Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells

Silvia Magalhães-Novais; Juan C. Bermejo-Millo; Rute Loureiro; Katia A. Mesquita; M. Rosário Domingues; Elisabete Maciel; Tânia Melo; Inês Baldeiras; Jenna R. Erickson; Jon M Holy; Yaiza Potes; Ana Coto-Montes; Paulo J. Oliveira; Ignacio Vega-Naredo

doi:10.1080/15548627.2019.1607694

Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells

Silvia Magalhães-Novais, Juan C. Bermejo-Millo, Rute Loureiro, Katia A. Mesquita, M. Rosário Domingues, Elisabete Maciel, Tânia Melo, Inês Baldeiras, Jenna R. Erickson, Jon M Holy, Yaiza Potes, Ana Coto-Montes, Paulo J. Oliveira, Ignacio Vega-Naredo

Biomedical Sciences

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14 Scopus citations

Abstract

Given the relatively long life of stem cells (SCs), efficient mechanisms of quality control to balance cell survival and resistance to external and internal stress are required. Our objective was to test the relevance of cell quality control mechanisms for SCs maintenance, differentiation and resistance to cell death. We compared cell quality control in P19 stem cells (P19SCs) before and after differentiation (P19dCs). Differentiation of P19SCs resulted in alterations in parameters involved in cell survival and protein homeostasis, including the redox system, cardiolipin and lipid profiles, unfolded protein response, ubiquitin-proteasome and lysosomal systems, and signaling pathways controlling cell growth. In addition, P19SCs pluripotency was correlated with stronger antioxidant protection, modulation of apoptosis, and activation of macroautophagy, which all contributed to preserve SCs quality by increasing the threshold for cell death activation. Furthermore, our findings identify critical roles for the PI3K-AKT-MTOR pathway, as well as autophagic flux and apoptosis regulation in the maintenance of P19SCs pluripotency and differentiation potential. Abbreviations: 3-MA: 3-methyladenine; AKT/protein kinase B: thymoma viral proto-oncogene; AKT1: thymoma viral proto-oncogene 1; ATG: AuTophaGy-related; ATF6: activating transcription factor 6; BAX: BCL2-associated X protein; BBC3/PUMA: BCL2 binding component 3; BCL2: B cell leukemia/lymphoma 2; BNIP3L: BCL2/adenovirus E1B interacting protein 3-like; CASP3: caspase 3; CASP8: caspase 8; CASP9: caspase 9; CL: cardiolipin; CTSB: cathepsin B; CTSD: cathepsin D; DDIT3/CHOP: DNA-damage inducible transcript 3; DNM1L/DRP1: dynamin 1-like; DRAM1: DNA-damage regulated autophagy modulator 1; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; EIF2S1/eIF2α: eukaryotic translation initiation factor 2, subunit alpha; ERN1/IRE1α: endoplasmic reticulum to nucleus signaling 1; ESCs: embryonic stem cells; KRT8/TROMA-1: cytokeratin 8; LAMP2A: lysosomal-associated membrane protein 2A; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; NANOG: Nanog homeobox; NAO: 10-N-nonyl acridine orange; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; OPA1: OPA1, mitochondrial dynamin like GTPase; P19dCs: P19 differentiated cells; P19SCs: P19 stem cells; POU5F1/OCT4: POU domain, class 5, transcription factor 1; PtdIns3K: phosphatidylinositol 3-kinase; RA: retinoic acid; ROS: reactive oxygen species; RPS6KB1/p70S6K: ribosomal protein S6 kinase, polypeptide 1; SCs: stem cells; SOD: superoxide dismutase; SHC1-1/p66SHC: src homology 2 domain-containing transforming protein C1, 66 kDa isoform; SOX2: SRY (sex determining region Y)-box 2; SQSTM1/p62: sequestosome 1; SPTAN1/αII-spectrin: spectrin alpha, non-erythrocytic 1; TOMM20: translocase of outer mitochondrial membrane 20; TRP53/p53: transformation related protein 53; TUBB3/betaIII-tubulin: tubulin, beta 3 class III; UPR: unfolded protein response; UPS: ubiquitin-proteasome system.

