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| Status |
Public on Jan 28, 2020 |
| Title |
Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Background: 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. Methods: 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. Results: 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, lipocalin 2, ackr1, lrg1 and lamc3) was validated at the level of inner BRB cells. Conclusion: 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.
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| Overall design |
Regulation of retinal cell and retinal endothelial cell gene expression during experimental autoimmune uveitis
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| Contributor(s) |
Lipski DA, Foucart V, Dewispelaere R, Caspers LE, Defrance M, Bruyns C, Willermain F |
| Citation(s) |
32183784 |
| Submission date |
Jan 23, 2020 |
| Last update date |
Mar 24, 2020 |
| Contact name |
Frederick Libert |
| E-mail(s) |
flibert@ulb.ac.be
|
| Organization name |
ULB
|
| Department |
IRIBHM
|
| Lab |
C2-140
|
| Street address |
808 route de Lennik
|
| City |
Brussels |
| ZIP/Postal code |
B-1070 |
| Country |
Belgium |
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| Platforms (1) |
| GPL18480 |
Illumina HiSeq 1500 (Mus musculus) |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA602922 |
| SRA |
SRP244473 |
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