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- Title
Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach.
- Authors
Lipski, Deborah A.; Foucart, Vincent; Dewispelaere, Rémi; Caspers, Laure E.; Defrance, Matthieu; Bruyns, Catherine; Willermain, François
- Abstract
<bold>Background: </bold>Blood-retinal barrier cells are known to exhibit a massive phenotypic change during experimental autoimmune uveitis (EAU) development. In an attempt to investigate the mechanisms of blood-retinal barrier (BRB) breakdown at a global level, we studied the gene regulation of total retinal cells and retinal endothelial cells during non-infectious uveitis.<bold>Methods: </bold>Retinal endothelial cells were isolated by flow cytometry either in Tie2-GFP mice (CD31+ CD45- GFP+ cells), or in wild type C57BL/6 mice (CD31+ CD45- endoglin+ cells). EAU was induced in C57BL/6 mice by adoptive transfer of IRBP1-20-specific T cells. Total retinal cells and retinal endothelial cells from naïve and EAU mice were sorted and their gene expression compared by RNA-Seq. Protein expression of selected genes was validated by immunofluorescence on retinal wholemounts and cryosections and by flow cytometry.<bold>Results: </bold>Retinal endothelial cell sorting in wild type C57BL/6 mice was validated by comparative transcriptome analysis with retinal endothelial cells sorted from Tie2-GFP mice, which express GFP under the control of the endothelial-specific receptor tyrosine kinase promoter Tie2. RNA-Seq analysis of total retinal cells mainly brought to light upregulation of genes involved in antigen presentation and T cell activation during EAU. Specific transcriptome analysis of retinal endothelial cells allowed us to identify 82 genes modulated in retinal endothelial cells during EAU development. Protein expression of 5 of those genes (serpina3n, lcn2, ackr1, lrg1 and lamc3) was validated at the level of inner BRB cells.<bold>Conclusion: </bold>Those data not only confirm the involvement of known pathogenic molecules but further provide a list of new candidate genes and pathways possibly implicated in inner BRB breakdown during non-infectious posterior uveitis.
- Subjects
UGANDA; ENDOTHELIAL cells; UVEITIS; GENETIC regulation; PROTEIN-tyrosine kinases; T cells
- Publication
BMC Ophthalmology, 2020, Vol 20, Issue 1, p1
- ISSN
1471-2415
- Publication type
journal article
- DOI
10.1186/s12886-020-1333-5