89 results on '"Cunha-Oliveira T"'
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2. Role of Mitochondria on the Neurological Effects of Cocaine
- Author
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Pereira, S.P., primary and Cunha-Oliveira, T., additional
- Published
- 2017
- Full Text
- View/download PDF
3. List of Contributors
- Author
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Alves, C.J., primary, Anglard, P., additional, Anier, K., additional, Armstrong, R.A., additional, Bachtell, R.K., additional, Bakshi, K., additional, Barbanti, P., additional, Barrio, P., additional, Batalla, A., additional, Becker, J.B., additional, Bekker, A., additional, Benkelfat, C., additional, Bergman, J., additional, Bhattacharya, P., additional, Bisagno, Veronica, additional, Brimijoin, S., additional, Brown, Z.J., additional, Buffalari, D., additional, Bühler, K.-M., additional, Caffino, L., additional, Cafforio, G., additional, Camarini, R., additional, Carvalho, V.M., additional, Cepko, L.C.S., additional, Chen, C.-C., additional, Chu, X.-P., additional, Corbit, L.H., additional, Crofton, E.J., additional, Crunelle, C.L., additional, Cunha, P.J., additional, Cunha-Oliveira, T., additional, Currie, P.J., additional, D’Ascenzo, M., additional, Dieckmann, L.H.J., additional, von Diemen, L., additional, Dumont, E.C., additional, Eagle, A.L., additional, Eipper, B.A., additional, Eipper-Mains, J.E., additional, Engeln, M., additional, Erb, S., additional, Farré, A., additional, Farré, M., additional, Felts, A.S., additional, Fofi, L., additional, Foster, J.D., additional, Fox, H.C., additional, Frankfurt, M., additional, Freissmuth, M., additional, Fuchs, R.A., additional, Fumagalli, F., additional, Gajewski, P.A., additional, Galaj, E., additional, Galduróz, J.C.F., additional, Garling, E.E., additional, Gentile, T.A., additional, Giannotti, G., additional, Girault, J.-A., additional, Glass, J.D., additional, Goncalves, P.D., additional, González-Duarte, A., additional, Gonzalez-Nunez, V., additional, Gould, R.W., additional, Grassi, C., additional, Green, T.A., additional, Green-Sadan, T., additional, Gu, H.H., additional, Guan, Xiaowei, additional, Halbout, B., additional, Han, D.D., additional, Henry, L.K., additional, Pérez de Heredia, J.L., additional, Higginbotham, J.A., additional, Hofmaier, T., additional, Holy, M., additional, Hsu, K.-S., additional, Huang, C.-C., additional, James, J., additional, Jones, A.W., additional, Jones, C.K., additional, Kalda, A., additional, Kearns, D.N., additional, Kerver, H.N., additional, Kessler, F., additional, Kohut, S.J., additional, Krnjević, K., additional, Kucab, P., additional, Kudlacek, O., additional, Kupferschmidt, D.A., additional, Kuzhikandathil, E.V., additional, Lee, M.R., additional, Leggio, L., additional, Lever, J.R., additional, Lever, S.Z., additional, Leyton, M., additional, Li, J.-X., additional, Lima, D.R., additional, Lobo, M.K., additional, López-Moreno, J.A., additional, López-Pelayo, H., additional, Lovejoy, D.A., additional, Luf, A., additional, Lugon, M.D.M.V., additional, Lyons, C.E., additional, Magalhães, A., additional, Magalhães, P.V.S., additional, Mainardi, M., additional, Mains, R.E., additional, Mantsch, J.R., additional, Marcourakis, T., additional, Marhe, R., additional, Matthys, F., additional, El Mestikawy, S., additional, Milivojevic, V., additional, Miller, D.K., additional, Min, M.O., additional, Minnes, S., additional, Mitchell, M.R., additional, Monteiro, P.R., additional, Murthy, V., additional, Muschamp, J.W., additional, Nagy, C., additional, Nakamura-Palacios, E.M., additional, Narvaez, J.C.M., additional, Normandeau, C.P., additional, Ornell, F., additional, Orsini, C.A., additional, Ostlund, S.B., additional, Otkins, J., additional, Patel, V.B., additional, Pelição, F.S., additional, Pereira, S.P., additional, Peres, M.D., additional, Potenza, M.N., additional, Preedy, V.R., additional, Prosser, R.A., additional, Quednow, B.B., additional, Rajendram, R., additional, Ramos, A.C., additional, Ranaldi, R., additional, Rangel-Barajas, C., additional, Rebec, G.V., additional, Robison, A.J., additional, Rodríguez, R.E., additional, Rohn, M.C.H., additional, Roth-Deri, I., additional, Scala, S.G., additional, Scavone, C., additional, Schellekens, A., additional, Scherer, J., additional, Schmid, R., additional, Scurlock, R.D., additional, Setlow, B., additional, Simmons, S.J., additional, Singer, L.T., additional, Sinha, R., additional, Sitte, H.H., additional, Smart, K., additional, Stockner, T., additional, Summavielle, T., additional, Szumlinski, K.K., additional, Tanda, G., additional, Torrens, M., additional, Tunstall, B.J., additional, Urbano, F.J., additional, Vaughan, R.A., additional, Verhaeghe, M., additional, Vonmoos, M., additional, Wagner, J.J., additional, Wang, Y., additional, Xi, Z.-X., additional, Xu, Y., additional, Yadid, G., additional, Ye, J.-H., additional, Yoon, S., additional, Zallar, L.J., additional, Zhan, Chang-Guo, additional, Zhang, H.-Y., additional, Zhang, Y., additional, Zheng, Fang, additional, Zuo, W., additional, and Zwiller, J., additional
- Published
- 2017
- Full Text
- View/download PDF
4. Metabolic remodeling of human skin fibroblasts as model for studying mitochondrial diseases and drug-induced toxicity: 5.40
- Author
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Pereira, S. P., Serafim, T. L., Cunha-Oliveira, T., Deus, C. M., Starostina, I. G., Ivanova, V. V., Rizvanov, A. A., and Oliveira, P. J.
- Published
- 2016
5. Decreased global DNA methylation levels and gene expression patterns related metabolic end epigenetic remodeling enzymes are observed after Doxorubicin sub-chronic treatment: 5.38
- Author
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Ferreira, A. M., Simöes, R. F., Vilaça, A., Carvalho, F. S., Garcia, R., Burgeiro, A., Cunha-Oliveira, T., and Oliveira, P. J.
- Published
- 2016
6. Causes and consequences of mitochondrial dysfunction induced by drugs of abuse: 5.14
- Author
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Cunha-Oliveira, T.
- Published
- 2016
7. Maternal nutrition and fetal mitochondrial fitness: 3.46
- Author
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Pereira, S. P., Tavares, L. C., Duarte, A. I., Cunha-Oliveira, T., Santos, M. S., Baldeiras, I., Clarke, G., Moreno, A. J., Cox, L. A., Nathanielsz, P. W., Nijland, M. J., and Oliveira, P. J.
- Published
- 2016
8. Doxorubicin treatment induces persistent alterations in cardiomyoblast mitochondrial gene expression: 3.20
- Author
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Ferreira, L. L., Cunha-Oliveira, T., and Oliveira, P. J.
- Published
- 2015
9. Mitochondrial complex I dysfunction induced by cocaine and cocaine plus morphine in brain and liver mitochondria: 2.37
- Author
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Cunha-Oliveira, T., Silva, L., Silva, A. M., Moreno, A. J., Oliveira, C. R., and Santos, M. S.
- Published
- 2013
10. Role of histone deacetylase inhibitors on pyruvate dehydrogenase dysfunction in Huntingtonʼs disease striatal cells: 2.15
- Author
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Rodrigues, J., Cunha-Oliveira, T., Ribeiro, M., and Rego, A. C.
- Published
- 2013
11. Reactivation of Dihydroorotate Dehydrogenase-Driven Pyrimidine Biosynthesis Restores Tumor Growth of Respiration-Deficient Cancer Cells
- Author
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Bajzikova, M., Kovarova, J., Coelho, A.R., Boukalova, S., Oh, S., Rohlenova, K., Svec, D., Hubackova, S., Endaya, B., Judasova, K., Bezawork-Geleta, A., Kluckova, K., Chatre, L., Zobalova, R., Novakova, A., Vanova, K., Ezrova, Z., Maghzal, G.J., Novais, S. Magalhaes, Olsinova, M., Krobova, L., An, Y.J., Davidova, E., Nahacka, Z., Sobol, M., Cunha-Oliveira, T., Sandoval-Acuna, C., Strnad, H., Zhang, T., Huynh, T., Serafim, T.L., Hozak, P., Sardao, V.A., Koopman, W.J.H., Ricchetti, M., Oliveira, P.J., Kolar, F., Kubista, M., Truksa, J., Dvorakova-Hortova, K., Pacak, K., Gurlich, R., Stocker, R., Zhou, Y, Berridge, M.V., Park, S., Dong, L., Rohlena, J., Neuzil, J., Bajzikova, M., Kovarova, J., Coelho, A.R., Boukalova, S., Oh, S., Rohlenova, K., Svec, D., Hubackova, S., Endaya, B., Judasova, K., Bezawork-Geleta, A., Kluckova, K., Chatre, L., Zobalova, R., Novakova, A., Vanova, K., Ezrova, Z., Maghzal, G.J., Novais, S. Magalhaes, Olsinova, M., Krobova, L., An, Y.J., Davidova, E., Nahacka, Z., Sobol, M., Cunha-Oliveira, T., Sandoval-Acuna, C., Strnad, H., Zhang, T., Huynh, T., Serafim, T.L., Hozak, P., Sardao, V.A., Koopman, W.J.H., Ricchetti, M., Oliveira, P.J., Kolar, F., Kubista, M., Truksa, J., Dvorakova-Hortova, K., Pacak, K., Gurlich, R., Stocker, R., Zhou, Y, Berridge, M.V., Park, S., Dong, L., Rohlena, J., and Neuzil, J.
