Establishment of a system to evaluate the therapeutic effect and the dynamics of an investigational drug on ulcerative colitis using human colonic organoids
Ulcerative colitis (UC) is a chronic inflammatory disease of the colon with an intractable, recurrent course. The goal of UC therapy is to target mucosal healing because immune-suppressive therapy for UC frequently results in relapse. However, few drugs directly target mucosal healing. We, therefore, aim to evaluate the therapeutic effect of an investigational drug on intestinal epithelial cells in an in vitro UC model using human colonic organoids.
Colonic organoids were isolated from human colon and cultured. A mixture of cytokines and bacterial components were used to mimic UC in humans. The effect of the investigational drug on colonic organoid was evaluated by microarray analysis and 3D immunofluorescence. The enrichment of stem cells was assessed with a colony formation assay.
Inflammatory stimulation resulted in a significant induction of inflammatory-related genes in colonic organoids whereas cell differentiation was suppressed. Treatment with the investigational drug KAG-308 showed reciprocal dynamics of gene expression to inflammatory stimulation, which resulted in not only the suppression of immune response but also the promotion of cellular differentiation towards secretory lineages. Moreover, SPDEF and Reg4 were identified as novel targets for the enrichment of intestinal epithelial stem cells and mucosal healing.
The establishment of in vitro UC model using human colonic organoid could reveal the effects and targets of investigational drugs in intestinal epithelial cells under inflammation conditions. Further maturation of this system might be more efficient to predict the effect on UC, as compared with the use of animal model, for the development of new drugs.
KeywordsInflammatory bowel disease In vitro human model Evaluation system Investigational drug
Inflammatory bowel disease
Dextran sulphate sodium
Intestinal epithelial cells
Prostaglandin E2 receptor 4
Gene set enrichment analysis
Reactive oxygen species
Tumour necrosis factor
Class II major histocompatibility complex transactivator
Regenerating family member 4
SAM-pointed domain containing ETS transcription factor
Dual oxidase maturation factor 2
Aldehyde dehydrogenase 1
Leucine-rich repeat-containing G-protein coupled receptor 5
Hairy and enhancer of split 1
Atonal bHLH transcription factor 1
Trefoil factor 3
This work was supported by scientific Research, from the Japanese Ministry of Education, Culture, Sports, Science and Technology as follows (KAKENHI grant numbers 24590935, 25114703, 25130704, 26221307, 15H04808, 16H06770, 17H06654, 17K15930, 17K19513); Japan Foundation for Applied Enzymology; the Health and Labor Sciences Research Grant from the Japanese Ministry of Health, Labor and Welfare (MHLW) as follows (Grant number 14526073); The Practical Research for Innovative Cancer Control, Rare/Intractable Diseases from Japan Agency for Medical Research and Development (AMED) as follows (Grant numbers 15Ack0106017h0002, 16ck0106017h0003, 16ek0109456h0003); Research Grant of the Princess Takamatsu Cancer Research Fund; Naoki Tsuchida Research Grant.
Compliance with ethical standards
Conflict of interest
YM and YW are employed by Kaken Pharmaceutical Co., Ltd. KT has received research funding (collaborative research) provided by Kaken Pharmaceutical Co., Ltd.
MW, TN and KT designed the experiments and acquired data. TS and SH established inflammatory stimulation of the organoids. RN and SW assessed the effect of KAG-308 on the organoids. YM and YW assessed the effect of KAG-308 on DSS colitis model in mice. KT and MW coordinated the projects and drafted the manuscript.
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