High-throughput assay for predicting diarrhea risk using a 2D human intestinal stem cell-derived model.

Area of Science:

• Gastroenterology
• Toxicology
• Drug Development

Background:

• Gastrointestinal toxicities (GITs) limit drug efficacy and delay treatment optimization in clinical trials.
• Preclinical animal models fail to accurately replicate human physiology, hindering early detection of GITs like diarrhea.
• Chemotherapeutic agents often target mitotic cells, a mechanism linked to clinical diarrhea.

Purpose of the Study:

• To develop and validate a predictive assay for clinical diarrhea using human intestinal stem cell-derived cultures.
• To assess the accuracy of a novel in vitro model in predicting diarrhea risk associated with marketed drugs.

Main Methods:

• Development of a diarrhea prediction assay using RepliGut® Planar, a platform derived from primary human intestinal stem cells.
• Evaluation of 30 marketed drugs by measuring cell proliferation (EdU incorporation), cell abundance (nuclei quantification), and barrier formation (TEER).
• Generation of dose-response curves and calculation of the IC15 to Cmax ratio to determine assay positivity.

Main Results:

• The model demonstrated high accuracy in predicting diarrhea potential: 91% for proliferation, 90% for abundance, and 88% for barrier formation.
• Dose-response curves and IC15 to Cmax ratios were established for each drug, defining a threshold for assay positivity.
• The assay successfully identified diarrhea risk across a range of marketed drugs.

Conclusions:

• In vitro toxicity screening using primary human intestinal stem cell-derived models can accurately predict clinical diarrhea.
• This approach may reduce the incidence of clinical diarrhea, leading to safer and more effective drug treatments.
• The RepliGut® Planar platform offers a promising tool for early detection of drug-induced gastrointestinal toxicities.