OP11 Organoids derived from inflamed intestinal biopsies of patients with ulcerative colitis lose their inflammatory phenotype during
ex vivo culture
K. Arnauts*1,2, B. Verstockt1,3, M. Vancamelbeke1, S. Vermeire1,3, C. Verfaillie2, M. Ferrante1,3
1KU Leuven, Department of Chronic Diseases, Metabolism and Ageing (CHROMETA), Leuven, Belgium, 2KU Leuven, Department of Development and Regeneration, Leuven, Belgium, 3KU Leuven, Department of Gastroenterology and Hepatology, Leuven, Belgium
Patient-derived intestinal organoids provide an excellent tool to unravel the multi-factorial mechanisms underlying ulcerative colitis (UC). Organoids develop from stem cell-containing intestinal crypts and recapitulate many features of the source tissue. However, it remains unclear whether
Fresh biopsies from both inflamed and non-inflamed segments were obtained during endoscopy from eight patients with active UC (endoscopic Mayo sub-score of ≥2) and an accessible border of inflammation. Crypts were isolated and cultured as organoids for 4 weeks with weekly mechanical splitting. RNA was extracted from biopsies, crypts, and 1- and 4-week-old organoids. RNA sequencing was performed by Lexogen QuantSeq for Illumina. Differential gene expression and pathways were studied through DESeq2 and Ingenuity Pathway Analysis (FDR < 0.05).
Biopsies and crypts from inflamed regions showed separate clustering on principal component analysis (PCA, Figure) and significantly higher activation of inflammatory pathways, including antigen presentation (
We conclude that
Principal component analysis (PCA) shows separate clustering of biopsies and crypts from inflamed regions vs. non-inflamed regions. After 1 and 4 weeks, organoids of inflamed and non-inflamed origin cluster together and are no longer distinguishable.