- Abstract
Contains fulltext : 202265.pdf (publisher's version ) (Closed access), Cancer cells without mitochondrial DNA (mtDNA) do not form tumors unless they reconstitute oxidative phosphorylation (OXPHOS) by mitochondria acquired from host stroma. To understand why functional respiration is crucial for tumorigenesis, we used time-resolved analysis of tumor formation by mtDNA-depleted cells and genetic manipulations of OXPHOS. We show that pyrimidine biosynthesis dependent on respiration-linked dihydroorotate dehydrogenase (DHODH) is required to overcome cell-cycle arrest, while mitochondrial ATP generation is dispensable for tumorigenesis. Latent DHODH in mtDNA-deficient cells is fully activated with restoration of complex III/IV activity and coenzyme Q redox-cycling after mitochondrial transfer, or by introduction of an alternative oxidase. Further, deletion of DHODH interferes with tumor formation in cells with fully functional OXPHOS, while disruption of mitochondrial ATP synthase has little effect. Our results show that DHODH-driven pyrimidine biosynthesis is an essential pathway linking respiration to tumorigenesis, pointing to inhibitors of DHODH as potential anti-cancer agents.
- Published
- 2019
12. Chapter 21 - Role of Mitochondria on the Neurological Effects of Cocaine
- Author
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Pereira, S.P. and Cunha-Oliveira, T.
- Published
- 2017
- Full Text
- View/download PDF
13. Role of Mitochondria on the Neurological Effects of Cocaine
- Author
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Pereira, S.P. and Cunha-Oliveira, T.
- Published
- 2017
14. Berberine-induced cardioprotection and Sirt3 modulation in doxorubicin-treated H9c2 cardiomyoblasts
- Author
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Coelho A., Martins T., Couto R., Deus C., Pereira C., Simões R., Rizvanov A., Silva F., Cunha-Oliveira T., Oliveira P., and Serafim T.
- Subjects
carbohydrates (lipids) ,Berberine ,Doxorubicin ,organic chemicals ,Sirtuin 3 ,technology, industry, and agriculture ,polycyclic compounds ,macromolecular substances ,Cardiotoxicity - Abstract
© 2017 Elsevier B.V. Doxorubicin (DOX) is one of the most widely used anti-neoplastic agents. However, treatment with DOX is associated with cumulative cardiotoxicity inducing progressive cardiomyocyte death. Sirtuin 3 (Sirt3), a mitochondrial deacetylase, regulates the activity of proteins involved in apoptosis, autophagy and metabolism. Our hypothesis is that pharmacological modulation by berberine (BER) pre-conditioning of Sirt3 protein levels decreases DOX-induced cardiotoxicity. Our results showed that DOX induces cell death in all experimental groups. Increase in Sirt3 content by transfection-mediated overexpression decreased DOX cytotoxicity, mostly by maintaining mitochondrial network integrity and reducing oxidative stress. p53 was upregulated by DOX, and appeared to be a direct target of Sirt3, suggesting that Sirt3-mediated protection against cell death could be related to this protein. BER pre-treatment increased Sirt3 and Sirt1 protein levels in the presence of DOX and inhibited DOX-induced caspase 9 and 3-like activation. Moreover, BER modulated autophagy in DOX-treated H9c2 cardiomyoblasts. Interestingly, mitochondrial biogenesis markers were upregulated in in BER/DOX-treated cells. Sirt3 over-expression contributes to decrease DOX cytotoxicity on H9c2 cardiomyoblasts, while BER can be used as a modulator of Sirtuin function and cell quality control pathways to decrease DOX toxicity.
- Published
- 2017
15. β-adrenergic over-stimulation and cardio-myocyte apoptosis: Two receptors, one organelle, two fates?
- Author
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Branco A., Moreira A., Cunha-Oliveira T., Couto R., Sardão V., Rizvanov A., Palotás A., and Oliveira P.
- Subjects
Oxidative stress ,Apoptosis ,Calcium ,Cardio-myocyte ,β-adrenergic receptors ,Mitochondria - Abstract
© 2014 Bentham Science Publishers. Neuro-hormonal regulation of cardiac function via cathecol-amines results in increased heart rate and contractility. A persistent adrenergic tone, however, is an insult to the heart, affecting its regular homeostasis, altering morphology and gene expression patterns, as well as inducing apoptosis of cardio-myocytes. At the same time as being the main oxygen consumers, mitochondria are also key to the energy production required for the heart to maintain its vital functions and to integrate a series of signaling pathways that define the life and death of the cell. As β-adrenergic receptors (β-AR) orchestrate multiple biochemical events that can either trigger or inhibit cell death, mitochondria can act as a referee in the entire process. In fact, β-AR subtypes β1 and β2 activate various down-stream pathways which differently modulate intracellular calcium levels and production of mitochondrial reactive oxygen species (ROS). The delicate balance between an adaptive (cardio-protective) response resulting in increased contractility and activation of survival pathways, vs. cell death caused by calcium and ROS-induced mitochondrial disruption, along with evidence of their clinical and potential therapeutic translations, are reviewed in this communication.
- Published
- 2014
16. Adaptation to oxidative stress in the chronic effects of cocaine and amphetamine
- Author
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Cunha-Oliveira, T., Rego, A. C., Macedo, T., and Oliveira, C. R.
- Published
- 2007
17. Differential cytotoxic responses of PC12 cells chronically exposed to psychostimulants or to hydrogen peroxide
- Author
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Cunha-Oliveira, T., Rego, A. C., Morgadinho, M. T., Sarmento, A., Macedo, T., and Oliveira, C. R.
- Published
- 2005
18. Molecular interaction between cocaine and opioids. Implications on Speedball's toxicity
- Author
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Cunha-Oliveira, T., primary, Silva, L., additional, Silva, A.M., additional, Santos, M.S., additional, Garrido, J., additional, Borges, F., additional, Rego, A.C., additional, and Oliveira, C.R., additional
- Published
- 2011
- Full Text
- View/download PDF
19. Expression of NR1/NR2B N-Methyl-D-Aspartate Receptors Enhances Heroin Toxicity in HEK293 Cells
- Author
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DOMINGUES, A., primary, CUNHA OLIVEIRA, T., additional, LACO, M. L. N, additional, MACEDO, T. R. A, additional, OLIVEIRA, C. R, additional, and REGO, A. C., additional
- Published
- 2006
- Full Text
- View/download PDF
20. ß-adrenergic over-stimulation and cardio-myocyte apoptosis: Two receptors, one organelle, two fates?
- Author
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Branco, A. F., Moreira, A. C., Cunha-Oliveira, T., Couto, R., Vilma Sardao, Rizvanov, A. A., Palotás, A., and Oliveira, P. J.
21. Metabolic remodeling of human skin fibroblasts as model for studying mitochondrial diseases and drug-induced toxicity
- Author
-
Pereira, S. P., Serafim, T. L., Cunha-Oliveira, T., Deus, C. M., Starostina, I. G., Ivanova, V. V., Albert Rizvanov, and Oliveira, P. J.
22. β-adrenergic over-stimulation and cardio-myocyte apoptosis: two receptors, one organelle, two fates?
- Author
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Ana Filipa Branco, Ac, Moreira, Cunha-Oliveira T, Couto R, Va, Sardao, Aa, Rizvanov, Palotas A, and Pj, Oliveira
23. β-adrenergic over-stimulation and cardio-myocyte apoptosis: Two receptors, one organelle, two fates?
- Author
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Branco A., Moreira A., Cunha-Oliveira T., Couto R., Sardão V., Rizvanov A., Palotás A., Oliveira P., Branco A., Moreira A., Cunha-Oliveira T., Couto R., Sardão V., Rizvanov A., Palotás A., and Oliveira P.
- Abstract
© 2014 Bentham Science Publishers. Neuro-hormonal regulation of cardiac function via cathecol-amines results in increased heart rate and contractility. A persistent adrenergic tone, however, is an insult to the heart, affecting its regular homeostasis, altering morphology and gene expression patterns, as well as inducing apoptosis of cardio-myocytes. At the same time as being the main oxygen consumers, mitochondria are also key to the energy production required for the heart to maintain its vital functions and to integrate a series of signaling pathways that define the life and death of the cell. As β-adrenergic receptors (β-AR) orchestrate multiple biochemical events that can either trigger or inhibit cell death, mitochondria can act as a referee in the entire process. In fact, β-AR subtypes β1 and β2 activate various down-stream pathways which differently modulate intracellular calcium levels and production of mitochondrial reactive oxygen species (ROS). The delicate balance between an adaptive (cardio-protective) response resulting in increased contractility and activation of survival pathways, vs. cell death caused by calcium and ROS-induced mitochondrial disruption, along with evidence of their clinical and potential therapeutic translations, are reviewed in this communication.
