Predicting human developmental toxicity of pharmaceuticals using human stem-like cells and metabolomics
First Claim
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1. A method of predicting human developmental toxicity of a test compound, the method comprising the steps of:
- (a) culturing human stem-cell like cells (hSLCs);
(i) in the presence of a first known developmental toxicant; and
(ii) in the absence of the first known developmental toxicant;
(b) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant in order to identify a difference in metabolic response of hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant;
wherein the plurality of metabolites is detected by mass spectrometry;
(c) generating a set of mass features associated with exposure of hSLCs to the first developmental toxicant by analyzing the difference in metabolic response;
(d) repeating steps a)-c) multiple times, each time with a different known developmental toxicant;
(e) grouping mass features generated from each exposure to a developmental toxicant to obtain a reference profile of mass features correlating with developmental toxicity;
(f) culturing hSLCs;
(i) in the presence of a test compound; and
(ii) in the absence of the test compound;
(g) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the test compound in comparison with hSLCs not exposed to the test compound in order to identify a difference in metabolic response of hSLCs exposed to the test compound in comparison with hSLCs not exposed to the test compound;
wherein the plurality of metabolites is detected by mass spectrometry; and
(h) generating a set of mass features associated with exposure of hSLCs to the test compound by analyzing the difference in metabolic response;
(i) comparing the set of mass features associated with exposure of hSLCs to the test compound of step (h) to the reference profile of mass features correlating with developmental toxicity of step (e) to predict the human developmental toxicity of the test compound.
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Abstract
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.
17 Citations
22 Claims
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1. A method of predicting human developmental toxicity of a test compound, the method comprising the steps of:
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(a) culturing human stem-cell like cells (hSLCs); (i) in the presence of a first known developmental toxicant; and (ii) in the absence of the first known developmental toxicant; (b) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant in order to identify a difference in metabolic response of hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant; wherein the plurality of metabolites is detected by mass spectrometry; (c) generating a set of mass features associated with exposure of hSLCs to the first developmental toxicant by analyzing the difference in metabolic response; (d) repeating steps a)-c) multiple times, each time with a different known developmental toxicant; (e) grouping mass features generated from each exposure to a developmental toxicant to obtain a reference profile of mass features correlating with developmental toxicity; (f) culturing hSLCs; (i) in the presence of a test compound; and (ii) in the absence of the test compound; (g) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the test compound in comparison with hSLCs not exposed to the test compound in order to identify a difference in metabolic response of hSLCs exposed to the test compound in comparison with hSLCs not exposed to the test compound; wherein the plurality of metabolites is detected by mass spectrometry; and (h) generating a set of mass features associated with exposure of hSLCs to the test compound by analyzing the difference in metabolic response; (i) comparing the set of mass features associated with exposure of hSLCs to the test compound of step (h) to the reference profile of mass features correlating with developmental toxicity of step (e) to predict the human developmental toxicity of the test compound. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20)
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21. A method of producing a reference profile of mass features correlating with developmental toxicity, the method comprising the steps of:
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(a) culturing human stem-cell like cells (hSLCs); (i) in the presence of a first known developmental toxicant; and (ii) in the absence of the first known developmental toxicant; (b) detecting a plurality of metabolites having a molecular weight of less than about 3000 Daltons associated with hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant in order to identify a difference in metabolic response of hSLCs exposed to the first known developmental toxicant in comparison with hSLCs not exposed to the first known developmental toxicant; wherein the plurality of metabolites is detected by mass spectrometry; (c) generating a set of mass features associated with exposure of hSLCs to the first developmental toxicant by analyzing the difference in metabolic response; (d) repeating steps a)-c) multiple times, each time with a different known developmental toxicant; (e) grouping mass features generated from each exposure to a developmental toxicant to obtain a reference profile of mass features correlating with developmental toxicity. - View Dependent Claims (22)
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Specification