Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells
Clinical use of glucocorticoids is associated with increased intraocular pressure (IOP), a major risk factor for glaucoma. Glucocorticoids have been reported to induce changes in actin cytoskeletal organization, cell adhesion, extracellular matrix, fibrogenic activity, and mechanical properties of t...
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Frontiers Media S.A.
2022-04-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2022.886754/full |
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| author | William Bachman Rupalatha Maddala Ayon Chakraborty Camelia Eldawy Nikolai P. Skiba Ponugoti V. Rao Ponugoti V. Rao |
| author_facet | William Bachman Rupalatha Maddala Ayon Chakraborty Camelia Eldawy Nikolai P. Skiba Ponugoti V. Rao Ponugoti V. Rao |
| author_sort | William Bachman |
| collection | DOAJ |
| description | Clinical use of glucocorticoids is associated with increased intraocular pressure (IOP), a major risk factor for glaucoma. Glucocorticoids have been reported to induce changes in actin cytoskeletal organization, cell adhesion, extracellular matrix, fibrogenic activity, and mechanical properties of trabecular meshwork (TM) tissue, which plays a crucial role in aqueous humor dynamics and IOP homeostasis. However, we have a limited understanding of the molecular underpinnings regulating these myriad processes in TM cells. To understand how proteins, including cytoskeletal and cell adhesion proteins that are recognized to shuttle between the cytosolic and nuclear regions, influence gene expression and other cellular activities, we used proteomic analysis to characterize the nuclear protein fraction of dexamethasone (Dex) treated human TM cells. Treatment of human TM cells with Dex for 1, 5, or 7 days led to consistent increases (by ≥ two-fold) in the levels of various actin cytoskeletal regulatory, cell adhesive, and vesicle trafficking proteins. Increases (≥two-fold) were also observed in levels of Wnt signaling regulator (glypican-4), actin-binding chromatin modulator (BRG1) and nuclear actin filament depolymerizing protein (MICAL2; microtubule-associated monooxygenase, calponin and LIM domain containing), together with a decrease in tissue plasminogen activator. These changes were independently further confirmed by immunoblotting analysis. Interestingly, deficiency of BRG1 expression blunted the Dex-induced increases in the levels of some of these proteins in TM cells. In summary, these findings indicate that the widely recognized changes in actin cytoskeletal and cell adhesive attributes of TM cells by glucocorticoids involve actin regulated BRG1 chromatin remodeling, nuclear MICAL2, and glypican-4 regulated Wnt signaling upstream of the serum response factor/myocardin controlled transcriptional activity. |
| first_indexed | 2024-12-12T22:26:04Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-12-12T22:26:04Z |
| publishDate | 2022-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-7016b0f6432046b8b147f4272e01cdad2022-12-22T00:09:45ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2022-04-011010.3389/fcell.2022.886754886754Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork CellsWilliam Bachman0Rupalatha Maddala1Ayon Chakraborty2Camelia Eldawy3Nikolai P. Skiba4Ponugoti V. Rao5Ponugoti V. Rao6Department of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Ophthalmology, Duke University School of Medicine, Durham, NC, United StatesDepartment of Pharmacology and Cancer Biology, Duke University School of Medicine, Durham, NC, United StatesClinical use of glucocorticoids is associated with increased intraocular pressure (IOP), a major risk factor for glaucoma. Glucocorticoids have been reported to induce changes in actin cytoskeletal organization, cell adhesion, extracellular matrix, fibrogenic activity, and mechanical properties of trabecular meshwork (TM) tissue, which plays a crucial role in aqueous humor dynamics and IOP homeostasis. However, we have a limited understanding of the molecular underpinnings regulating these myriad processes in TM cells. To understand how proteins, including cytoskeletal and cell adhesion proteins that are recognized to shuttle between the cytosolic and nuclear regions, influence gene expression and other cellular activities, we used proteomic analysis to characterize the nuclear protein fraction of dexamethasone (Dex) treated human TM cells. Treatment of human TM cells with Dex for 1, 5, or 7 days led to consistent increases (by ≥ two-fold) in the levels of various actin cytoskeletal regulatory, cell adhesive, and vesicle trafficking proteins. Increases (≥two-fold) were also observed in levels of Wnt signaling regulator (glypican-4), actin-binding chromatin modulator (BRG1) and nuclear actin filament depolymerizing protein (MICAL2; microtubule-associated monooxygenase, calponin and LIM domain containing), together with a decrease in tissue plasminogen activator. These changes were independently further confirmed by immunoblotting analysis. Interestingly, deficiency of BRG1 expression blunted the Dex-induced increases in the levels of some of these proteins in TM cells. In summary, these findings indicate that the widely recognized changes in actin cytoskeletal and cell adhesive attributes of TM cells by glucocorticoids involve actin regulated BRG1 chromatin remodeling, nuclear MICAL2, and glypican-4 regulated Wnt signaling upstream of the serum response factor/myocardin controlled transcriptional activity.https://www.frontiersin.org/articles/10.3389/fcell.2022.886754/fullglaucomaintraocular pressureglucocorticoidstrabecular meshworknucleoskeletonproteomics |
| spellingShingle | William Bachman Rupalatha Maddala Ayon Chakraborty Camelia Eldawy Nikolai P. Skiba Ponugoti V. Rao Ponugoti V. Rao Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells Frontiers in Cell and Developmental Biology glaucoma intraocular pressure glucocorticoids trabecular meshwork nucleoskeleton proteomics |
| title | Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells |
| title_full | Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells |
| title_fullStr | Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells |
| title_full_unstemmed | Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells |
| title_short | Glucocorticoids Preferentially Influence Expression of Nucleoskeletal Actin Network and Cell Adhesive Proteins in Human Trabecular Meshwork Cells |
| title_sort | glucocorticoids preferentially influence expression of nucleoskeletal actin network and cell adhesive proteins in human trabecular meshwork cells |
| topic | glaucoma intraocular pressure glucocorticoids trabecular meshwork nucleoskeleton proteomics |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2022.886754/full |
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