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Updated: May 22, 2025
Author Spotlight: Investigating the Effects of Compounds on Intestinal Tissue Using 3D Human Cell Line Models
Published on: September 1, 2023
High-throughput assay for predicting diarrhea risk using a 2D human intestinal stem cell-derived model
Colleen M Pike1, James A Levi1, Lauren A Boone1
1Altis Biosystems, Durham, NC, 27709, USA.
Abstract:
Gastrointestinal toxicities (GITs) in clinical trials often lead to dose-limitations that reduce drug efficacy and delay treatment optimization. Preclinical animal models do not accurately replicate human physiology, leaving few options for early detection of GITs, such as diarrhea, before human studies. Chemotherapeutic agents, known to cause clinical diarrhea, frequently target mitotic cells. Therefore, we hypothesized a model utilizing proliferative cell populations derived from human intestinal crypts would predict clinical diarrhea occurrence with high accuracy. Here, we describe the development of a diarrhea prediction assay utilizing RepliGut® Planar, a primary intestinal stem cell-derived platform. To evaluate the ability of this model to predict clinical diarrhea risk, we assessed toxicity of 30 marketed drugs by measuring cell proliferation (EdU incorporation), cell abundance (nuclei quantification), and barrier formation (TEER) in cells derived from three human donors. Dose response curves were generated for each drug, and the IC15 to Cmax ratio was used to identify a threshold for assay positivity. This model accurately predicted diarrhea potential, achieving an accuracy of 91 % for proliferation, 90 % for abundance, and 88 % for barrier formation. In vitro toxicity screening using primary proliferative cells may reduce clinical diarrhea and ultimately lead to safer and more effective treatments for patients.
Insights
A new human intestinal stem cell model accurately predicts drug-induced diarrhea. This in vitro assay uses RepliGut® Planar to assess toxicity, improving drug safety and efficacy for patients.
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.
