The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment
Primary open-angle glaucoma (POAG) constitutes the most common type of glaucoma. Emerging evidence suggests that Endoplasmic Reticulum (ER) stress and the protein kinase RNA-like endoplasmic reticulum kinase (PERK)-mediated Unfolded Protein Response (UPR) signaling pathway play a key role in POAG pa...
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MDPI AG
2021-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/9/4494 |
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| author | Wioletta Rozpędek-Kamińska Grzegorz Galita Natalia Siwecka Steven L. Carroll John Alan Diehl Ewa Kucharska Dariusz Pytel Ireneusz Majsterek |
| author_facet | Wioletta Rozpędek-Kamińska Grzegorz Galita Natalia Siwecka Steven L. Carroll John Alan Diehl Ewa Kucharska Dariusz Pytel Ireneusz Majsterek |
| author_sort | Wioletta Rozpędek-Kamińska |
| collection | DOAJ |
| description | Primary open-angle glaucoma (POAG) constitutes the most common type of glaucoma. Emerging evidence suggests that Endoplasmic Reticulum (ER) stress and the protein kinase RNA-like endoplasmic reticulum kinase (PERK)-mediated Unfolded Protein Response (UPR) signaling pathway play a key role in POAG pathogenesis. Thus, the main aim of the study was to evaluate the effectiveness of the PERK inhibitor LDN-0060609 in cellular model of glaucoma using primary human trabecular meshwork (HTM) cells. To evaluate the level of the ER stress marker proteins, Western blotting and TaqMan gene expression assay were used. The cytotoxicity was measured by XTT, LDH assays and Giemsa staining, whereas genotoxicity via comet assay. Changes in cell morphology were assessed by phase-contrast microscopy. Analysis of apoptosis was performed by caspase-3 assay and flow cytometry (FC), whereas cell cycle progression by FC. The results obtained have demonstrated that LDN-0060609 triggered a significant decrease of ER stress marker proteins within HTM cells with induced ER stress conditions. Moreover, LDN-0060609 effectively increased viability, reduced DNA damage, increased proliferation, restored normal morphology, reduced apoptosis and restored normal cell cycle distribution of HTM cells with induced ER stress conditions. Thereby, PERK inhibitors, such as LDN-0060609, may provide an innovative, ground-breaking treatment strategy against POAG. |
| first_indexed | 2024-03-10T11:58:18Z |
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| id | doaj.art-c62578b9a289483f9d5902fe21f78491 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T11:58:18Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-c62578b9a289483f9d5902fe21f784912023-11-21T17:09:47ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-04-01229449410.3390/ijms22094494The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma TreatmentWioletta Rozpędek-Kamińska0Grzegorz Galita1Natalia Siwecka2Steven L. Carroll3John Alan Diehl4Ewa Kucharska5Dariusz Pytel6Ireneusz Majsterek7Department of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandDepartment of Pathology and Laboratory Medicine, Medical University of South Carolina, Charleston, SC 29425, USAHollings Cancer Center, Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Charleston, SC 29425, USADepartment of Gerontology, Geriatrics and Social Work, Jesuit University Ignatianum, 31-501 Krakow, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandDepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandPrimary open-angle glaucoma (POAG) constitutes the most common type of glaucoma. Emerging evidence suggests that Endoplasmic Reticulum (ER) stress and the protein kinase RNA-like endoplasmic reticulum kinase (PERK)-mediated Unfolded Protein Response (UPR) signaling pathway play a key role in POAG pathogenesis. Thus, the main aim of the study was to evaluate the effectiveness of the PERK inhibitor LDN-0060609 in cellular model of glaucoma using primary human trabecular meshwork (HTM) cells. To evaluate the level of the ER stress marker proteins, Western blotting and TaqMan gene expression assay were used. The cytotoxicity was measured by XTT, LDH assays and Giemsa staining, whereas genotoxicity via comet assay. Changes in cell morphology were assessed by phase-contrast microscopy. Analysis of apoptosis was performed by caspase-3 assay and flow cytometry (FC), whereas cell cycle progression by FC. The results obtained have demonstrated that LDN-0060609 triggered a significant decrease of ER stress marker proteins within HTM cells with induced ER stress conditions. Moreover, LDN-0060609 effectively increased viability, reduced DNA damage, increased proliferation, restored normal morphology, reduced apoptosis and restored normal cell cycle distribution of HTM cells with induced ER stress conditions. Thereby, PERK inhibitors, such as LDN-0060609, may provide an innovative, ground-breaking treatment strategy against POAG.https://www.mdpi.com/1422-0067/22/9/4494glaucomaPERKeIF2αendoplasmic reticulum stressunfolded protein responseapoptosis |
| spellingShingle | Wioletta Rozpędek-Kamińska Grzegorz Galita Natalia Siwecka Steven L. Carroll John Alan Diehl Ewa Kucharska Dariusz Pytel Ireneusz Majsterek The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment International Journal of Molecular Sciences glaucoma PERK eIF2α endoplasmic reticulum stress unfolded protein response apoptosis |
| title | The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment |
| title_full | The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment |
| title_fullStr | The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment |
| title_full_unstemmed | The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment |
| title_short | The Potential Role of Small-Molecule PERK Inhibitor LDN-0060609 in Primary Open-Angle Glaucoma Treatment |
| title_sort | potential role of small molecule perk inhibitor ldn 0060609 in primary open angle glaucoma treatment |
| topic | glaucoma PERK eIF2α endoplasmic reticulum stress unfolded protein response apoptosis |
| url | https://www.mdpi.com/1422-0067/22/9/4494 |
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