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Updated: Dec 30, 2025
Human Pluripotent Stem Cell Based Developmental Toxicity Assays for Chemical Safety Screening and Systems Biology Data Generation
Published on: June 17, 2015
Screening ToxCast™ for Chemicals That Affect Cholesterol Biosynthesis: Studies in Cell Culture and Human Induced
Phillip A Wages1, Piyush Joshi2, Keri A Tallman1
1Department of Chemistry and Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, Tennessee, USA.
Background:
Changes in cholesterol metabolism are common hallmarks of neurodevelopmental pathologies. A diverse array of genetic disorders of cholesterol metabolism support this claim as do multiple lines of research that demonstrate chemical inhibition of cholesterol biosynthesis compromises neurodevelopment. Recent work has revealed that a number of commonly used pharmaceuticals induce changes in cholesterol metabolism that are similar to changes induced by genetic disorders with devastating neurodevelopmental deficiencies.
Objectives:
We tested the hypothesis that common environmental toxicants may also impair cholesterol metabolism and thereby possibly contribute to neurodevelopmental toxicity.
Methods:
Using high-throughput screening with a targeted lipidomic analysis and the mouse neuroblastoma cell line, Neuro-2a, the ToxCast™ chemical library was screened for compounds that impact sterol metabolism. Validation of chemical effects was conducted by assessing cholesterol biosynthesis in human induced pluripotent stem cell (hiPSC)-derived neuroprogenitors using an isotopically labeled cholesterol precursor and by monitoring product formation with UPLC-MS/MS.
Results:
Twenty-nine compounds were identified as validated lead-hits, and four were prioritized for further study (endosulfan sulfate, tributyltin chloride, fenpropimorph, and spiroxamine). All four compounds were validated to cause hypocholesterolemia in Neuro-2a cells. The morpholine-like fungicides, fenpropimorph and spiroxamine, mirrored their Neuro-2a activity in four immortalized human cell lines and in a human neuroprogenitor model derived from hiPSCs, but endosulfan sulfate and tributyltin chloride did not.
Conclusions:
These data reveal the existence of environmental compounds that interrupt cholesterol biosynthesis and that methodologically hiPSC neuroprogenitor cells provide a particularly sensitive system to monitor the effect of small molecules on de novo cholesterol formation. https://doi.org/10.1289/EHP5053.
Insights
Environmental toxicants can disrupt cholesterol metabolism, potentially leading to neurodevelopmental toxicity. This study identified specific compounds that impair cholesterol biosynthesis, highlighting sensitive human cell models for future research.
Area of Science:
- Environmental toxicology
- Neuroscience
- Biochemistry
Background:
- Cholesterol metabolism disruptions are linked to neurodevelopmental disorders.
- Genetic and pharmaceutical factors affecting cholesterol biosynthesis impact neurodevelopment.
- Environmental toxicants are investigated for their potential role in neurodevelopmental toxicity via cholesterol pathways.
Purpose of the Study:
- To test if common environmental toxicants impair cholesterol metabolism.
- To identify environmental compounds that affect sterol metabolism and could contribute to neurodevelopmental toxicity.
Main Methods:
- High-throughput screening of the ToxCast™ chemical library using Neuro-2a cells and targeted lipidomic analysis.
- Validation of identified compounds using human induced pluripotent stem cell (hiPSC)-derived neuroprogenitors to assess cholesterol biosynthesis.
- Utilized UPLC-MS/MS for monitoring cholesterol precursor product formation.
Main Results:
- Twenty-nine validated lead-hit compounds impacting sterol metabolism were identified.
- Four compounds (endosulfan sulfate, tributyltin chloride, fenpropimorph, spiroxamine) were prioritized, all causing hypocholesterolemia in Neuro-2a cells.
- Fenpropimorph and spiroxamine showed consistent effects in human cell lines and hiPSC-derived neuroprogenitors, unlike endosulfan sulfate and tributyltin chloride.
Conclusions:
- Environmental compounds capable of interrupting cholesterol biosynthesis have been identified.
- Human induced pluripotent stem cell (hiPSC)-derived neuroprogenitors are a sensitive model for monitoring the effects of small molecules on cholesterol formation.
- This research links environmental toxicants to cholesterol metabolism disruption and potential neurodevelopmental impacts.
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