24. β-adrenergic over-stimulation and cardio-myocyte apoptosis: Two receptors, one organelle, two fates?
- Author
-
Branco A., Moreira A., Cunha-Oliveira T., Couto R., Sardão V., Rizvanov A., Palotás A., Oliveira P., Branco A., Moreira A., Cunha-Oliveira T., Couto R., Sardão V., Rizvanov A., Palotás A., and Oliveira P.
- Abstract
© 2014 Bentham Science Publishers. Neuro-hormonal regulation of cardiac function via cathecol-amines results in increased heart rate and contractility. A persistent adrenergic tone, however, is an insult to the heart, affecting its regular homeostasis, altering morphology and gene expression patterns, as well as inducing apoptosis of cardio-myocytes. At the same time as being the main oxygen consumers, mitochondria are also key to the energy production required for the heart to maintain its vital functions and to integrate a series of signaling pathways that define the life and death of the cell. As β-adrenergic receptors (β-AR) orchestrate multiple biochemical events that can either trigger or inhibit cell death, mitochondria can act as a referee in the entire process. In fact, β-AR subtypes β1 and β2 activate various down-stream pathways which differently modulate intracellular calcium levels and production of mitochondrial reactive oxygen species (ROS). The delicate balance between an adaptive (cardio-protective) response resulting in increased contractility and activation of survival pathways, vs. cell death caused by calcium and ROS-induced mitochondrial disruption, along with evidence of their clinical and potential therapeutic translations, are reviewed in this communication.
25. Berberine-induced cardioprotection and Sirt3 modulation in doxorubicin-treated H9c2 cardiomyoblasts
- Author
-
Coelho A., Martins T., Couto R., Deus C., Pereira C., Simões R., Rizvanov A., Silva F., Cunha-Oliveira T., Oliveira P., Serafim T., Coelho A., Martins T., Couto R., Deus C., Pereira C., Simões R., Rizvanov A., Silva F., Cunha-Oliveira T., Oliveira P., and Serafim T.
- Abstract
© 2017 Elsevier B.V. Doxorubicin (DOX) is one of the most widely used anti-neoplastic agents. However, treatment with DOX is associated with cumulative cardiotoxicity inducing progressive cardiomyocyte death. Sirtuin 3 (Sirt3), a mitochondrial deacetylase, regulates the activity of proteins involved in apoptosis, autophagy and metabolism. Our hypothesis is that pharmacological modulation by berberine (BER) pre-conditioning of Sirt3 protein levels decreases DOX-induced cardiotoxicity. Our results showed that DOX induces cell death in all experimental groups. Increase in Sirt3 content by transfection-mediated overexpression decreased DOX cytotoxicity, mostly by maintaining mitochondrial network integrity and reducing oxidative stress. p53 was upregulated by DOX, and appeared to be a direct target of Sirt3, suggesting that Sirt3-mediated protection against cell death could be related to this protein. BER pre-treatment increased Sirt3 and Sirt1 protein levels in the presence of DOX and inhibited DOX-induced caspase 9 and 3-like activation. Moreover, BER modulated autophagy in DOX-treated H9c2 cardiomyoblasts. Interestingly, mitochondrial biogenesis markers were upregulated in in BER/DOX-treated cells. Sirt3 over-expression contributes to decrease DOX cytotoxicity on H9c2 cardiomyoblasts, while BER can be used as a modulator of Sirtuin function and cell quality control pathways to decrease DOX toxicity.
26. Berberine-induced cardioprotection and Sirt3 modulation in doxorubicin-treated H9c2 cardiomyoblasts
- Author
-
Coelho A., Martins T., Couto R., Deus C., Pereira C., Simões R., Rizvanov A., Silva F., Cunha-Oliveira T., Oliveira P., Serafim T., Coelho A., Martins T., Couto R., Deus C., Pereira C., Simões R., Rizvanov A., Silva F., Cunha-Oliveira T., Oliveira P., and Serafim T.
- Abstract
© 2017 Elsevier B.V. Doxorubicin (DOX) is one of the most widely used anti-neoplastic agents. However, treatment with DOX is associated with cumulative cardiotoxicity inducing progressive cardiomyocyte death. Sirtuin 3 (Sirt3), a mitochondrial deacetylase, regulates the activity of proteins involved in apoptosis, autophagy and metabolism. Our hypothesis is that pharmacological modulation by berberine (BER) pre-conditioning of Sirt3 protein levels decreases DOX-induced cardiotoxicity. Our results showed that DOX induces cell death in all experimental groups. Increase in Sirt3 content by transfection-mediated overexpression decreased DOX cytotoxicity, mostly by maintaining mitochondrial network integrity and reducing oxidative stress. p53 was upregulated by DOX, and appeared to be a direct target of Sirt3, suggesting that Sirt3-mediated protection against cell death could be related to this protein. BER pre-treatment increased Sirt3 and Sirt1 protein levels in the presence of DOX and inhibited DOX-induced caspase 9 and 3-like activation. Moreover, BER modulated autophagy in DOX-treated H9c2 cardiomyoblasts. Interestingly, mitochondrial biogenesis markers were upregulated in in BER/DOX-treated cells. Sirt3 over-expression contributes to decrease DOX cytotoxicity on H9c2 cardiomyoblasts, while BER can be used as a modulator of Sirtuin function and cell quality control pathways to decrease DOX toxicity.
27. Characterization of the Mitochondria Function and Metabolism in Skin Fibroblasts Using the Biolog MitoPlate S-1.
- Author
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de Lemos AC, Teixeira J, and Cunha-Oliveira T
- Subjects
- Humans, Oxidation-Reduction, NAD metabolism, Electron Transport, Cells, Cultured, Fibroblasts metabolism, Mitochondria metabolism, Skin metabolism, Skin cytology
- Abstract
S-1 MitoPlates™ from Biolog enable the characterization of mitochondria's function in live cells by measuring the rates of electron flow into and through the electron transport chain from different NADH or FADH
2 producing metabolic substrates. This technology uses 96-well microplates pre-coated with triplicate repeats of a set of 31 substrates. Those 31 metabolic substrates have different routes of entry into the mitochondria, use different transporters, and are also oxidated by different dehydrogenases, producing reducing equivalents in the form of NADH or FADH2 . The electrons produced upon oxidation of NADH or FADH2 at complex I or II, respectively, then travel to cytochrome c, where a tetrazolium redox dye (MC) can act as terminal acceptor, turning purple and absorbing at 590 nm. This mechanism allows the evaluation of cellular substrate preference by following the kinetics of MC reduction in the presence of selected substrates.In this chapter, we describe the step-by-step protocol to prepare an experiment using MitoPlate S-1 array and the OmniLog instrument to assess the metabolism of human dermal fibroblasts. We also give detailed information on how to analyze the raw data generated by the Biolog Data Analysis software to extract meaningful information and produce useful data visualizations, using reproducible methods based on a single structured dataset., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2025
- Full Text
- View/download PDF
28. Creating Physiological Cell Environments In Vitro: Adjusting Cell Culture Media Composition and Oxygen Levels to Investigate Mitochondrial Function and Cancer Metabolism.
- Author
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Pinho SA, Gardner GL, Alva R, Stuart JA, and Cunha-Oliveira T
- Subjects
- Humans, Energy Metabolism drug effects, Glucose metabolism, Mitochondria metabolism, Mitochondria drug effects, Oxygen metabolism, Culture Media chemistry, Fibroblasts metabolism, Fibroblasts drug effects, Cell Culture Techniques methods, Tumor Microenvironment drug effects, Neoplasms metabolism, Neoplasms drug therapy, Neoplasms pathology
- Abstract
In vitro and ex vivo studies are crucial for mitochondrial research, offering valuable insights into cellular mechanisms and aiding in diagnostic and therapeutic strategies. Accurate in vitro models rely on adequate cell culture conditions, such as the composition of culture media and oxygenation levels. These conditions can influence energy metabolism and mitochondrial activities, thus impacting studies involving mitochondrial components, such as the effectiveness of anticancer drugs. This chapter focuses on practical guidance for creating setups that replicate in vivo microenvironments, capturing the original metabolic context of cells. We explore protocols to better mimic the physiological cell environment, promote cellular reconfiguration, and prime cells according to the modeled context. The first part is dedicated to the use of human dermal fibroblasts, which are a promising model for pre-clinical mitochondrial research due to their adaptability and relevance to human mitochondrial physiology. We present an optimized protocol for gradually adjusting extracellular glucose levels, which demonstrated significant mitochondrial, metabolic, and redox remodeling in normal adult dermal fibroblasts. The second part is dedicated to replication of tumor microenvironments, which are relevant for studies targeting cellular energy metabolism to inhibit tumor growth. Currently available physiological media can mimic blood plasma metabolome but not the specific tumor microenvironment. To address this, we describe optimized media formulation and oxygenation protocols, which can simulate the tumor microenvironment in cell culture experiments. Replicating in vivo microenvironments in in vitro and ex vivo studies can enhance our understanding of cellular processes, facilitate drug development, and advance personalized therapeutics in mitochondrial medicine., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2025
