify potential teratogens. Previously we have shown that the morphological disruption of mesoendoderm patterns formed by geometrically-confined cell differentiation and migration using hPSCs could potentially serve as a sensitive morphological marker in teratogen detection. Here, a micropatterned human pluripotent stem cell test (µP-hPST) assay was developed using 30 pharmaceutical compounds. A simplified morphometric readout was developed to quantify the mesoendoderm pattern changes and a two-step classification rule was generated to identify teratogens. The optimized µP-hPST could classify the 30 compounds with 97% accuracy, 100% specificity and 93% sensitivity. Compared with metabolic biomarker-based hPSC assay by Stemina, the µP-hPST could successfully identify misclassified drugs Bosentan, Diphenylhydantoin and Lovastatin, and show a higher accuracy and sensitivity. This scalable µP-hPST may serve as either an independent assay or a complement assay for existing assays to reduce animal use, accelerate early discovery-phase drug screening and help general chemical screening of human teratogens.
Highlights
Exposure to teratogenic chemicals during pregnancy may cause severe birth defects
We developed a two-step classification rule for the teratogen identification to establish the in vivo relevance of the compound screening in our micropatterned human pluripotent stem cell test
Similarity in cytotoxicity profile could be found in compounds belonging to the same class of chemical property or therapeutic use, especially in terms of general cytotoxicity
Summary
Exposure to teratogenic chemicals during pregnancy may cause severe birth defects. Due to high interspecies variation of drug responses as well as financial and ethical burdens, despite the widely use of in vivo animal tests, it’s crucial to develop highly predictive human pluripotent stem cell (hPSC)-based in vitro assays to identify potential teratogens. We developed a two-step classification rule for the teratogen identification to establish the in vivo relevance of the compound screening in our micropatterned human pluripotent stem cell test (μP-hPST) This refined and validated assay with high predictability for teratogen detection would find wide applications in drug development and as a cost-saving assay for early detection of teratogens in industrial chemicals, household and consumer goods, food and nutraceuticals, cosmetics and environmental toxins. This human cell-based in vitro assay will resolve the inter-species variation problems of the existing assays and contribute greatly to the reduction of animal uses
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