Drug-induced gastrointestinal toxicity and barrier integrity: cytoskeleton-mediated impairment in a clinically...

Area of Science:

• Gastroenterology
• Drug Development
• Cell Biology

Background:

• Drug-induced gastrointestinal (GI) toxicity is a significant challenge in drug development, often poorly predicted by current assays.
• Conventional Caco-2 cell assays lack the physiological relevance needed for accurate toxicity prediction.

Purpose of the Study:

• To evaluate a novel transepithelial electrical resistance (TEER) assay using human intestinal epithelial cells (hIECs) for predicting drug-induced GI toxicity.
• To compare the predictive performance of the hIEC TEER assay against traditional cell viability and Caco-2 TEER assays.

Main Methods:

• Developed and utilized a TEER assay with nontransformed hIECs derived from human pluripotent stem cells.
• Tested 17 clinically relevant compounds, including cell cycle inhibitors, tyrosine kinase inhibitors, and NSAIDs.
• Quantified drug exposure using margin of safety calculations and performed transcriptomic analysis for mechanistic insights.

Main Results:

• The hIEC TEER assay demonstrated superior predictive accuracy (AUC 0.96) compared to Caco-2 TEER (AUC 0.72) and cell viability assays (AUC ≤0.69).
• Achieved high sensitivity (92%), specificity (100%), and accuracy (94%) in predicting GI toxicity.
• Transcriptomic analysis revealed that microtubule-targeting chemotherapy downregulates cytoskeleton-related pathways, impairing intestinal barrier function.

Conclusions:

• The hIEC TEER assay is a robust platform for accurately predicting drug-induced GI toxicity.
• This assay provides mechanistic insights into cytoskeleton-mediated barrier impairment, facilitating early identification of GI toxicity during drug development.