- Full Text
- View/download PDF
29. Heterogeneous redox responses in NHDF cells primed to enhance mitochondrial bioenergetics.
- Author
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Pinho SA, Oliveira PJ, and Cunha-Oliveira T
- Subjects
- Humans, Reactive Oxygen Species metabolism, Oxidative Phosphorylation drug effects, Cells, Cultured, Cell Cycle drug effects, Oxidation-Reduction drug effects, Fibroblasts metabolism, Fibroblasts drug effects, Oxidative Stress drug effects, Mitochondria metabolism, Mitochondria drug effects, Energy Metabolism drug effects, Hydrogen Peroxide pharmacology, Hydrogen Peroxide metabolism, tert-Butylhydroperoxide pharmacology
- Abstract
Aging and lifestyle-related diseases, such as cardiovascular diseases, diabetes, cancer, and neurodegenerative disorders, are major global health challenges. These conditions are often linked to redox imbalances, where cells fail to regulate reactive redox species (RRS), leading to oxidative stress and cellular damage. Although antioxidants are known to neutralize harmful RRS, their clinical efficacy remains inconsistent. One reason for this inconsistency is the inadequacy of current in vitro models to accurately mimic in vivo redox conditions. This study addresses the gap in understanding the heterogeneity of redox responses in cells by using metabolically primed human dermal fibroblasts (NHDF), a model relevant for precision mitochondrial medicine. We investigated how metabolic priming, which enhances mitochondrial bioenergetics, influences redox responses to oxidative stress induced by hydrogen peroxide (H
2 O2 ) and tert-butyl hydroperoxide (tBHP). Specifically, we explored the impact of cell population density and cell cycle distribution on redox dynamics. Our findings indicate that NHDF cells cultured in oxidative phosphorylation-promoting medium (OXm) exhibit significantly larger variability in oxidative stress responses. This variability suggests that enhanced mitochondrial bioenergetics necessitates a constant regulation of the cellular redox machinery, potentially leading to heterogeneous responses. Additionally, cells grown in OXm showed increased mitochondrial polarization and a lower percentage of cells in the G2/M phase, contributing to the observed heterogeneity. Key factors influencing this variability included cell population density at the time of oxidant exposure and fluctuations in cell cycle distribution. Our results highlight the necessity of employing multiple oxidants in metabolic priming models to achieve a comprehensive understanding of oxidative stress responses and redox regulation mechanisms. Furthermore, the study emphasizes the need to refine in vitro models to better reflect in vivo conditions, which is crucial for the development of effective redox-based therapeutic strategies., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Teresa Cunha-Oliveira reports financial support was provided by FCT - Foundation for Science and Technology. Paulo J. Oliveira reports a relationship with MitoTAG that includes: equity or stocks. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)- Published
- 2025
- Full Text
- View/download PDF
30. Best practices for data management and sharing in experimental biomedical research.
- Author
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Cunha-Oliveira T, Ioannidis JPA, and Oliveira PJ
- Subjects
- Humans, Animals, Biomedical Research standards, Biomedical Research methods, Information Dissemination methods, Data Management methods
- Abstract
Effective data management is crucial for scientific integrity and reproducibility, a cornerstone of scientific progress. Well-organized and well-documented data enable validation and building on results. Data management encompasses activities including organization, documentation, storage, sharing, and preservation. Robust data management establishes credibility, fostering trust within the scientific community and benefiting researchers' careers. In experimental biomedicine, comprehensive data management is vital due to the typically intricate protocols, extensive metadata, and large datasets. Low-throughput experiments, in particular, require careful management to address variations and errors in protocols and raw data quality. Transparent and accountable research practices rely on accurate documentation of procedures, data collection, and analysis methods. Proper data management ensures long-term preservation and accessibility of valuable datasets. Well-managed data can be revisited, contributing to cumulative knowledge and potential new discoveries. Publicly funded research has an added responsibility for transparency, resource allocation, and avoiding redundancy. Meeting funding agency expectations increasingly requires rigorous methodologies, adherence to standards, comprehensive documentation, and widespread sharing of data, code, and other auxiliary resources. This review provides critical insights into raw and processed data, metadata, high-throughput versus low-throughput datasets, a common language for documentation, experimental and reporting guidelines, efficient data management systems, sharing practices, and relevant repositories. We systematically present available resources and optimal practices for wide use by experimental biomedical researchers.
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- 2024
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31. Mitochondria: A Promising Convergent Target for the Treatment of Amyotrophic Lateral Sclerosis.
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Cunha-Oliveira T, Montezinho L, Simões RF, Carvalho M, Ferreiro E, and Silva FSG
- Subjects
- Humans, Mitochondria metabolism, Motor Neurons pathology, Apoptosis, Amyotrophic Lateral Sclerosis metabolism, Neurodegenerative Diseases metabolism
- Abstract
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease characterized by the progressive loss of motor neurons, for which current treatment options are limited. Recent studies have shed light on the role of mitochondria in ALS pathogenesis, making them an attractive therapeutic intervention target. This review contains a very comprehensive critical description of the involvement of mitochondria and mitochondria-mediated mechanisms in ALS. The review covers several key areas related to mitochondria in ALS, including impaired mitochondrial function, mitochondrial bioenergetics, reactive oxygen species, metabolic processes and energy metabolism, mitochondrial dynamics, turnover, autophagy and mitophagy, impaired mitochondrial transport, and apoptosis. This review also highlights preclinical and clinical studies that have investigated various mitochondria-targeted therapies for ALS treatment. These include strategies to improve mitochondrial function, such as the use of dichloroacetate, ketogenic and high-fat diets, acetyl-carnitine, and mitochondria-targeted antioxidants. Additionally, antiapoptotic agents, like the mPTP-targeting agents minocycline and rasagiline, are discussed. The paper aims to contribute to the identification of effective mitochondria-targeted therapies for ALS treatment by synthesizing the current understanding of the role of mitochondria in ALS pathogenesis and reviewing potential convergent therapeutic interventions. The complex interplay between mitochondria and the pathogenic mechanisms of ALS holds promise for the development of novel treatment strategies to combat this devastating disease.
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- 2024
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32. Characterizing Early Cardiac Metabolic Programming via 30% Maternal Nutrient Reduction during Fetal Development in a Non-Human Primate Model.
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Pereira SP, Diniz MS, Tavares LC, Cunha-Oliveira T, Li C, Cox LA, Nijland MJ, Nathanielsz PW, and Oliveira PJ
- Subjects
- Pregnancy, Humans, Animals, Male, Female, Fetus metabolism, Fetal Growth Retardation metabolism, Primates, Nutrients, Fetal Development, Cardiovascular Diseases metabolism
- Abstract
Intra-uterine growth restriction (IUGR) is a common cause of fetal/neonatal morbidity and mortality and is associated with increased offspring predisposition for cardiovascular disease (CVD) development. Mitochondria are essential organelles in maintaining cardiac function, and thus, fetal cardiac mitochondria could be responsive to the IUGR environment. In this study, we investigated whether in utero fetal cardiac mitochondrial programming can be detectable in an early stage of IUGR pregnancy. Using a well-established nonhuman IUGR primate model, we induced IUGR by reducing by 30% the maternal diet (MNR), both in males (MNR-M) and in female (MNR-F) fetuses. Fetal cardiac left ventricle (LV) tissue and blood were collected at 90 days of gestation (0.5 gestation, 0.5 G). Blood biochemical parameters were determined and heart LV mitochondrial biology assessed. MNR fetus biochemical blood parameters confirm an early fetal response to MNR. In addition, we show that in utero cardiac mitochondrial MNR adaptations are already detectable at this early stage, in a sex-divergent way. MNR induced alterations in the cardiac gene expression of oxidative phosphorylation (OXPHOS) subunits (mostly for complex-I, III, and ATP synthase), along with increased protein content for complex-I, -III, and -IV subunits only for MNR-M in comparison with male controls, highlight the fetal cardiac sex-divergent response to MNR. At this fetal stage, no major alterations were detected in mitochondrial DNA copy number nor markers for oxidative stress. This study shows that in 90-day nonhuman primate fetuses, a 30% decrease in maternal nutrition generated early in utero adaptations in fetal blood biochemical parameters and sex-specific alterations in cardiac left ventricle gene and protein expression profiles, affecting predominantly OXPHOS subunits. Since the OXPHOS system is determinant for energy production in mitochondria, our findings suggest that these early IUGR-induced mitochondrial adaptations play a role in offspring's mitochondrial dysfunction and can increase predisposition to CVD in a sex-specific way.
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- 2023
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33. Maternal hepatic adaptations during obese pregnancy encompass lobe-specific mitochondrial alterations and oxidative stress.