Original language	English (US)
Pages (from-to)	313-333
Number of pages	21
Journal	Autophagy
Volume	16
Issue number	2
DOIs	https://doi.org/10.1080/15548627.2019.1607694
State	Published - Feb 1 2020

Bibliographical note

Funding Information:
This work was supported by the European Regional Development Fund [POCI-01-0145-FEDER-016390:CANCEL STEM];European Regional Development Fund [POCI-01-0145-FEDER-007440];Fundação para a Ciência e a Tecnologia [IF/01316/2014];Instituto de Salud Carlos III [FI14/00405];Instituto de Salud Carlos III [PI17/02009];Instituto de Investigación Sanitaria del Principado de Asturias. This work was supported by the European Regional Development Fund (ERDF) through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT–Fundação para a Ciência e a Tecnologia, under grants POCI-01-0145-FEDER-016390:CANCEL STEM, POCI-01-0145-FEDER-007440, and IF/01316/2014; the Spanish Ministry of Science, Innovation and Universities-ISCIII under grant PI17/02009. J.C. B-M. acknowledges his predoctoral fellow from the Instituto de Investigación Sanitaria del Principado de Asturias (ISPA). Y.P. acknowledges her predoctoral fellow (FI14/00405) from the Instituto de Salud Carlos III (Spanish Ministry of Science, Innovation and Universities).

Publisher Copyright:
© 2019, © 2019 Informa UK Limited, trading as Taylor & Francis Group.

Keywords

Autophagy
cell death
metabolism
redox system
stem cells differentiation

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10.1080/15548627.2019.1607694

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Magalhães-Novais, S., Bermejo-Millo, J. C., Loureiro, R., Mesquita, K. A., Domingues, M. R., Maciel, E., Melo, T., Baldeiras, I., Erickson, J. R., Holy, J. M., Potes, Y., Coto-Montes, A., Oliveira, P. J., & Vega-Naredo, I. (2020). Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells. Autophagy, 16(2), 313-333. https://doi.org/10.1080/15548627.2019.1607694

Magalhães-Novais, S, Bermejo-Millo, JC, Loureiro, R, Mesquita, KA, Domingues, MR, Maciel, E, Melo, T, Baldeiras, I, Erickson, JR, Holy, JM, Potes, Y, Coto-Montes, A, Oliveira, PJ & Vega-Naredo, I 2020, 'Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells', Autophagy, vol. 16, no. 2, pp. 313-333. https://doi.org/10.1080/15548627.2019.1607694

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title = "Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells",

