Abstract
Purpose :
Experimental autoimmune uveitis (EAU) is the most common rodent model of autoimmune uveitis.
To gain more understanding of the underlying pathology of EAU, analyzing high-throughput molecular measurements at a functional level is important. Especially, the knowledge base-driven pathway analysis is a well-established method, which mainly exploit pathway knowledge in public repositories, e.g. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG).
In this study, we used RNA-sequencing to map DEGs and identify key signaling pathways of EAU.
Methods :
EAU was induced in 16 female B10.RIII mice by Inter-Photoreceptor-Binding-Protein (IRBP) in complete Freund's adjuvant, boosted by Pertussis toxin. 4 mice each were euthanized and enucleated on day 7, 10, 13 and 18 respectively. 4 mice without EAU served as control group. The retinas were dissected and snap frozen. RNA was isolated and RNA sequencing was performed. Up- and down-regulated genes were selected after pair-wise analysis with the control group. To gain more insights into functional levels of DEGs, GO and KEGG analysis were carried out.
Results :
In KEGG 4 of the 58 up-regulated genes play a role in phototransduction. Clusters of down-regulated genes (in total 132) contained several transcripts that had been previously related to focal adhesion, Wnt signaling pathway, and TGF-beta signaling pathway.
We mapped the distribution of DEGs in different GO functions, namely the biological process (BP), cell component (CC) and molecular function (MF). Examples of BP of up-regulated genes involved positive regulation of cytokine production and cytokine-mediated signaling pathways. Examples of BP of down-regulated genes involved eye development and visual system development. In MF differentiation, cytokine receptor activity and cytokine receptor binding were among the up-regulates DEGs. RNA polymerase binding and histone binding were among the down-regulated DEGs.
Conclusions :
In EAU mice we found a strong alteration of the transcript signature compared to non-EAU retinas. A clear trace of inflammatory pathways was found in clustering the up-regulated DEGs. In contrast, down-regulated DEGs involved processes of visual development. These data sets suggest a number of pathways, which could serve as potential immunoregulatory targets in the understanding and finally in the prevention and treatment of autoimmune uveitis.
This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.