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Grilo LF, Martins JD, Diniz MS, Tocantins C, Cavallaro CH, Baldeiras I, Cunha-Oliveira T, Ford S, Nathanielsz PW, Oliveira PJ, and Pereira SP
- Subjects
- Humans, Female, Pregnancy, Animals, Sheep, Catalase metabolism, Liver metabolism, Oxidative Stress, Obesity metabolism, Antioxidants metabolism, Superoxide Dismutase metabolism, Glutathione metabolism, NAD metabolism, Liver Diseases metabolism
- Abstract
Maternal obesity (MO) is rising worldwide, affecting half of all gestations, constituting a possible risk-factor for some pregnancy-associated liver diseases (PALD) and hepatic diseases. PALD occur in approximately 3% of pregnancies and are characterized by maternal hepatic oxidative stress (OS) and mitochondrial dysfunction. Maternal hepatic disease increases maternal and fetal morbidity and mortality. Understanding the role of MO on liver function and pathophysiology could be crucial for better understanding the altered pathways leading to PALD and liver disease, possibly paving the way to prevention and adequate management of disease. We investigated specific hepatic metabolic alterations in mitochondria and oxidative stress during MO at late-gestation. Maternal hepatic tissue was collected at 90% gestation in Control and MO ewes (fed 150% of recommended nutrition starting 60 days before conception). Maternal hepatic redox state, mitochondrial respiratory chain (MRC), and OS markers were investigated. MO decreased MRC complex-II activity and its subunits SDHA and SDHB protein expression, increased complex-I and complex-IV activities despite reduced complex-IV subunit mtCO1 protein expression, and increased ATP synthase ATP5A subunit. Hepatic MO-metabolic remodeling was characterized by decreased adenine nucleotide translocator 1 and 2 (ANT-1/2) and voltage-dependent anion channel (VDAC) protein expression and protein kinase A (PKA) activity (P<0.01), and augmented NAD+/NADH ratio due to reduced NADH levels (P<0.01). MO showed an altered redox state with increased OS, increased lipid peroxidation (P<0.01), decreased GSH/GSSG ratio (P=0.005), increased superoxide dismutase (P=0.03) and decreased catalase (P=0.03) antioxidant enzymatic activities, lower catalase, glutathione peroxidase (GPX)-4 and glutathione reductase protein expression (P<0.05), and increased GPX-1 abundance (P=0.03). MO-related hepatic changes were more evident in the right lobe, corroborated by the integrative data analysis. Hepatic tissue from obese pregnant ewes showed alterations in the redox state, consistent with OS and MRC and metabolism remodeling. These are hallmarks of PALD and hepatic disease, supporting MO as a risk-factor and highlighting OS and mitochondrial dysfunction as mechanisms responsible for liver disease predisposition., (© 2023 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)
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- 2023
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34. Metabolic Priming as a Tool in Redox and Mitochondrial Theragnostics.
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Pinho SA, Anjo SI, and Cunha-Oliveira T
- Abstract
Theragnostics is a promising approach that integrates diagnostics and therapeutics into a single personalized strategy. To conduct effective theragnostic studies, it is essential to create an in vitro environment that accurately reflects the in vivo conditions. In this review, we discuss the importance of redox homeostasis and mitochondrial function in the context of personalized theragnostic approaches. Cells have several ways to respond to metabolic stress, including changes in protein localization, density, and degradation, which can promote cell survival. However, disruption of redox homeostasis can lead to oxidative stress and cellular damage, which are implicated in various diseases. Models of oxidative stress and mitochondrial dysfunction should be developed in metabolically conditioned cells to explore the underlying mechanisms of diseases and develop new therapies. By choosing an appropriate cellular model, adjusting cell culture conditions and validating the cellular model, it is possible to identify the most promising therapeutic options and tailor treatments to individual patients. Overall, we highlight the importance of precise and individualized approaches in theragnostics and the need to develop accurate in vitro models that reflect the in vivo conditions.
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- 2023
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35. Integrative Profiling of Amyotrophic Lateral Sclerosis Lymphoblasts Identifies Unique Metabolic and Mitochondrial Disease Fingerprints.
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Cunha-Oliveira T, Carvalho M, Sardão V, Ferreiro E, Mena D, Pereira FB, Borges F, Oliveira PJ, and Silva FSG
- Subjects
- Adenosine Triphosphate, Bayes Theorem, Humans, Mutation genetics, Superoxide Dismutase metabolism, Superoxide Dismutase-1 genetics, Amyotrophic Lateral Sclerosis genetics, Amyotrophic Lateral Sclerosis metabolism, Mitochondrial Diseases, Neurodegenerative Diseases
- Abstract
Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease with a rapid progression and no effective treatment. Metabolic and mitochondrial alterations in peripheral tissues of ALS patients may present diagnostic and therapeutic interest. We aimed to identify mitochondrial fingerprints in lymphoblast from ALS patients harboring SOD1 mutations (mutSOD1) or with unidentified mutations (undSOD1), compared with age-/sex-matched controls. Three groups of lymphoblasts, from mutSOD1 or undSOD1 ALS patients and age-/sex-matched controls, were obtained from Coriell Biobank and divided into 3 age-/sex-matched cohorts. Mitochondria-associated metabolic pathways were analyzed using Seahorse MitoStress and ATP Rate assays, complemented with metabolic phenotype microarrays, metabolite levels, gene expression, and protein expression and activity. Pooled (all cohorts) and paired (intra-cohort) analyses were performed by using bioinformatic tools, and the features with higher information gain values were selected and used for principal component analysis and Naïve Bayes classification. Considering the group as a target, the features that contributed to better segregation of control, undSOD1, and mutSOD1 were found to be the protein levels of Tfam and glycolytic ATP production rate. Metabolic phenotypic profiles in lymphoblasts from ALS patients with mutSOD1 and undSOD1 revealed unique age-dependent different substrate oxidation profiles. For most parameters, different patterns of variation in experimental endpoints in lymphoblasts were found between cohorts, which may be due to the age or sex of the donor. In the present work, we investigated several metabolic and mitochondrial hallmarks in lymphoblasts from each donor, and although a high heterogeneity of results was found, we identified specific metabolic and mitochondrial fingerprints, especially protein levels of Tfam and glycolytic ATP production rate, that may have a diagnostic and therapeutic interest., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
- Published
- 2022
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36. Mitochondrial and metabolic remodelling in human skin fibroblasts in response to glucose availability.
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Pinho SA, Costa CF, Deus CM, Pinho SLC, Miranda-Santos I, Afonso G, Bagshaw O, Stuart JA, Oliveira PJ, and Cunha-Oliveira T
- Subjects
- Adenosine Triphosphate metabolism, Energy Metabolism physiology, Fibroblasts metabolism, Humans, Oxidative Phosphorylation, Glucose metabolism, Glucose pharmacology, Mitochondria metabolism
- Abstract
Cell culture conditions highly influence cell metabolism in vitro. This is relevant for preclinical assays, for which fibroblasts are an interesting cell model, with applications in regenerative medicine, diagnostics and therapeutic development for personalized medicine, and the validation of ingredients for cosmetics. Given these cells' short lifespan in culture, we aimed to identify the best cell culture conditions and promising markers to study mitochondrial health and stress in normal human dermal fibroblasts (NHDF). We tested the effect of reducing glucose concentration in the cell medium from high glucose (HGm) to a more physiological level [low glucose medium (LGm)], or its complete removal and replacement by galactose [medium that forces oxidative phosphorylation (OXPHOSm)], always in the presence of glutamine and pyruvate. We have demonstrated that only with OXPHOSm was it possible to observe the selective inhibition of mitochondrial adenosine triphosphate (ATP) production. This reliance on mitochondrial ATP was accompanied by changes in oxygen consumption rate and extracellular acidification rate, oxidation of citric acid cycle substrates, fatty acids, lactate, and other substrates, increased mitochondrial network extension and polarization, the increased protein content of voltage-dependent anion channel (VDAC) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha and changes in several key transcripts related to energy metabolism. LGm did not promote significant metabolic changes in NHDF, although mitochondrial network extension and VDAC protein content were increased compared to HGm-cultured cells. Our results indicate that short-term adaptation to OXPHOSm is ideal for studying mitochondrial health and stress in NHDF., (© 2022 Federation of European Biochemical Societies.)
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- 2022
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37. Redox profiles of amyotrophic lateral sclerosis lymphoblasts with or without known SOD1 mutations.
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Cunha-Oliveira T, Silva DF, Segura L, Baldeiras I, Marques R, Rosenstock T, Oliveira PJ, and Silva FSG
- Subjects
- Adult, Catalase metabolism, Female, Humans, Male, Middle Aged, Mutation, NF-E2-Related Factor 2 genetics, Oxidation-Reduction, Amyotrophic Lateral Sclerosis genetics, Superoxide Dismutase-1 genetics
- Abstract
Background: Amyotrophic lateral sclerosis (ALS) is a fatal and rapidly progressing neurodegenerative disease that affects motor neurons. This disease is associated with oxidative stress especially in mutant superoxide dismutase 1 (mutSOD1) patients. However, less is known for the most prevalent sporadic ALS form, due to a lack of disease models. Here, we studied oxidative stress profiles in lymphoblasts from ALS patients with mutSOD1 or unknown (undSOD1) mutations., Methods: mutSOD1 and undSOD1 lymphoblasts, as well as sex/age-matched controls (3/group) were obtained from Coriell and divided into 46 years-old-men (C1), 46 years-old-women (C2) or 26/27 years-old-men (C3) cohorts. Growth curves were performed, and several parameters associated with redox homeostasis were evaluated, including SOD activity and expression, general oxidative stress levels, lipid peroxidation, response to oxidative stimulus, glutathione redox cycle, catalase expression, and activity, and Nrf2 transcripts. Pooled (all cohorts) and paired (intra-cohort) statistical analyses were performed, followed by clustering and principal component analyses (PCA)., Results: Although a high heterogeneity among lymphoblast redox profiles was found between cohorts, clustering analysis based on 7 parameters with high chi-square ranking (total SOD activity, oxidative stress levels, catalase transcripts, SOD1 protein levels, metabolic response to mM concentrations of tert-butyl hydroperoxide, glutathione reductase activity, and Nrf2 transcript levels) provided a perfect cluster segregation between samples from healthy controls and ALS (undSOD1 and mutSOD1), also visualized in the PCA., Conclusions: Our results show distinct redox signatures in lymphoblasts from mutSOD1, undSOD1 and healthy controls that can be used as therapeutic targets for ALS drug development., (© 2022 Stichting European Society for Clinical Investigation Journal Foundation. Published by John Wiley & Sons Ltd.)
