GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia
Apoptotic cell death is a deleterious consequence of hypoxia-induced cellular stress. The master <i>hypoxamiR</i>, microRNA-210 (miR-210), is considered the primary driver of the cellular response to hypoxia stress. We have recently demonstrated that miR-210 attenuates hypoxia-induced ap...
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MDPI AG
2022-08-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/16/9375 |
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| author | Gurdeep Marwarha Øystein Røsand Katrine Hordnes Slagsvold Morten Andre Høydal |
| author_facet | Gurdeep Marwarha Øystein Røsand Katrine Hordnes Slagsvold Morten Andre Høydal |
| author_sort | Gurdeep Marwarha |
| collection | DOAJ |
| description | Apoptotic cell death is a deleterious consequence of hypoxia-induced cellular stress. The master <i>hypoxamiR</i>, microRNA-210 (miR-210), is considered the primary driver of the cellular response to hypoxia stress. We have recently demonstrated that miR-210 attenuates hypoxia-induced apoptotic cell death. In this paper, we unveil that the miR-210-induced inhibition of the serine/threonine kinase Glycogen Synthase Kinase 3 beta (GSK3β) in AC-16 cardiomyocytes subjected to hypoxia stress underlies the salutary protective response of miR-210 in mitigating the hypoxia-induced apoptotic cell death. Using transient overexpression vectors to augment miR-210 expression concomitant with the ectopic expression of the <i>constitutive active</i> GSK3β S9A mutant (<i>ca</i>-GSK3β S9A), we exhaustively performed biochemical and molecular assays to determine the status of the hypoxia-induced intrinsic apoptosis cascade. Caspase-3 activity analysis coupled with DNA fragmentation assays cogently demonstrate that the inhibition of GSK3β kinase activity underlies the miR-210-induced attenuation in the hypoxia-driven apoptotic cell death. Further elucidation and delineation of the upstream cellular events unveiled an indispensable role of the inhibition of GSK3β kinase activity in mediating the miR-210-induced mitigation of the hypoxia-driven BAX and BAK insertion into the <i>outer mitochondria membrane</i> (<i>OMM</i>) and the ensuing <i>Cytochrome C</i> release into the cytosol. Our study is the first to unveil that the inhibition of GSK3β kinase activity is indispensable in mediating the miR-210-orchestrated protective cellular response to hypoxia-induced apoptotic cell death. |
| first_indexed | 2024-03-09T13:15:43Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T13:15:43Z |
| publishDate | 2022-08-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-a3b834db9b8d401da86ad84f1602ac072023-11-30T21:36:35ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-08-012316937510.3390/ijms23169375GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during HypoxiaGurdeep Marwarha0Øystein Røsand1Katrine Hordnes Slagsvold2Morten Andre Høydal3Group of Molecular and Cellular Cardiology, Department of Circulation and Medical Imaging, Faculty of Medicine and Health, Norwegian University of Science and Technology (NTNU), 7034 Trondheim, NorwayGroup of Molecular and Cellular Cardiology, Department of Circulation and Medical Imaging, Faculty of Medicine and Health, Norwegian University of Science and Technology (NTNU), 7034 Trondheim, NorwayGroup of Molecular and Cellular Cardiology, Department of Circulation and Medical Imaging, Faculty of Medicine and Health, Norwegian University of Science and Technology (NTNU), 7034 Trondheim, NorwayGroup of Molecular and Cellular Cardiology, Department of Circulation and Medical Imaging, Faculty of Medicine and Health, Norwegian University of Science and Technology (NTNU), 7034 Trondheim, NorwayApoptotic cell death is a deleterious consequence of hypoxia-induced cellular stress. The master <i>hypoxamiR</i>, microRNA-210 (miR-210), is considered the primary driver of the cellular response to hypoxia stress. We have recently demonstrated that miR-210 attenuates hypoxia-induced apoptotic cell death. In this paper, we unveil that the miR-210-induced inhibition of the serine/threonine kinase Glycogen Synthase Kinase 3 beta (GSK3β) in AC-16 cardiomyocytes subjected to hypoxia stress underlies the salutary protective response of miR-210 in mitigating the hypoxia-induced apoptotic cell death. Using transient overexpression vectors to augment miR-210 expression concomitant with the ectopic expression of the <i>constitutive active</i> GSK3β S9A mutant (<i>ca</i>-GSK3β S9A), we exhaustively performed biochemical and molecular assays to determine the status of the hypoxia-induced intrinsic apoptosis cascade. Caspase-3 activity analysis coupled with DNA fragmentation assays cogently demonstrate that the inhibition of GSK3β kinase activity underlies the miR-210-induced attenuation in the hypoxia-driven apoptotic cell death. Further elucidation and delineation of the upstream cellular events unveiled an indispensable role of the inhibition of GSK3β kinase activity in mediating the miR-210-induced mitigation of the hypoxia-driven BAX and BAK insertion into the <i>outer mitochondria membrane</i> (<i>OMM</i>) and the ensuing <i>Cytochrome C</i> release into the cytosol. Our study is the first to unveil that the inhibition of GSK3β kinase activity is indispensable in mediating the miR-210-orchestrated protective cellular response to hypoxia-induced apoptotic cell death.https://www.mdpi.com/1422-0067/23/16/9375miR-210hypoxiaAC-16 cardiomyocytesapoptosisGSK3βBAX |
| spellingShingle | Gurdeep Marwarha Øystein Røsand Katrine Hordnes Slagsvold Morten Andre Høydal GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia International Journal of Molecular Sciences miR-210 hypoxia AC-16 cardiomyocytes apoptosis GSK3β BAX |
| title | GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia |
| title_full | GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia |
| title_fullStr | GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia |
| title_full_unstemmed | GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia |
| title_short | GSK3β Inhibition Is the Molecular Pivot That Underlies the Mir-210-Induced Attenuation of Intrinsic Apoptosis Cascade during Hypoxia |
| title_sort | gsk3β inhibition is the molecular pivot that underlies the mir 210 induced attenuation of intrinsic apoptosis cascade during hypoxia |
| topic | miR-210 hypoxia AC-16 cardiomyocytes apoptosis GSK3β BAX |
| url | https://www.mdpi.com/1422-0067/23/16/9375 |
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