Search

P046 Disease modelling of Inflammatory Bowel Disease by human colon organoids

Molnár, T.(1,2);Jójárt, B.(1,2,3);Resál, T.(1);Szántó, K.(1);Kata, D.(4);Földesi, I.(4);Molnár, T.(1);Maléth, J.(1,2,3,5);Farkas, K.(1);

(1)University of Szeged, First Department of Medicine, Szeged, Hungary;(2)Hungarian Academy of Science, University of Szeged Momentum Epithelial Cell Signaling and Secretion Research Group, Szeged, Hungary;(3)Hungarian Centre of Excellence of Molecular Medicine, University of Szeged Molecular Gastroenterology Research Group, Szeged, Hungary;(4)University of Szeged, Institute of Laboratory Medicine, Szeged, Hungary;(5)University of Szeged, Department of Public Health, Szeged, Hungary

Background

Inflammatory bowel diseases (IBD) are characterized by chronic inflammation of the gastrointestinal tract which is associated with the imbalanced pro- and anti-inflammatory cytokines. Anti-TNF-α is widely used as a therapeutic agent, but 10-30% of the patients are not responding to the treatment or in time it become less affective. In vitro organoid cultures generated from stem cells can mimic the cellular diversity and function of the organ of origin and might be used as a predictive tool for patient specific response. However, the in vitro cytokine expression pattern of the IBD colon organoid cultures have never been determined or compared with the cytokine expression of the colon mucosa.

Methods

Our aim was to determine and compare the cytokine profiles of the colon biopsy samples in IBD patients and organoid cultures generated from these biopsies.
In this study samples from 5 IBD patients were used. Biopsies were taken during colonoscopy from inflamed part of the colon. Crypts were isolated from the biopsy samples to start colon organoid culture. Total protein was isolated from the biopsies and the organoids. Cytokine Array was used to determine the cytokine patterns in both samples. ELISA was used to measure the TNF-α concentrations and immunofluorescence staining to localize it in the organoids.

Results

We determined the cytokine profile of the biopsy samples and the colon organoid cultures. The cytokine patterns were remarkably similar until the first passage. The major pro-inflammatory cytokines, IL-1β, IL-6, IL-8, were detected in the organoids after the first passage. After the second passage the cytokine expression decreased or disappeared. ELISA revealed that TNF-α was also expressed in the organoid cultures, however a similar decrease in the expression was observed, which was also confirmed by immunofluorescence staining. TNF-α positive cells were present in the organoids, but it decreased after every passage.

Conclusion

Our results suggest that the colon organoid cultures maintain the cytokine profile of the tissue of origin until a limited time (until the first passage). Therefore, organoids might be used to determine patient specific therapy for drug tests.

Portal