- Published
- 2022
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38. Mitochondria-targeted anti-oxidant AntiOxCIN 4 improved liver steatosis in Western diet-fed mice by preventing lipid accumulation due to upregulation of fatty acid oxidation, quality control mechanism and antioxidant defense systems.
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Amorim R, Simões ICM, Teixeira J, Cagide F, Potes Y, Soares P, Carvalho A, Tavares LC, Benfeito S, Pereira SP, Simões RF, Karkucinska-Wieckowska A, Viegas I, Szymanska S, Dąbrowski M, Janikiewicz J, Cunha-Oliveira T, Dobrzyń A, Jones JG, Borges F, Wieckowski MR, and Oliveira PJ
- Abstract
Non-alcoholic fatty liver disease (NAFLD) is a health concern affecting 24% of the population worldwide. Although the pathophysiologic mechanisms underlying disease are not fully clarified, mitochondrial dysfunction and oxidative stress are key players in disease progression. Consequently, efforts to develop more efficient pharmacologic strategies targeting mitochondria for NAFLD prevention/treatment are underway. The conjugation of caffeic acid anti-oxidant moiety with an alkyl linker and a triphenylphosphonium cation (TPP
+ ), guided by structure-activity relationships, led to the development of a mitochondria-targeted anti-oxidant (AntiOxCIN4 ) with remarkable anti-oxidant properties. Recently, we described that AntiOxCIN4 improved mitochondrial function, upregulated anti-oxidant defense systems, and cellular quality control mechanisms (mitophagy/autophagy) via activation of the Nrf2/Keap1 pathway, preventing fatty acid-induced cell damage. Despite the data obtained, AntiOxCIN4 effects on cellular and mitochondrial energy metabolism in vivo were not studied. In the present work, we proposed that AntiOxCIN4 (2.5 mg/day/animal) may prevent non-alcoholic fatty liver (NAFL) phenotype development in a C57BL/6J mice fed with 30% high-fat, 30% high-sucrose diet for 16 weeks. HepG2 cells treated with AntiOxCIN4 (100 μM, 48 h) before the exposure to supraphysiologic free fatty acids (FFAs) (250 μM, 24 h) were used for complementary studies. AntiOxCIN4 decreased body (by 43%), liver weight (by 39%), and plasma hepatocyte damage markers in WD-fed mice. Hepatic-related parameters associated with a reduction of fat liver accumulation (by 600%) and the remodeling of fatty acyl chain composition compared with the WD-fed group were improved. Data from human HepG2 cells confirmed that a reduction of lipid droplets size and number can be a result from AntiOxCIN4 -induced stimulation of fatty acid oxidation and mitochondrial OXPHOS remodeling. In WD-fed mice, AntiOxCIN4 also induced a hepatic metabolism remodeling by upregulating mitochondrial OXPHOS, anti-oxidant defense system and phospholipid membrane composition, which is mediated by the PGC-1α-SIRT3 axis. AntiOxCIN4 prevented lipid accumulation-driven autophagic flux impairment, by increasing lysosomal proteolytic capacity. AntiOxCIN4 improved NAFL phenotype of WD-fed mice, via three main mechanisms: a) increase mitochondrial function (fatty acid oxidation); b) stimulation anti-oxidant defense system (enzymatic and non-enzymatic) and; c) prevent the impairment in autophagy. Together, the findings support the potential use of AntiOxCIN4 in the prevention/treatment of NAFLD., Competing Interests: Declaration of competing interest Paulo J. Oliveira and Fernanda Borges are cofounders of the CNC-UP spin-off company MitoTAG (Coimbra, Portugal). This spin-off had no involvement in the data collection, analysis and interpretation, manuscript writing, and the decision to submit the manuscript for publication., (Copyright © 2022. Published by Elsevier B.V.)- Published
- 2022
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39. NRF2 and Mitochondrial Function in Cancer and Cancer Stem Cells.
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Panieri E, Pinho SA, Afonso GJM, Oliveira PJ, Cunha-Oliveira T, and Saso L
- Subjects
- Humans, Kelch-Like ECH-Associated Protein 1 metabolism, Neoplastic Stem Cells metabolism, Mitochondria metabolism, NF-E2-Related Factor 2 metabolism, Neoplasms metabolism
- Abstract
The NRF2-KEAP1 system is a fundamental component of the cellular response that controls a great variety of transcriptional targets that are mainly involved in the regulation of redox homeostasis and multiple cytoprotective mechanisms that confer adaptation to the stress conditions. The pleiotropic response orchestrated by NRF2 is particularly relevant in the context of oncogenic activation, wherein this transcription factor acts as a key driver of tumor progression and cancer cells' resistance to treatment. For this reason, NRF2 has emerged as a promising therapeutic target in cancer cells, stimulating extensive research aimed at the identification of natural, as well as chemical, NRF2 inhibitors. Excitingly, the influence of NRF2 on cancer cells' biology extends far beyond its mere antioxidant function and rather encompasses a functional crosstalk with the mitochondrial network that can influence crucial aspects of mitochondrial homeostasis, including biogenesis, oxidative phosphorylation, metabolic reprogramming, and mitophagy. In the present review, we summarize the current knowledge of the reciprocal interrelation between NRF2 and mitochondria, with a focus on malignant tumors and cancer stem cells.
- Published
- 2022
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40. Evaluation of 6-Hydroxydopamine and Rotenone In Vitro Neurotoxicity on Differentiated SH-SY5Y Cells Using Applied Computational Statistics.
- Author
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Simões RF, Oliveira PJ, Cunha-Oliveira T, and Pereira FB
- Subjects
- Apoptosis, Cell Death, Cell Line, Tumor, Cell Survival, Humans, Oxidopamine toxicity, Rotenone toxicity, Neuroprotective Agents pharmacology, Neurotoxicity Syndromes etiology
- Abstract
With the increase in life expectancy and consequent aging of the world's population, the prevalence of many neurodegenerative diseases is increasing, without concomitant improvement in diagnostics and therapeutics. These diseases share neuropathological hallmarks, including mitochondrial dysfunction. In fact, as mitochondrial alterations appear prior to neuronal cell death at an early phase of a disease's onset, the study and modulation of mitochondrial alterations have emerged as promising strategies to predict and prevent neurotoxicity and neuronal cell death before the onset of cell viability alterations. In this work, differentiated SH-SY5Y cells were treated with the mitochondrial-targeted neurotoxicants 6-hydroxydopamine and rotenone. These compounds were used at different concentrations and for different time points to understand the similarities and differences in their mechanisms of action. To accomplish this, data on mitochondrial parameters were acquired and analyzed using unsupervised (hierarchical clustering) and supervised (decision tree) machine learning methods. Both biochemical and computational analyses resulted in an evident distinction between the neurotoxic effects of 6-hydroxydopamine and rotenone, specifically for the highest concentrations of both compounds.
- Published
- 2022
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41. Quantitative analysis of neuronal mitochondrial movement reveals patterns resulting from neurotoxicity of rotenone and 6-hydroxydopamine.
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Simões RF, Pino R, Moreira-Soares M, Kovarova J, Neuzil J, Travasso R, Oliveira PJ, Cunha-Oliveira T, and Pereira FB
- Subjects
- Adrenergic Agents adverse effects, Cell Differentiation, Humans, Mitochondria drug effects, Neuroblastoma chemically induced, Neurons drug effects, Uncoupling Agents adverse effects, Mitochondria pathology, Mitochondrial Dynamics, Neuroblastoma pathology, Neurons pathology, Oxidopamine adverse effects, Rotenone adverse effects
- Abstract
Alterations in mitochondrial dynamics, including their intracellular trafficking, are common early manifestations of neuronal degeneration. However, current methodologies used to study mitochondrial trafficking events rely on parameters that are primarily altered in later stages of neurodegeneration. Our objective was to establish a reliable applied statistical analysis to detect early alterations in neuronal mitochondrial trafficking. We propose a novel quantitative analysis of mitochondria trajectories based on innovative movement descriptors, including straightness, efficiency, anisotropy, and kurtosis. We evaluated time- and dose-dependent alterations in trajectory descriptors using biological data from differentiated SH-SY5Y cells treated with the mitochondrial toxicants 6-hydroxydopamine and rotenone. MitoTracker Red CMXRos-labelled mitochondria movement was analyzed by total internal reflection fluorescence microscopy followed by computational modelling to describe the process. Based on the aforementioned trajectory descriptors, this innovative analysis of mitochondria trajectories provides insights into mitochondrial movement characteristics and can be a consistent and sensitive method to detect alterations in mitochondrial trafficking occurring in the earliest time points of neurodegeneration., (© 2021 Federation of American Societies for Experimental Biology.)
