PREDICTING HUMAN DEVELOPMENTAL TOXICITY OF PHARMACEUTICALS USING HUMAN STEM-LIKE CELLS AND METABOLOMICS

摘要

The invention provides biomarker profiles of metabolites and methods for screening chemical compounds including pharmaceutical agents, lead and candidate drug compounds and other chemicals using human stem-like cells (hSLCs) or lineage-specific cells produced therefrom. The inventive methods are useful for testing toxicity, particularly developmental toxicity and detecting teratogenic effects of such chemical compounds. Specifically, a more predictive developmental toxicity model, based on an in vitro method that utilizes both hSLCs and metabolomics to discover biomarkers of developmental toxicity is disclosed.

主权项

1. A method of predicting teratogenicity of a test compound, comprising the steps of:
(a) culturing hSLCs:
(i) in the presence of a first known teratogenic compound; and(ii) in the absence of the first known teratogenic compound; (b) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the first known teratogenic compound in comparison with hSLCs not exposed to the first known teratogenic compound in order to identify a difference in metabolic response of hSLCs exposed to the first known teratogenic compound in comparison with hSLCs not exposed to the first known teratogenic compound; (c) analyzing the difference in metabolic response in order to generate a set of mass features associated with exposure of hSLCs to the first teratogenic compound; (d) repeating steps a)-c) multiple times, each time with a different known teratogenic compound; (e) grouping mass features generated from each exposure to a teratogenic compound to obtain a first reference profile of mass features; (f) comparing a profile of mass features generated upon exposure of hSLCs to a test compound with the first reference profile to predict the teratogenicity of the test compound; (g) if the test compound is predicted to be a teratogen, adding the profile of mass features to the first reference profile to obtain a second reference profile, wherein the predictive accuracy of the second reference profile is greater than the predictive accuracy of the first reference profile; and (h) repeating steps f) and g) multiple times, each time with a different test compound to obtain a final reference profile.