High-Throughput Assay for Predicting Diarrhea Risk Using a 2D Human Intestinal Stem Cell-Derived Model.

Area of Science:

• Drug discovery and development
• Toxicology
• Gastroenterology

Background:

• Gastrointestinal toxicities (GITs) are common adverse events (AEs) in clinical trials, often limiting drug efficacy and development.
• Current preclinical models fail to accurately predict human GITs due to differences in physiology.
• Early and accurate prediction of GITs is crucial for successful clinical trial outcomes and optimized patient treatments.

Purpose of the Study:

• To develop and validate a novel in vitro model for predicting clinical gastrointestinal toxicities.
• To assess the utility of a multiparametric assay using human intestinal stem cell-derived cultures for early GIT detection.
• To evaluate the model's accuracy in predicting diarrhea risk for marketed chemotherapeutics.

Main Methods:

• Development of a multiparametric assay using the RepliGut® Planar system in a Transwell™ format.
• Assessment of cell proliferation (EdU incorporation), cell abundance (DAPI quantification), and barrier function (TEER) in response to drug treatment.
• Evaluation of 30 drugs with known clinical diarrhea incidence across three human donors.

Main Results:

• The developed assay demonstrated reproducible responses of primary proliferative cells to chemotherapeutics at physiologic concentrations.
• Highly accurate predictions of diarrhea potential were achieved for all endpoints: 91% (DAPI), 90% (EdU), and 88% (TEER).
• Minimal variation was observed across different human donors, indicating model robustness.

Conclusions:

• In vitro toxicity screening using primary proliferative cells offers a promising approach for improved drug safety evaluations.
• This model can potentially reduce adverse events in clinical trials, leading to safer and more effective patient treatments.
• Early detection of GITs through this advanced in vitro system supports optimized drug development pathways.