abstract = "Given the relatively long life of stem cells (SCs), efficient mechanisms of quality control to balance cell survival and resistance to external and internal stress are required. Our objective was to test the relevance of cell quality control mechanisms for SCs maintenance, differentiation and resistance to cell death. We compared cell quality control in P19 stem cells (P19SCs) before and after differentiation (P19dCs). Differentiation of P19SCs resulted in alterations in parameters involved in cell survival and protein homeostasis, including the redox system, cardiolipin and lipid profiles, unfolded protein response, ubiquitin-proteasome and lysosomal systems, and signaling pathways controlling cell growth. In addition, P19SCs pluripotency was correlated with stronger antioxidant protection, modulation of apoptosis, and activation of macroautophagy, which all contributed to preserve SCs quality by increasing the threshold for cell death activation. Furthermore, our findings identify critical roles for the PI3K-AKT-MTOR pathway, as well as autophagic flux and apoptosis regulation in the maintenance of P19SCs pluripotency and differentiation potential. Abbreviations: 3-MA: 3-methyladenine; AKT/protein kinase B: thymoma viral proto-oncogene; AKT1: thymoma viral proto-oncogene 1; ATG: AuTophaGy-related; ATF6: activating transcription factor 6; BAX: BCL2-associated X protein; BBC3/PUMA: BCL2 binding component 3; BCL2: B cell leukemia/lymphoma 2; BNIP3L: BCL2/adenovirus E1B interacting protein 3-like; CASP3: caspase 3; CASP8: caspase 8; CASP9: caspase 9; CL: cardiolipin; CTSB: cathepsin B; CTSD: cathepsin D; DDIT3/CHOP: DNA-damage inducible transcript 3; DNM1L/DRP1: dynamin 1-like; DRAM1: DNA-damage regulated autophagy modulator 1; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; EIF2S1/eIF2α: eukaryotic translation initiation factor 2, subunit alpha; ERN1/IRE1α: endoplasmic reticulum to nucleus signaling 1; ESCs: embryonic stem cells; KRT8/TROMA-1: cytokeratin 8; LAMP2A: lysosomal-associated membrane protein 2A; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; NANOG: Nanog homeobox; NAO: 10-N-nonyl acridine orange; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; OPA1: OPA1, mitochondrial dynamin like GTPase; P19dCs: P19 differentiated cells; P19SCs: P19 stem cells; POU5F1/OCT4: POU domain, class 5, transcription factor 1; PtdIns3K: phosphatidylinositol 3-kinase; RA: retinoic acid; ROS: reactive oxygen species; RPS6KB1/p70S6K: ribosomal protein S6 kinase, polypeptide 1; SCs: stem cells; SOD: superoxide dismutase; SHC1-1/p66SHC: src homology 2 domain-containing transforming protein C1, 66 kDa isoform; SOX2: SRY (sex determining region Y)-box 2; SQSTM1/p62: sequestosome 1; SPTAN1/αII-spectrin: spectrin alpha, non-erythrocytic 1; TOMM20: translocase of outer mitochondrial membrane 20; TRP53/p53: transformation related protein 53; TUBB3/betaIII-tubulin: tubulin, beta 3 class III; UPR: unfolded protein response; UPS: ubiquitin-proteasome system.",

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author = "Silvia Magalh{\~a}es-Novais and Bermejo-Millo, {Juan C.} and Rute Loureiro and Mesquita, {Katia A.} and Domingues, {M. Ros{\'a}rio} and Elisabete Maciel and T{\^a}nia Melo and In{\^e}s Baldeiras and Erickson, {Jenna R.} and Holy, {Jon M} and Yaiza Potes and Ana Coto-Montes and Oliveira, {Paulo J.} and Ignacio Vega-Naredo",

note = "Funding Information: This work was supported by the European Regional Development Fund [POCI-01-0145-FEDER-016390:CANCEL STEM];European Regional Development Fund [POCI-01-0145-FEDER-007440];Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia [IF/01316/2014];Instituto de Salud Carlos III [FI14/00405];Instituto de Salud Carlos III [PI17/02009];Instituto de Investigaci{\'o}n Sanitaria del Principado de Asturias. This work was supported by the European Regional Development Fund (ERDF) through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT–Funda{\c c}{\~a}o para a Ci{\^e}ncia e a Tecnologia, under grants POCI-01-0145-FEDER-016390:CANCEL STEM, POCI-01-0145-FEDER-007440, and IF/01316/2014; the Spanish Ministry of Science, Innovation and Universities-ISCIII under grant PI17/02009. J.C. B-M. acknowledges his predoctoral fellow from the Instituto de Investigaci{\'o}n Sanitaria del Principado de Asturias (ISPA). Y.P. acknowledges her predoctoral fellow (FI14/00405) from the Instituto de Salud Carlos III (Spanish Ministry of Science, Innovation and Universities). Publisher Copyright: {\textcopyright} 2019, {\textcopyright} 2019 Informa UK Limited, trading as Taylor & Francis Group.",

year = "2020",

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doi = "10.1080/15548627.2019.1607694",

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T1 - Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells

AU - Magalhães-Novais, Silvia

AU - Bermejo-Millo, Juan C.

