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YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction
PURPOSE: Elevated transforming growth factor beta2 (TGFβ2) levels in the aqueous humor have been linked to glaucomatous outflow tissue dysfunction. Potential mediators of dysfunction are the transcriptional coactivators, Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding m...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Association for Research in Vision and Ophthalmology
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9652721/ https://www.ncbi.nlm.nih.gov/pubmed/36350617 http://dx.doi.org/10.1167/iovs.63.12.15 |
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author | Li, Haiyan Singh, Ayushi Perkumas, Kristin M. Stamer, W. Daniel Ganapathy, Preethi S. Herberg, Samuel |
author_facet | Li, Haiyan Singh, Ayushi Perkumas, Kristin M. Stamer, W. Daniel Ganapathy, Preethi S. Herberg, Samuel |
author_sort | Li, Haiyan |
collection | PubMed |
description | PURPOSE: Elevated transforming growth factor beta2 (TGFβ2) levels in the aqueous humor have been linked to glaucomatous outflow tissue dysfunction. Potential mediators of dysfunction are the transcriptional coactivators, Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding motif (TAZ). However, the molecular underpinnings of YAP/TAZ modulation in Schlemm's canal (SC) cells under glaucomatous conditions are not well understood. Here, we investigate how TGFβ2 regulates YAP/TAZ activity in human SC (HSC) cells using biomimetic extracellular matrix hydrogels, and examine whether pharmacological YAP/TAZ inhibition would attenuate TGFβ2-induced HSC cell dysfunction. METHODS: Primary HSC cells were seeded atop photo-cross-linked extracellular matrix hydrogels, made of collagen type I, elastin-like polypeptide and hyaluronic acid, or encapsulated within the hydrogels. HSC cells were induced with TGFβ2 in the absence or presence of concurrent actin destabilization or pharmacological YAP/TAZ inhibition. Changes in actin cytoskeletal organization, YAP/TAZ activity, extracellular matrix production, phospho-myosin light chain levels, and hydrogel contraction were assessed. RESULTS: TGFβ2 significantly increased YAP/TAZ nuclear localization in HSC cells, which was prevented by either filamentous-actin relaxation or depolymerization. Pharmacological YAP/TAZ inhibition using verteporfin without light stimulation decreased fibronectin expression and actomyosin cytoskeletal rearrangement in HSC cells induced by TGFβ2. Similarly, verteporfin significantly attenuated TGFβ2-induced HSC cell-encapsulated hydrogel contraction. CONCLUSIONS: Our data provide evidence for a pathologic role of aberrant YAP/TAZ signaling in HSC cells under simulated glaucomatous conditions and suggest that pharmacological YAP/TAZ inhibition has promising potential to improve outflow tissue dysfunction. |
format | Online Article Text |
id | pubmed-9652721 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Association for Research in Vision and Ophthalmology |
record_format | MEDLINE/PubMed |
spelling | pubmed-96527212022-11-15 YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction Li, Haiyan Singh, Ayushi Perkumas, Kristin M. Stamer, W. Daniel Ganapathy, Preethi S. Herberg, Samuel Invest Ophthalmol Vis Sci Glaucoma PURPOSE: Elevated transforming growth factor beta2 (TGFβ2) levels in the aqueous humor have been linked to glaucomatous outflow tissue dysfunction. Potential mediators of dysfunction are the transcriptional coactivators, Yes-associated protein (YAP) and transcriptional coactivator with PDZ binding motif (TAZ). However, the molecular underpinnings of YAP/TAZ modulation in Schlemm's canal (SC) cells under glaucomatous conditions are not well understood. Here, we investigate how TGFβ2 regulates YAP/TAZ activity in human SC (HSC) cells using biomimetic extracellular matrix hydrogels, and examine whether pharmacological YAP/TAZ inhibition would attenuate TGFβ2-induced HSC cell dysfunction. METHODS: Primary HSC cells were seeded atop photo-cross-linked extracellular matrix hydrogels, made of collagen type I, elastin-like polypeptide and hyaluronic acid, or encapsulated within the hydrogels. HSC cells were induced with TGFβ2 in the absence or presence of concurrent actin destabilization or pharmacological YAP/TAZ inhibition. Changes in actin cytoskeletal organization, YAP/TAZ activity, extracellular matrix production, phospho-myosin light chain levels, and hydrogel contraction were assessed. RESULTS: TGFβ2 significantly increased YAP/TAZ nuclear localization in HSC cells, which was prevented by either filamentous-actin relaxation or depolymerization. Pharmacological YAP/TAZ inhibition using verteporfin without light stimulation decreased fibronectin expression and actomyosin cytoskeletal rearrangement in HSC cells induced by TGFβ2. Similarly, verteporfin significantly attenuated TGFβ2-induced HSC cell-encapsulated hydrogel contraction. CONCLUSIONS: Our data provide evidence for a pathologic role of aberrant YAP/TAZ signaling in HSC cells under simulated glaucomatous conditions and suggest that pharmacological YAP/TAZ inhibition has promising potential to improve outflow tissue dysfunction. The Association for Research in Vision and Ophthalmology 2022-11-09 /pmc/articles/PMC9652721/ /pubmed/36350617 http://dx.doi.org/10.1167/iovs.63.12.15 Text en Copyright 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. |
spellingShingle | Glaucoma Li, Haiyan Singh, Ayushi Perkumas, Kristin M. Stamer, W. Daniel Ganapathy, Preethi S. Herberg, Samuel YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title | YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title_full | YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title_fullStr | YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title_full_unstemmed | YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title_short | YAP/TAZ Mediate TGFβ2-Induced Schlemm's Canal Cell Dysfunction |
title_sort | yap/taz mediate tgfβ2-induced schlemm's canal cell dysfunction |
topic | Glaucoma |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9652721/ https://www.ncbi.nlm.nih.gov/pubmed/36350617 http://dx.doi.org/10.1167/iovs.63.12.15 |
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