Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells
Neuroglobin (NGB) is an O<sub>2</sub>-binding globin mainly expressed in the central and peripheral nervous systems and cerebrospinal fluid. Previously, it was demonstrated that NGB overexpression protects cells from hypoxia-induced death. To investigate processes promoted by NGB overexp...
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MDPI AG
2021-12-01
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| Online Access: | https://www.mdpi.com/2073-4409/10/12/3394 |
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| author | Valeria Manganelli Illari Salvatori Michele Costanzo Antonella Capozzi Daniela Caissutti Marianna Caterino Cristiana Valle Alberto Ferri Maurizio Sorice Margherita Ruoppolo Tina Garofalo Roberta Misasi |
| author_facet | Valeria Manganelli Illari Salvatori Michele Costanzo Antonella Capozzi Daniela Caissutti Marianna Caterino Cristiana Valle Alberto Ferri Maurizio Sorice Margherita Ruoppolo Tina Garofalo Roberta Misasi |
| author_sort | Valeria Manganelli |
| collection | DOAJ |
| description | Neuroglobin (NGB) is an O<sub>2</sub>-binding globin mainly expressed in the central and peripheral nervous systems and cerebrospinal fluid. Previously, it was demonstrated that NGB overexpression protects cells from hypoxia-induced death. To investigate processes promoted by NGB overexpression, we used a cellular model of neuroblastoma stably overexpressing an NGB-FLAG construct. We used a proteomic approach to identify the specific profile following NGB overexpression. To evaluate the role of NGB overexpression in increasing energetic metabolism, we measured oxygen consumption rate (OCR) and the extracellular acidification rate through Seahorse XF technology. The effect on autophagy induction was evaluated by analyzing SQSTM1/p62 and LC3-II expression. Proteomic analysis revealed several differentially regulated proteins, involved in oxidative phosphorylation and integral mitochondrial proteins linked to energy metabolism. The analysis of mitochondrial metabolism demonstrated that NGB overexpression increases mitochondrial ATP production. Indeed, NGB overexpression enhances bioenergetic metabolism, increasing OCR and oxygen consumption. Analysis of autophagy induction revealed an increase of LC3-II together with a significant decrease of SQSTM1/p62, and NGB-LC3-II association during autophagosome formation. These results highlight the active participation of NGB in several cellular processes that can be upregulated in response to NGB overexpression, playing a role in the adaptive response to stress in neuroblastoma cells. |
| first_indexed | 2024-03-10T04:25:57Z |
| format | Article |
| id | doaj.art-95fb4d7e4420408aae6bef968400b480 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T04:25:57Z |
| publishDate | 2021-12-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-95fb4d7e4420408aae6bef968400b4802023-11-23T07:37:15ZengMDPI AGCells2073-44092021-12-011012339410.3390/cells10123394Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y CellsValeria Manganelli0Illari Salvatori1Michele Costanzo2Antonella Capozzi3Daniela Caissutti4Marianna Caterino5Cristiana Valle6Alberto Ferri7Maurizio Sorice8Margherita Ruoppolo9Tina Garofalo10Roberta Misasi11Department of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyFondazione Santa Lucia IRCCS, c/o CERC, 00143 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, ItalyDepartment of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, ItalyFondazione Santa Lucia IRCCS, c/o CERC, 00143 Rome, ItalyFondazione Santa Lucia IRCCS, c/o CERC, 00143 Rome, ItalyDepartment of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyDepartment of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131 Naples, ItalyDepartment of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyDepartment of Experimental Medicine, Sapienza University of Rome, 00161 Rome, ItalyNeuroglobin (NGB) is an O<sub>2</sub>-binding globin mainly expressed in the central and peripheral nervous systems and cerebrospinal fluid. Previously, it was demonstrated that NGB overexpression protects cells from hypoxia-induced death. To investigate processes promoted by NGB overexpression, we used a cellular model of neuroblastoma stably overexpressing an NGB-FLAG construct. We used a proteomic approach to identify the specific profile following NGB overexpression. To evaluate the role of NGB overexpression in increasing energetic metabolism, we measured oxygen consumption rate (OCR) and the extracellular acidification rate through Seahorse XF technology. The effect on autophagy induction was evaluated by analyzing SQSTM1/p62 and LC3-II expression. Proteomic analysis revealed several differentially regulated proteins, involved in oxidative phosphorylation and integral mitochondrial proteins linked to energy metabolism. The analysis of mitochondrial metabolism demonstrated that NGB overexpression increases mitochondrial ATP production. Indeed, NGB overexpression enhances bioenergetic metabolism, increasing OCR and oxygen consumption. Analysis of autophagy induction revealed an increase of LC3-II together with a significant decrease of SQSTM1/p62, and NGB-LC3-II association during autophagosome formation. These results highlight the active participation of NGB in several cellular processes that can be upregulated in response to NGB overexpression, playing a role in the adaptive response to stress in neuroblastoma cells.https://www.mdpi.com/2073-4409/10/12/3394neuroglobinmitochondriaautophagyenergy metabolismSH-SY5Y neuroblastoma cellslabel-free proteomics |
| spellingShingle | Valeria Manganelli Illari Salvatori Michele Costanzo Antonella Capozzi Daniela Caissutti Marianna Caterino Cristiana Valle Alberto Ferri Maurizio Sorice Margherita Ruoppolo Tina Garofalo Roberta Misasi Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells Cells neuroglobin mitochondria autophagy energy metabolism SH-SY5Y neuroblastoma cells label-free proteomics |
| title | Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells |
| title_full | Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells |
| title_fullStr | Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells |
| title_full_unstemmed | Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells |
| title_short | Overexpression of Neuroglobin Promotes Energy Metabolism and Autophagy Induction in Human Neuroblastoma SH-SY5Y Cells |
| title_sort | overexpression of neuroglobin promotes energy metabolism and autophagy induction in human neuroblastoma sh sy5y cells |
| topic | neuroglobin mitochondria autophagy energy metabolism SH-SY5Y neuroblastoma cells label-free proteomics |
| url | https://www.mdpi.com/2073-4409/10/12/3394 |
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