Evaluation of developmental toxicants and signaling pathways in a functional test based on the migration of human...

Area of Science:

• Toxicology
• Developmental Biology
• Stem Cell Biology

Background:

• Limited data exists on the developmental toxicity (DT) of many chemicals, necessitating high-throughput testing alternatives.
• Animal testing for DT is resource-intensive, driving the need for novel in vitro screening strategies.
• Impaired neural crest (NC) cell function is a known mechanism underlying teratogenic effects, highlighting the importance of NC-based assays for DT assessment.

Purpose of the Study:

• To develop a human-relevant neural crest (NC) cell function assay for sensitive screening of environmental toxicants.
• To establish a robust assay capable of identifying toxicity pathways involved in NC cell dysfunction.

Main Methods:

• Human embryonic stem cells were differentiated into NC cells.
• A migration assay for NC cells (MINC assay) was established and validated.
• Environmental toxicants and pathway inhibitors were tested using the MINC assay, coupled with gene expression profiling.

Main Results:

• The MINC assay demonstrated high sensitivity to known NC toxicants like methylmercury, valproic acid, and lead acetate.
• The assay accurately identified specific NC toxicants (triadimefon, triadimenol) and differentiated sensitivities to mercury compounds.
• Analysis revealed key signaling pathways critical for NC cell migration, elucidated through pharmacologic inhibition and gene expression analysis.

Conclusions:

• The MINC assay serves as a reliable model for human NC cell migration.
• The assay exhibits high sensitivity and specificity in detecting developmental toxicant-induced impairment of NC cell function.
• This assay provides a valuable tool for screening environmental toxicants and understanding their impact on NC cell-mediated developmental processes.