DOP04 The ability of epithelial regeneration is reduced in the Crohn's disease patient-derived organoids, especially in TNF-α enriched condition. (25th January 2019)

• Record Type: Journal Article
• Authors: Song, J H; Hong, S N; Seong, G; Kim, T J; Kim, E R; Chang, D K; Kim, Y-H; Lee, C; Lee, C
• Is Part Of: Journal of Crohn's and colitis. Volume 13(2019)Supplement 1
• Page Start: S029
• Page End: S030
• Publication Date: 2019-01-25
• Subjects: Inflammatory bowel diseases -- Periodicals; 616.344005
• DOI: 10.1093/ecco-jcc/jjy222.039
• Languages: English
• ISSNs: 1873-9946

Abstract

Abstract: Background: Recent evidence has featured 'mucosal healing' as the most significant prognostic factor for long-term remission in patients with Crohn's disease (CD), suggesting that accomplishment of epithelial regeneration is critically required to improve the treatment for CD. Recent established minigut organoid model re-enact the epithelial layer of patients and provides and is able to assess the ability of epithelial regeneration. Methods: Intestinal crypts were isolated from jejunum and ileum using the endoscopic biopsy of normal mucosa of CD patients and healthy controls and performed the ex vivo 3-dimensional culture. One hundred crypts were plated on Matrigel and counted the organoid formation (CD: duodenum, n = 15, jejunum, n = 18, ileum, n = 21, colon, n = 5, controls: duodenum, n = 16, jejunum, n = 16, ileum, n = 12, colon, n = 12). The organoids maintained in culture for at least 8 passages (stable organoids, CD: jejunum, n = 5, ileum, n = 3, controls: jejunum, n = 8, ileum, n = 5). Stable enteroids derived from CD patients and controls were cultured with different concentration of TNF-α and assessed the organoid morphology. After Day 7 days of culture, viability via MTT reduction, proliferation via Edu incorporation, apoptosis via TUNEL stain, and histology of patient-derived organoids were compared with those of controls. Results: The organoid formation rate of crypt of CD patients were significantly reduced those of controls (duodenum: 53 ± 18 vs. 62 ± 12, p = 0.652; jejunum: 38 ± 15% vs. 58 ± 16%, p < 0.05; ileum: 48 ± 15 vs. 21 ± 8%, p < 0.01; and colon: 24 ± 10 vs. 12 ± 8%, p < 0.01). When the CD patient-derived organoids maintained more than 4–6 passage, the formation rate and morphology of organoid were parallel in those of controls. Stable organoids passaged at least 8 times were counted before TNF-α treatment at the budding stage after 3 days of culture. With additional 7 days culture with TNF-α (0 ng/ml, 10 ng/ml, 30 ng/ml, and 100 mg/ml), the relative number of organoids and enteroid/spheroid ratio were decreased steadily as increasing concentration of TNF-α. Susceptibility of TNF-α was significantly amplified in CD patients-derived organoids compared with normal organoids and ileal organoids compared with jejunal organoids. MTT-stained organoids were decreased and Edu-positive cells and TUNEL-positive cells were increased in CD ileal organoids significantly. Conclusions: The ability of epithelial regeneration is reduced in the CD patients, especially in TNF-α enriched condition.