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P026 Establishment of an in vitro system to evaluate the therapeutic effect of the investigational drug on ulcerative colitis using human colonic organoids

S. Hibiya*1, K. Tsuchiya1, R. Nishimura1, T. Shirasaki1, S. Watanabe1, N. Katsukura1, T. Nakamura2, M. Watanabe1

1Tokyo Medical and Dental University, Gastroenterology and Hepatology, Tokyo, Japan, 2Tokyo Medical and Dental University, Advanced Therapeutics for Gastrointestinal Diseases, Tokyo, Japan

Background

The goal of ulcerative colitis (UC) therapy has recently been to target mucosal healing. However, few drugs to directly target mucosal healing have been developed. Although the effects of investigational drugs can be estimated using in vivo mouse colitis model, it is unclear whether the target of the drugs is inflammation or mucosal damage. The establishment of an in vitro system to evaluate mucosal regeneration is necessary for targeting mucosal healing. We have established an in vitro chronic inflammation model by using mice colonic organoids.1

We aimed to establish an in vitro human model for chronic colitis using human primary colon organoids and to evaluate therapeutic effect of an investigational drug.

Methods

This study was approved by the Ethics Committee. Human colonic organoids were generated from non-inflamed colon. The mixture of inflammatory reagents was added into the medium to mimic UC. Gene Set Enrichment Analysis (GSEA) was performed for the comparison between organoids and biopsies from active UC patients. To assess the effect of investigational drug (KAG-308) on intestinal epithelial cells under chronic inflammation, the drug was added into the medium for 1 week. The effect of the drug was evaluated by microarray analysis, colony formation assay and proliferation assay. The molecular target of the drug was identified by microarray analysis. Colitis was induced in mice by 1.5% dextran sulphate sodium (DSS) in drinking water for 6 days and subsequently providing 0.5% DSS for 4 days.

Results

Microarray analysis of the inflamed organoids showed significant induction of inflammatory signalling-related genes. GSEA showed the similarities of up-regulated genes in between inflamed human organoids and biopsies from active UC patients, suggesting that the inflamed organoids might acquire UC like phenotype. Treatment with an investigational drug showed reciprocal dynamics of gene expression to inflammatory stimulation, suggesting this drug has antagonistic functions against chronic inflammation. Moreover, the genes which have reciprocal dynamics of its expression between inflammatory stimulation and drug treatment were identified as molecular target of this drug. Treatment with this drug also promoted stem cell population and cell proliferation of the organoids even under the inflammatory situation. Finally, this drug-induced mucosal healing in subacute DSS colitis model mice.

Conclusion

The establishment of in vitro chronic colitis model using human colonic organoid could reveal the effects and targets of an investigational drug in intestinal epithelial cells. Further maturation of this system might be more efficient to predict the effect on UC for the development of new drugs.

Reference

1. Hibiya S, Tsuchiya K, Hayashi R, et al. Long-term inflammation transforms intestinal epithelial cells of colonic organoids. J Crohns Colitis 2017;5:621–30. doi:10.1093/ecco-jcc/jjw186