AU - Loureiro, Rute

AU - Mesquita, Katia A.

AU - Domingues, M. Rosário

AU - Maciel, Elisabete

AU - Melo, Tânia

AU - Baldeiras, Inês

AU - Erickson, Jenna R.

AU - Holy, Jon M

AU - Potes, Yaiza

AU - Coto-Montes, Ana

AU - Oliveira, Paulo J.

AU - Vega-Naredo, Ignacio

N1 - Funding Information: This work was supported by the European Regional Development Fund [POCI-01-0145-FEDER-016390:CANCEL STEM];European Regional Development Fund [POCI-01-0145-FEDER-007440];Fundação para a Ciência e a Tecnologia [IF/01316/2014];Instituto de Salud Carlos III [FI14/00405];Instituto de Salud Carlos III [PI17/02009];Instituto de Investigación Sanitaria del Principado de Asturias. This work was supported by the European Regional Development Fund (ERDF) through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation and Portuguese national funds via FCT–Fundação para a Ciência e a Tecnologia, under grants POCI-01-0145-FEDER-016390:CANCEL STEM, POCI-01-0145-FEDER-007440, and IF/01316/2014; the Spanish Ministry of Science, Innovation and Universities-ISCIII under grant PI17/02009. J.C. B-M. acknowledges his predoctoral fellow from the Instituto de Investigación Sanitaria del Principado de Asturias (ISPA). Y.P. acknowledges her predoctoral fellow (FI14/00405) from the Instituto de Salud Carlos III (Spanish Ministry of Science, Innovation and Universities). Publisher Copyright: © 2019, © 2019 Informa UK Limited, trading as Taylor & Francis Group.

PY - 2020/2/1

Y1 - 2020/2/1

N2 - Given the relatively long life of stem cells (SCs), efficient mechanisms of quality control to balance cell survival and resistance to external and internal stress are required. Our objective was to test the relevance of cell quality control mechanisms for SCs maintenance, differentiation and resistance to cell death. We compared cell quality control in P19 stem cells (P19SCs) before and after differentiation (P19dCs). Differentiation of P19SCs resulted in alterations in parameters involved in cell survival and protein homeostasis, including the redox system, cardiolipin and lipid profiles, unfolded protein response, ubiquitin-proteasome and lysosomal systems, and signaling pathways controlling cell growth. In addition, P19SCs pluripotency was correlated with stronger antioxidant protection, modulation of apoptosis, and activation of macroautophagy, which all contributed to preserve SCs quality by increasing the threshold for cell death activation. Furthermore, our findings identify critical roles for the PI3K-AKT-MTOR pathway, as well as autophagic flux and apoptosis regulation in the maintenance of P19SCs pluripotency and differentiation potential. Abbreviations: 3-MA: 3-methyladenine; AKT/protein kinase B: thymoma viral proto-oncogene; AKT1: thymoma viral proto-oncogene 1; ATG: AuTophaGy-related; ATF6: activating transcription factor 6; BAX: BCL2-associated X protein; BBC3/PUMA: BCL2 binding component 3; BCL2: B cell leukemia/lymphoma 2; BNIP3L: BCL2/adenovirus E1B interacting protein 3-like; CASP3: caspase 3; CASP8: caspase 8; CASP9: caspase 9; CL: cardiolipin; CTSB: cathepsin B; CTSD: cathepsin D; DDIT3/CHOP: DNA-damage inducible transcript 3; DNM1L/DRP1: dynamin 1-like; DRAM1: DNA-damage regulated autophagy modulator 1; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; EIF2S1/eIF2α: eukaryotic translation initiation factor 2, subunit alpha; ERN1/IRE1α: endoplasmic reticulum to nucleus signaling 1; ESCs: embryonic stem cells; KRT8/TROMA-1: cytokeratin 8; LAMP2A: lysosomal-associated membrane protein 2A; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; NANOG: Nanog homeobox; NAO: 10-N-nonyl acridine orange; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; OPA1: OPA1, mitochondrial dynamin like GTPase; P19dCs: P19 differentiated cells; P19SCs: P19 stem cells; POU5F1/OCT4: POU domain, class 5, transcription factor 1; PtdIns3K: phosphatidylinositol 3-kinase; RA: retinoic acid; ROS: reactive oxygen species; RPS6KB1/p70S6K: ribosomal protein S6 kinase, polypeptide 1; SCs: stem cells; SOD: superoxide dismutase; SHC1-1/p66SHC: src homology 2 domain-containing transforming protein C1, 66 kDa isoform; SOX2: SRY (sex determining region Y)-box 2; SQSTM1/p62: sequestosome 1; SPTAN1/αII-spectrin: spectrin alpha, non-erythrocytic 1; TOMM20: translocase of outer mitochondrial membrane 20; TRP53/p53: transformation related protein 53; TUBB3/betaIII-tubulin: tubulin, beta 3 class III; UPR: unfolded protein response; UPS: ubiquitin-proteasome system.

