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
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
We aimed to establish an
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.
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.
The establishment of
1. Hibiya S, Tsuchiya K, Hayashi R,