Neuroglobin (NGB) is an O2-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 overexpres-sion, 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 expres-sion. 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.

Overexpression of neuroglobin promotes energy metabolism and autophagy induction in human neuroblastoma SH-SY5Y cells / Manganelli, V.; Salvatori, I.; Costanzo, M.; Capozzi, A.; Caissutti, D.; Caterino, M.; Valle, C.; Ferri, A.; Sorice, M.; Ruoppolo, M.; Garofalo, T.; Misasi, R.. - In: CELLS. - ISSN 2073-4409. - 10:12(2021), pp. 1-18. [10.3390/cells10123394]

Overexpression of neuroglobin promotes energy metabolism and autophagy induction in human neuroblastoma SH-SY5Y cells

Manganelli V.
Co-primo
Investigation
;
Salvatori I.
Co-primo
Investigation
;
Capozzi A.
Validation
;
Caissutti D.
Validation
;
Sorice M.
Writing – Original Draft Preparation
;
Garofalo T.
Writing – Original Draft Preparation
;
Misasi R.
Ultimo
Writing – Original Draft Preparation
2021

Abstract

Neuroglobin (NGB) is an O2-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 overexpres-sion, 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 expres-sion. 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.
2021
autophagy; energy metabolism; label-free proteomics; mitochondria; neuroglobin; sh-sy5y neuroblastoma cells; adenosine triphosphate; autophagy; cell line; tumor; energy metabolism; gene expression regulation, neoplastic; humans; microtubule-associated proteins; mitochondria; neuroblastoma; neuroglobin; oxygen consumption; proteome; sequestosome-1 protein
01 Pubblicazione su rivista::01a Articolo in rivista
Overexpression of neuroglobin promotes energy metabolism and autophagy induction in human neuroblastoma SH-SY5Y cells / Manganelli, V.; Salvatori, I.; Costanzo, M.; Capozzi, A.; Caissutti, D.; Caterino, M.; Valle, C.; Ferri, A.; Sorice, M.; Ruoppolo, M.; Garofalo, T.; Misasi, R.. - In: CELLS. - ISSN 2073-4409. - 10:12(2021), pp. 1-18. [10.3390/cells10123394]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/1614203
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