Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes
The term glaucoma encompasses various neurodegenerative eye disorders, among which the most common is primary open-angle glaucoma (POAG). Recently, the essential role of human retinal astrocytes (HRA) in glaucoma progression has been placed in the spotlight. It has been found that placing the endopl...
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| Format: | Article |
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MDPI AG
2024-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/25/2/728 |
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| author | Wioletta Rozpędek-Kamińska Grzegorz Galita Kamil Saramowicz Zuzanna Granek Julia Barczuk Natalia Siwecka Dariusz Pytel Ireneusz Majsterek |
| author_facet | Wioletta Rozpędek-Kamińska Grzegorz Galita Kamil Saramowicz Zuzanna Granek Julia Barczuk Natalia Siwecka Dariusz Pytel Ireneusz Majsterek |
| author_sort | Wioletta Rozpędek-Kamińska |
| collection | DOAJ |
| description | The term glaucoma encompasses various neurodegenerative eye disorders, among which the most common is primary open-angle glaucoma (POAG). Recently, the essential role of human retinal astrocytes (HRA) in glaucoma progression has been placed in the spotlight. It has been found that placing the endoplasmic reticulum (ER) under stress and activating PERK leads to apoptosis of HRA cells, which inhibits their neuroprotective effect in the course of glaucoma. Therefore, the aim of the present study was to evaluate the effectiveness of the small-molecule PERK inhibitor LDN-0060609 in countering ER stress conditions induced in HRA cells in vitro. The activity of LDN-0060609 was studied in terms of protein and mRNA expression, cytotoxicity, genotoxicity, caspase-3 level and cell cycle progression. LDN-0060609 at 25 μM proved to be a potent inhibitor of the major PERK substrate, p-eIF2α (49% inhibition). The compound markedly decreased the expression of pro-apoptotic ER stress-related genes (<i>ATF4</i>, <i>DDIT3</i>, <i>BAX</i> and <i>Bcl-2</i>). Treatment with LDN-0060609 significantly increased cell viability, decreased genotoxicity and caspase-3 levels, and restored cell cycle distribution in HRA cells with activated ER stress conditions. These findings indicate that the small-molecule PERK inhibitor LDN-0060609 can potentially be developed into a novel anti-glaucoma agent. |
| first_indexed | 2024-03-08T09:55:27Z |
| format | Article |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-08T09:55:27Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-fd3f2d429343417993b7495edfebb6212024-01-29T13:53:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-01-0125272810.3390/ijms25020728Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal AstrocytesWioletta Rozpędek-Kamińska0Grzegorz Galita1Kamil Saramowicz2Zuzanna Granek3Julia Barczuk4Natalia Siwecka5Dariusz 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 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, USADepartment of Clinical Chemistry and Biochemistry, Medical University of Lodz, 90-419 Lodz, PolandThe term glaucoma encompasses various neurodegenerative eye disorders, among which the most common is primary open-angle glaucoma (POAG). Recently, the essential role of human retinal astrocytes (HRA) in glaucoma progression has been placed in the spotlight. It has been found that placing the endoplasmic reticulum (ER) under stress and activating PERK leads to apoptosis of HRA cells, which inhibits their neuroprotective effect in the course of glaucoma. Therefore, the aim of the present study was to evaluate the effectiveness of the small-molecule PERK inhibitor LDN-0060609 in countering ER stress conditions induced in HRA cells in vitro. The activity of LDN-0060609 was studied in terms of protein and mRNA expression, cytotoxicity, genotoxicity, caspase-3 level and cell cycle progression. LDN-0060609 at 25 μM proved to be a potent inhibitor of the major PERK substrate, p-eIF2α (49% inhibition). The compound markedly decreased the expression of pro-apoptotic ER stress-related genes (<i>ATF4</i>, <i>DDIT3</i>, <i>BAX</i> and <i>Bcl-2</i>). Treatment with LDN-0060609 significantly increased cell viability, decreased genotoxicity and caspase-3 levels, and restored cell cycle distribution in HRA cells with activated ER stress conditions. These findings indicate that the small-molecule PERK inhibitor LDN-0060609 can potentially be developed into a novel anti-glaucoma agent.https://www.mdpi.com/1422-0067/25/2/728eIF2αendoplasmic reticulum stressglaucomaglaucoma treatmentPERKPERK inhibitor |
| spellingShingle | Wioletta Rozpędek-Kamińska Grzegorz Galita Kamil Saramowicz Zuzanna Granek Julia Barczuk Natalia Siwecka Dariusz Pytel Ireneusz Majsterek Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes International Journal of Molecular Sciences eIF2α endoplasmic reticulum stress glaucoma glaucoma treatment PERK PERK inhibitor |
| title | Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes |
| title_full | Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes |
| title_fullStr | Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes |
| title_full_unstemmed | Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes |
| title_short | Evaluation of the LDN-0060609 PERK Inhibitor as a Selective Treatment for Primary Open-Angle Glaucoma: An In Vitro Study on Human Retinal Astrocytes |
| title_sort | evaluation of the ldn 0060609 perk inhibitor as a selective treatment for primary open angle glaucoma an in vitro study on human retinal astrocytes |
| topic | eIF2α endoplasmic reticulum stress glaucoma glaucoma treatment PERK PERK inhibitor |
| url | https://www.mdpi.com/1422-0067/25/2/728 |
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