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Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach
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 c...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7076950/ https://www.ncbi.nlm.nih.gov/pubmed/32183784 http://dx.doi.org/10.1186/s12886-020-1333-5 |
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author | Lipski, Deborah A. Foucart, Vincent Dewispelaere, Rémi Caspers, Laure E. Defrance, Matthieu Bruyns, Catherine Willermain, François |
author_facet | Lipski, Deborah A. Foucart, Vincent Dewispelaere, Rémi Caspers, Laure E. Defrance, Matthieu Bruyns, Catherine Willermain, François |
author_sort | Lipski, Deborah A. |
collection | PubMed |
description | 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, lcn2, 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. |
format | Online Article Text |
id | pubmed-7076950 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-70769502020-03-18 Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach Lipski, Deborah A. Foucart, Vincent Dewispelaere, Rémi Caspers, Laure E. Defrance, Matthieu Bruyns, Catherine Willermain, François BMC Ophthalmol Research Article 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, lcn2, 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. BioMed Central 2020-03-17 /pmc/articles/PMC7076950/ /pubmed/32183784 http://dx.doi.org/10.1186/s12886-020-1333-5 Text en © The Author(s). 2020 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Article Lipski, Deborah A. Foucart, Vincent Dewispelaere, Rémi Caspers, Laure E. Defrance, Matthieu Bruyns, Catherine Willermain, François Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title | Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title_full | Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title_fullStr | Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title_full_unstemmed | Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title_short | Retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
title_sort | retinal endothelial cell phenotypic modifications during experimental autoimmune uveitis: a transcriptomic approach |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7076950/ https://www.ncbi.nlm.nih.gov/pubmed/32183784 http://dx.doi.org/10.1186/s12886-020-1333-5 |
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