A high throughput screening system for predicting chemically-induced reproductive organ deformities

Area of Science:

• Toxicology
• Biochemistry
• Genetics

Background:

• Assessing reproductive toxicants requires practical, cost-effective, and rapid alternative testing methods.
• Previous pragmatic approaches, while successful, were complex and low-throughput.
• There is a need for simplified, high-throughput screening batteries for reproductive toxicity.

Purpose of the Study:

• To simplify a reproductive toxicant screening battery using a mechanistic approach.
• To validate a high-throughput battery of CALUX (Chemical-Activated Luciferase eXpression) reporter gene assays.
• To assess the battery's ability to predict specific classes of reproductive toxicity.

Main Methods:

• Utilized a mechanistic approach to validate the screening battery.
• Employed a panel of high-throughput CALUX reporter gene assays.
• Challenged the battery with preselected chemical sets targeting reproductive organ deformities and neural tube defects.

Main Results:

• The simplified battery demonstrated selectivity in predicting toxicity.
• Correctly predicted 82% of compounds inducing reproductive organ deformities.
• Correctly predicted 47% of compounds inducing neural tube defects.
• Identified ERalpha CALUX (estrogen receptor alpha) and AR-anti CALUX (androgen receptor antagonism) assays as most informative.

Conclusions:

• The developed high-throughput CALUX assay battery offers a simplified and mechanistic approach to screening reproductive toxicants.
• The battery shows promise in predicting specific reproductive toxicity endpoints, particularly organ deformities.
• Estrogenicity and androgen receptor antagonism assays are crucial components for assessing reproductive toxicity.