Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Pammer, A.
The Endocannabinoid System in Eosinophilic Esophagitis.
[ Diplomarbeit/Master Thesis (UNI) ] TU Graz; 2022. pp.55.
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- Autor*innen der Med Uni Graz:
- Betreuer*innen:
- Gruden Eva
- Schicho Rudolf
- Altmetrics:
- Abstract:
- Eosinophilic esophagitis (EoE) is a chronic antigen-driven inflammatory disease characterized by increased interleukin 13 (IL-13) expression. Patients suffer from symptoms such as difficulty in swallowing, food impaction, regurgitations as well as vomiting and chest pain leading to drastical restrictions in their daily life. Studies have shown, that forkhead box P3 (FoxP3) as well as cannabinoid receptor 2 (CB2) expression are increased while monoacylglycerol lipase (MGL) expression is decreased in mucosal esophageal biopsies of active EoE. Therefore, we set out to investigate the involvement of the endocannabinoid system in EoE pathology. The study consisted of investigating three different types of cells. Firstly, CD4+ T cells were inspected with regard to the contribution of CB2 receptor in regulatory T cell (Treg) differentiation as well as the impact of CB2 antagonism and IL-13 to regulatory T cell suppression capacity was explored. Moreover, structural cells that contribute to inflammation and disease progression were studied and tested for MGL expression. As part of our first aim, CD4+ T cells were differentiated into Tregs while blocking the CB2 receptor or activating it, which was followed by measuring the Treg (CD4+FoxP3+) percentage with flow cytometry (FACS). Supernatants were tested for transforming growth factor - using ELISA technology and the suppression ability of Tregs was investigated by measuring the proliferation of prestained CD4+ T cells via flow cytometry. The results indicate that CB2 activation contributes to Treg differentiation via transforming growth factor - release, and CB2 antagonism on Tregs impairs their ability to suppress T cells. In our second aim, MGL was investigated in primary human epithelial esophageal cells and primary human lung fibroblasts by qPCR, and we applied CRISPR/Cas9 gene knockout, performed proliferation assays and Western blots, and showed that MGL is expressed by epithelial cells and that MGL knockout increases the proliferation of fibroblasts. Also, it was found that IL-13 could function as a positive control for fibroblast proliferation. In conclusion, the results of this thesis indicate that the endocannabinoid system is involved in the pathology of eosinophilic esophagitis. Thus, the CB2 receptor as well as other components of the endocannabinoid system in EoE should be further investigated.