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A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration
PURPOSE: The RPE cells have a major role in the development of dry age-related macular degeneration (AMD). We present novel evidence that βA3/A1-crystallin, encoded by the Cryba1 gene, a protein known to be important for lysosomal clearance in the RPE, also has a role in epithelial-to-mesenchymal tr...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Association for Research in Vision and Ophthalmology
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058694/ https://www.ncbi.nlm.nih.gov/pubmed/30098172 http://dx.doi.org/10.1167/iovs.18-24132 |
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author | Ghosh, Sayan Shang, Peng Terasaki, Hiroto Stepicheva, Nadezda Hose, Stacey Yazdankhah, Meysam Weiss, Joseph Sakamoto, Taiji Bhutto, Imran A. Xia, Shuli Zigler, J. Samuel Kannan, Ram Qian, Jiang Handa, James T. Sinha, Debasish |
author_facet | Ghosh, Sayan Shang, Peng Terasaki, Hiroto Stepicheva, Nadezda Hose, Stacey Yazdankhah, Meysam Weiss, Joseph Sakamoto, Taiji Bhutto, Imran A. Xia, Shuli Zigler, J. Samuel Kannan, Ram Qian, Jiang Handa, James T. Sinha, Debasish |
author_sort | Ghosh, Sayan |
collection | PubMed |
description | PURPOSE: The RPE cells have a major role in the development of dry age-related macular degeneration (AMD). We present novel evidence that βA3/A1-crystallin, encoded by the Cryba1 gene, a protein known to be important for lysosomal clearance in the RPE, also has a role in epithelial-to-mesenchymal transition (EMT) of RPE cells. METHODS: RPE from dry AMD globes, genetically engineered mice lacking Cryba1 globally or specifically in the RPE, spontaneous mutant rats (Nuc1) with a loss-of-function mutation in Cryba1, and the melanoma OCM3 cell line were used. Spatial localization of proteins was demonstrated with immunofluorescence, gene expression levels were determined by quantitative PCR (qPCR), and protein levels by Western blotting. Cell movement was evaluated using wound healing and cell migration assays. Co-immunoprecipitation was used to identify binding partners of βA3/A1-crystallin. RESULTS: βA3/A1-crystallin is upregulated in polarized RPE cells compared to undifferentiated cells. Loss of βA3/A1-crystallin in murine and human RPE cells resulted in upregulation of Snail and vimentin, downregulation of E-cadherin, and increased cell migration. βA3/A1-crystallin binds to cortactin, and loss of βA3/A1-crystallin resulted in increased P-cortactin(Y421). The RPE from AMD samples had increased Snail and vimentin, and decreased E-cadherin, compared to age-matched controls. CONCLUSIONS: We introduced a novel concept of dry AMD initiation induced by lysosomal clearance defects in the RPE and subsequent attempts by RPE cells to avoid the resulting stress by undergoing EMT. We demonstrate that βA3/A1-crystallin is a potential therapeutic target for AMD through rejuvenation of lysosomal dysfunction and potentially, reversal of EMT. |
format | Online Article Text |
id | pubmed-6058694 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The Association for Research in Vision and Ophthalmology |
record_format | MEDLINE/PubMed |
spelling | pubmed-60586942018-07-26 A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration Ghosh, Sayan Shang, Peng Terasaki, Hiroto Stepicheva, Nadezda Hose, Stacey Yazdankhah, Meysam Weiss, Joseph Sakamoto, Taiji Bhutto, Imran A. Xia, Shuli Zigler, J. Samuel Kannan, Ram Qian, Jiang Handa, James T. Sinha, Debasish Invest Ophthalmol Vis Sci Special Issue PURPOSE: The RPE cells have a major role in the development of dry age-related macular degeneration (AMD). We present novel evidence that βA3/A1-crystallin, encoded by the Cryba1 gene, a protein known to be important for lysosomal clearance in the RPE, also has a role in epithelial-to-mesenchymal transition (EMT) of RPE cells. METHODS: RPE from dry AMD globes, genetically engineered mice lacking Cryba1 globally or specifically in the RPE, spontaneous mutant rats (Nuc1) with a loss-of-function mutation in Cryba1, and the melanoma OCM3 cell line were used. Spatial localization of proteins was demonstrated with immunofluorescence, gene expression levels were determined by quantitative PCR (qPCR), and protein levels by Western blotting. Cell movement was evaluated using wound healing and cell migration assays. Co-immunoprecipitation was used to identify binding partners of βA3/A1-crystallin. RESULTS: βA3/A1-crystallin is upregulated in polarized RPE cells compared to undifferentiated cells. Loss of βA3/A1-crystallin in murine and human RPE cells resulted in upregulation of Snail and vimentin, downregulation of E-cadherin, and increased cell migration. βA3/A1-crystallin binds to cortactin, and loss of βA3/A1-crystallin resulted in increased P-cortactin(Y421). The RPE from AMD samples had increased Snail and vimentin, and decreased E-cadherin, compared to age-matched controls. CONCLUSIONS: We introduced a novel concept of dry AMD initiation induced by lysosomal clearance defects in the RPE and subsequent attempts by RPE cells to avoid the resulting stress by undergoing EMT. We demonstrate that βA3/A1-crystallin is a potential therapeutic target for AMD through rejuvenation of lysosomal dysfunction and potentially, reversal of EMT. The Association for Research in Vision and Ophthalmology 2018-07 /pmc/articles/PMC6058694/ /pubmed/30098172 http://dx.doi.org/10.1167/iovs.18-24132 Text en Copyright 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. |
spellingShingle | Special Issue Ghosh, Sayan Shang, Peng Terasaki, Hiroto Stepicheva, Nadezda Hose, Stacey Yazdankhah, Meysam Weiss, Joseph Sakamoto, Taiji Bhutto, Imran A. Xia, Shuli Zigler, J. Samuel Kannan, Ram Qian, Jiang Handa, James T. Sinha, Debasish A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title | A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title_full | A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title_fullStr | A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title_full_unstemmed | A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title_short | A Role for βA3/A1-Crystallin in Type 2 EMT of RPE Cells Occurring in Dry Age-Related Macular Degeneration |
title_sort | role for βa3/a1-crystallin in type 2 emt of rpe cells occurring in dry age-related macular degeneration |
topic | Special Issue |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6058694/ https://www.ncbi.nlm.nih.gov/pubmed/30098172 http://dx.doi.org/10.1167/iovs.18-24132 |
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