- Published
- 2021
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42. Corrigendum to 'A mitochondria-targeted caffeic acid derivative reverts cellular and mitochondrial defects in human skin fibroblasts from male sporadic Parkinson's disease patients'. [Redox Biology 45 (2021) 102037].
- Author
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Deus CM, Pereira SP, Cunha-Oliveira T, Teixeira J, Simões RF, Cagide F, Benfeito S, Borges F, Raimundo N, and Oliveira PJ
- Published
- 2021
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43. A mitochondria-targeted caffeic acid derivative reverts cellular and mitochondrial defects in human skin fibroblasts from male sporadic Parkinson's disease patients.
- Author
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Deus CM, Pereira SP, Cunha-Oliveira T, Teixeira J, Simões RF, Cagide F, Benfeito S, Borges F, Raimundo N, and Oliveira PJ
- Subjects
- Caffeic Acids metabolism, Fibroblasts metabolism, Humans, Male, Mitochondria metabolism, Oxidative Stress, Parkinson Disease drug therapy, Parkinson Disease metabolism
- Abstract
Parkinson's Disease (PD) is a neurodegenerative disorder affecting more than 10 million people worldwide. Currently, PD has no cure and no early diagnostics methods exist. Mitochondrial dysfunction is presented in the early stages of PD, and it is considered an important pathophysiology component. We have previously developed mitochondria-targeted hydroxycinnamic acid derivatives, presenting antioxidant and iron-chelating properties, and preventing oxidative stress in several biological models of disease. We have also demonstrated that skin fibroblasts from male sporadic PD patients (sPD) presented cellular and mitochondrial alterations, including increased oxidative stress, hyperpolarized and elongated mitochondria and decreased respiration and ATP levels. We also showed that forcing mitochondrial oxidative phosphorylation (OXPHOS) in sPD fibroblasts uncovers metabolic defects that were otherwise hidden. In this work, we tested the hypothesis that a lead mitochondria-targeted hydroxycinnamic acid derivative would revert the phenotype found in skin fibroblasts from sPD patients. Our results demonstrated that treating human skin fibroblasts from sPD patients with non-toxic concentrations of AntiOxCIN
4 restored mitochondrial membrane potential and mitochondrial fission, decreased autophagic flux, and enhanced cellular responses to stress by improving the cellular redox state and decreasing reactive oxygen species (ROS) levels. Besides, fibroblasts from sPD patients treated with AntiOxCIN4 showed increased maximal respiration and metabolic activity, converting sPD fibroblasts physiologically more similar to their sex- and age-matched healthy controls. The positive compound effect was reinforced using a supervised machine learning model, confirming that AntiOxCIN4 treatment converted treated fibroblasts from sPD patients closer to the phenotype of control fibroblasts. Our data points out a possible mechanism of AntiOxCIN4 action contributing to a deeper understanding of how the use of mitochondria-targeted antioxidants based on a polyphenol scaffold can be used as potential drug candidates for delaying PD progression, validating the use of fibroblasts from sPD patients with more active OXPHOS as platforms for mitochondria-based drug development., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2021
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44. Sex-dependent vulnerability of fetal nonhuman primate cardiac mitochondria to moderate maternal nutrient reduction.
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Pereira SP, Tavares LC, Duarte AI, Baldeiras I, Cunha-Oliveira T, Martins JD, Santos MS, Maloyan A, Moreno AJ, Cox LA, Li C, Nathanielsz PW, Nijland MJ, and Oliveira PJ
- Subjects
- Adenine Nucleotides metabolism, Animals, Female, Fetal Nutrition Disorders pathology, Mitochondria, Heart ultrastructure, Oxidative Stress, Papio, Pregnancy, Fetal Nutrition Disorders metabolism, Maternal Nutritional Physiological Phenomena, Mitochondria, Heart metabolism
- Abstract
Poor maternal nutrition in pregnancy affects fetal development, predisposing offspring to cardiometabolic diseases. The role of mitochondria during fetal development on later-life cardiac dysfunction caused by maternal nutrient reduction (MNR) remains unexplored. We hypothesized that MNR during gestation causes fetal cardiac bioenergetic deficits, compromising cardiac mitochondrial metabolism and reserve capacity. To enable human translation, we developed a primate baboon model (Papio spp.) of moderate MNR in which mothers receive 70% of control nutrition during pregnancy, resulting in intrauterine growth restriction (IUGR) offspring and later exhibiting myocardial remodeling and heart failure at human equivalent ∼25 years. Term control and MNR baboon offspring were necropsied following cesarean-section, and left ventricle (LV) samples were collected. MNR adversely impacted fetal cardiac LV mitochondria in a sex-dependent fashion. Increased maternal plasma aspartate aminotransferase, creatine phosphokinase (CPK), and elevated cortisol levels in MNR concomitant with decreased blood insulin in male fetal MNR were measured. MNR resulted in a two-fold increase in fetal LV mitochondrial DNA (mtDNA). MNR resulted in increased transcripts for several respiratory chain (NDUFB8, UQCRC1, and cytochrome c) and adenosine triphosphate (ATP) synthase proteins. However, MNR fetal LV mitochondrial complex I and complex II/III activities were significantly decreased, possibly contributing to the 73% decreased ATP content and increased lipid peroxidation. MNR fetal LV showed mitochondria with sparse and disarranged cristae dysmorphology. Conclusion: MNR disruption of fetal cardiac mitochondrial fitness likely contributes to the documented developmental programming of adult cardiac dysfunction, indicating a programmed mitochondrial inability to deliver sufficient energy to cardiac tissues as a chronic mechanism for later-life heart failure., (© 2021 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)
- Published
- 2021
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45. MPV17 Mutations Are Associated With a Quiescent Energetic Metabolic Profile.
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Jacinto S, Guerreiro P, de Oliveira RM, Cunha-Oliveira T, Santos MJ, Grazina M, Rego AC, and Outeiro TF
- Abstract
Mutations in the MPV17 gene are associated with hepatocerebral form of mitochondrial depletion syndrome. The mechanisms through which MPV17 mutations cause respiratory chain dysfunction and mtDNA depletion is still unclear. The MPV17 gene encodes an inner membrane mitochondrial protein that was recently described to function as a non-selective channel. Although its exact function is unknown, it is thought to be important in the maintenance of mitochondrial membrane potential (ΔΨm). To obtain more information about the role of MPV17 in human disease, we investigated the effect of MPV17 knockdown and of selected known MPV17 mutations associated with MPV17 disease in vitro . We used different approaches in order to evaluate the cellular consequences of MPV17 deficiency. We found that lower levels of MPV17 were associated with impaired mitochondrial respiration and with a quiescent energetic metabolic profile. All the mutations studied destabilized the protein, resulting in reduced protein levels. We also demonstrated that different mutations caused different cellular abnormalities, including increased ROS production, decreased oxygen consumption, loss of ΔΨm, and mislocalization of MPV17 protein. Our study provides novel insight into the molecular effects of MPV17 mutations and opens novel possibilities for testing therapeutic strategies for a devastating group of disorders., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Jacinto, Guerreiro, de Oliveira, Cunha-Oliveira, Santos, Grazina, Rego and Outeiro.)
- Published
- 2021
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46. Refinement of a differentiation protocol using neuroblastoma SH-SY5Y cells for use in neurotoxicology research.
- Author
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Simões RF, Ferrão R, Silva MR, Pinho SLC, Ferreira L, Oliveira PJ, and Cunha-Oliveira T
- Subjects
- Cell Line, Tumor, Colorimetry, Humans, Microscopy methods, Mitochondria drug effects, Mitochondria metabolism, Neurons drug effects, Neurons physiology, Rhodamines, Cell Differentiation, Neuroblastoma, Neurotoxicity Syndromes pathology, Tretinoin toxicity
- Abstract
Since most models used to study neuronal dysfunction display disadvantages and ethical concerns, a fast and reproducible in vitro model to study mitochondria-related neurodegeneration is required. Here, we optimized and characterized a 3-day retinoic acid-based protocol to differentiate the SH-SY5Y cell line into a neuronal-like phenotype and investigated alterations in mitochondrial physiology and distribution. Differentiation was associated with p21-linked cell cycle arrest and an increase in cell mass and area, possibly associated with the development of neurite-like extensions. Notably, increased expression of mature neuronal markers (neuronal-specific nuclear protein, microtubule-associated protein 2, βIII tubulin and enolase 2) was observed in differentiated cells. Moreover, increased mitochondrial content and maximal area per cell suggests mitochondrial remodeling. To demonstrate that this model is appropriate to study mitochondrial dysfunction, cells were treated for 6 h with mitochondrial toxicants (rotenone, antimycin A, carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) and 6-hydroxydopamine (6-OHDA)). Differentiated cells were more susceptible to increasing concentrations of FCCP, antimycin A, and rotenone, while 6-OHDA showed a distinct dose-dependent neurotoxicity pattern. Even though differentiated cells did not exhibit a fully mature/differentiated neuronal phenotype, the protocol developed can be used to study neurotoxicity processes, mitochondrial dynamics, and bioenergetic impairment, representing an alternative to study mitochondrial impairment-related pathologies in vitro., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
- Published
- 2021
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47. Maternal obesity in sheep impairs foetal hepatic mitochondrial respiratory chain capacity.
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Serafim TL, Cunha-Oliveira T, Deus CM, Sardão VA, Cardoso IM, Yang S, Odhiambo JF, Ghnenis AB, Smith AM, Li J, Nathanielsz PW, Ford SP, and Oliveira PJ
- Subjects
- Animals, Beclin-1 metabolism, Cardiolipins metabolism, Cathepsin B metabolism, Cathepsin D metabolism, Female, Microtubule-Associated Proteins metabolism, Phospholipids metabolism, Pregnancy, Sheep, Superoxide Dismutase metabolism, Autophagy physiology, Electron Transport physiology, Fetus metabolism, Liver metabolism, Mitochondria, Liver metabolism, Obesity, Maternal metabolism
- Abstract
Background: Changes in the nutritional environment in utero induced by maternal obesity (MO) lead to foetal metabolic dysfunction predisposing offspring to later-life metabolic diseases. Since mitochondria play a crucial role in hepatic metabolism and function, we hypothesized that MO prior to conception and throughout pregnancy programmes foetal sheep liver mitochondrial phenotype., Material and Methods: Ewes ate an obesogenic diet (150% requirements; MO), or 100% requirements (CTR), from 60 days prior to conception. Foetal livers were removed at 0.9 gestation. We measured foetal liver mitochondrial DNA copy number, activity of superoxide dismutase, cathepsins B and D and selected protein content, total phospholipids and cardiolipin and activity of mitochondrial respiratory chain complexes., Results: A significant decrease in activities of mitochondrial complexes I, II-III and IV, but not aconitase, was observed in MO. In the antioxidant machinery, there was a significant increase in activity of total superoxide dismutase (SOD) and SOD2 in MO. However, no differences were found regarding autophagy-related protein content (p62, beclin-I, LC3-I, LC3-II and Lamp2A) and cathepsin B and D activities. A 21.5% decrease in total mitochondrial phospholipid was observed in MO., Conclusions: The data indicate that MO impairs foetal hepatic mitochondrial oxidative capacity and affects total mitochondrial phospholipid content. In addition, MO affects the regulation of foetal liver redox pathways, indicating metabolic adaptations to the higher foetal lipid environment. Consequences of in utero programming of foetal hepatic metabolism may persist and compromise mitochondrial bioenergetics in later life, and increase susceptibility to metabolic diseases., (© 2020 Stichting European Society for Clinical Investigation Journal Foundation. Published by John Wiley & Sons Ltd.)
- Published
- 2021
- Full Text
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48. Analysis of Proapoptotic Protein Trafficking to and from Mitochondria.
- Author
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Vega-Naredo I, Oliveira G, Cunha-Oliveira T, Serafim T, Sardão VA, and Oliveira PJ
- Subjects
- Animals, Cell Fractionation, Cell Line, Centrifugation, Humans, Microscopy, Fluorescence, Mitochondria pathology, Protein Transport, Apoptosis, Apoptosis Regulatory Proteins metabolism, Blotting, Western, Fluorescent Antibody Technique, Immunoprecipitation, Mitochondria metabolism
- Abstract
Mitochondria play a key role in cell death and its regulation. The permeabilization of the outer mitochondrial membrane, which is mainly controlled by proteins of the BCL-2 family, is a key event that can be directly induced by different signaling pathways, including p53-mediated, and results in the release of proapoptotic factors to the cytosol, such as cytochrome c, second mitochondria-derived activator of caspases/direct inhibitor-of-apoptosis (IAP) binding protein with low pI (SMAC/Diablo), Omi serine protease (Omi/HtrA2), apoptosis-inducing factor (AIF), or endonuclease G (Endo-G). Hence, the determination of subcellular localization of these proteins is extremely important to predict cell fate and elucidate the specific mechanism of apoptosis. Here we describe experimental protocols that can be used to study the subcellular location of different proapoptotic proteins to be used in basic cell biology and toxicology studies.
- Published
- 2021
- Full Text
- View/download PDF
49. Oxidative Stress in Amyotrophic Lateral Sclerosis: Pathophysiology and Opportunities for Pharmacological Intervention.
- Author
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Cunha-Oliveira T, Montezinho L, Mendes C, Firuzi O, Saso L, Oliveira PJ, and Silva FSG
- Subjects
- Animals, Disease Models, Animal, Humans, Amyotrophic Lateral Sclerosis drug therapy, Amyotrophic Lateral Sclerosis metabolism, Amyotrophic Lateral Sclerosis pathology, Antioxidants therapeutic use, Motor Neurons metabolism, Motor Neurons pathology, Neuroprotective Agents therapeutic use, Oxidative Stress drug effects
- Abstract
Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig's disease or Charcot disease, is a fatal neurodegenerative disease that affects motor neurons (MNs) and leads to death within 2-5 years of diagnosis, without any effective therapy available. Although the pathological mechanisms leading to ALS are still unknown, a wealth of evidence indicates that an excessive reactive oxygen species (ROS) production associated with an inefficient antioxidant defense represents an important pathological feature in ALS. Substantial evidence indicates that oxidative stress (OS) is implicated in the loss of MNs and in mitochondrial dysfunction, contributing decisively to neurodegeneration in ALS. Although the modulation of OS represents a promising approach to protect MNs from degeneration, the fact that several antioxidants with beneficial effects in animal models failed to show any therapeutic benefit in patients raises several questions that should be analyzed. Using specific queries for literature search on PubMed, we review here the role of OS-related mechanisms in ALS, including the involvement of altered mitochondrial function with repercussions in neurodegeneration. We also describe antioxidant compounds that have been mostly tested in preclinical and clinical trials of ALS, also describing their respective mechanisms of action. While the description of OS mechanism in the different mutations identified in ALS has as principal objective to clarify the contribution of OS in ALS, the description of positive and negative outcomes for each antioxidant is aimed at paving the way for novel opportunities for intervention. In conclusion, although antioxidant strategies represent a very promising approach to slow the progression of the disease, it is of utmost need to invest on the characterization of OS profiles representative of each subtype of patient, in order to develop personalized therapies, allowing to understand the characteristics of antioxidants that have beneficial effects on different subtypes of patients., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 Teresa Cunha-Oliveira et al.)
- Published
- 2020
- Full Text
- View/download PDF
50. Mitochondrial remodeling in human skin fibroblasts from sporadic male Parkinson's disease patients uncovers metabolic and mitochondrial bioenergetic defects.
- Author
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Deus CM, Pereira SP, Cunha-Oliveira T, Pereira FB, Raimundo N, and Oliveira PJ
- Subjects
- Aged, Galactose metabolism, Glucose metabolism, Humans, Male, Middle Aged, Oxidative Phosphorylation, Oxygen Consumption physiology, Pyruvic Acid metabolism, Substantia Nigra metabolism, Energy Metabolism physiology, Fibroblasts metabolism, Metabolic Diseases metabolism, Mitochondria metabolism, Mitochondrial Diseases metabolism, Parkinson Disease metabolism, Skin metabolism
- Abstract
Parkinson's Disease (PD) is characterized by dopaminergic neurodegeneration in the substantia nigra. The exact mechanism by which dopaminergic neurodegeneration occurs is still unknown; however, mitochondrial dysfunction has long been implicated in PD pathogenesis. To investigate the sub-cellular events that lead to disease progression and to develop personalized interventions, non-neuronal cells which are collected in a minimally invasive manner can be key to test interventions aimed at improving mitochondrial function. We used human skin fibroblasts from sporadic PD (sPD) patients as a cell proxy to detect metabolic and mitochondrial alterations which would also exist in a non-neuronal cell type. In this model, we used a glucose-free/galactose- glutamine- and pyruvate-containing cell culture medium, which forces cells to be more dependent on oxidative phosphorylation (OXPHOS) for energy production, in order to reveal hidden metabolic and mitochondrial alterations present in fibroblasts from sPD patients. We demonstrated that fibroblasts from sPD patients show hyperpolarized and elongated mitochondrial networks and higher mitochondrial ROS concentration, as well as decreased ATP levels and glycolysis-related ECAR. Our results also showed that abnormalities of fibroblasts from sPD patients became more evident when stimulating OXPHOS. Under these culture conditions, fibroblasts from sPD cells presented decreased basal respiration, ATP-linked OCR and maximal respiration, and increased mitochondria-targeting phosphorylation of DRP1 when compared to control cells. Our work validates the relevance of using fibroblasts from sPD patients to study cellular and molecular changes that are characteristic of dopaminergic neurodegeneration of PD, and shows that forcing mitochondrial OXPHOS uncovers metabolic defects that were otherwise hidden., (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Published
- 2020
- Full Text
- View/download PDF
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