AB - Given the relatively long life of stem cells (SCs), efficient mechanisms of quality control to balance cell survival and resistance to external and internal stress are required. Our objective was to test the relevance of cell quality control mechanisms for SCs maintenance, differentiation and resistance to cell death. We compared cell quality control in P19 stem cells (P19SCs) before and after differentiation (P19dCs). Differentiation of P19SCs resulted in alterations in parameters involved in cell survival and protein homeostasis, including the redox system, cardiolipin and lipid profiles, unfolded protein response, ubiquitin-proteasome and lysosomal systems, and signaling pathways controlling cell growth. In addition, P19SCs pluripotency was correlated with stronger antioxidant protection, modulation of apoptosis, and activation of macroautophagy, which all contributed to preserve SCs quality by increasing the threshold for cell death activation. Furthermore, our findings identify critical roles for the PI3K-AKT-MTOR pathway, as well as autophagic flux and apoptosis regulation in the maintenance of P19SCs pluripotency and differentiation potential. Abbreviations: 3-MA: 3-methyladenine; AKT/protein kinase B: thymoma viral proto-oncogene; AKT1: thymoma viral proto-oncogene 1; ATG: AuTophaGy-related; ATF6: activating transcription factor 6; BAX: BCL2-associated X protein; BBC3/PUMA: BCL2 binding component 3; BCL2: B cell leukemia/lymphoma 2; BNIP3L: BCL2/adenovirus E1B interacting protein 3-like; CASP3: caspase 3; CASP8: caspase 8; CASP9: caspase 9; CL: cardiolipin; CTSB: cathepsin B; CTSD: cathepsin D; DDIT3/CHOP: DNA-damage inducible transcript 3; DNM1L/DRP1: dynamin 1-like; DRAM1: DNA-damage regulated autophagy modulator 1; EIF2AK3/PERK: eukaryotic translation initiation factor 2 alpha kinase 3; EIF2S1/eIF2α: eukaryotic translation initiation factor 2, subunit alpha; ERN1/IRE1α: endoplasmic reticulum to nucleus signaling 1; ESCs: embryonic stem cells; KRT8/TROMA-1: cytokeratin 8; LAMP2A: lysosomal-associated membrane protein 2A; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MTOR: mechanistic target of rapamycin kinase; NANOG: Nanog homeobox; NAO: 10-N-nonyl acridine orange; NFE2L2/NRF2: nuclear factor, erythroid derived 2, like 2; OPA1: OPA1, mitochondrial dynamin like GTPase; P19dCs: P19 differentiated cells; P19SCs: P19 stem cells; POU5F1/OCT4: POU domain, class 5, transcription factor 1; PtdIns3K: phosphatidylinositol 3-kinase; RA: retinoic acid; ROS: reactive oxygen species; RPS6KB1/p70S6K: ribosomal protein S6 kinase, polypeptide 1; SCs: stem cells; SOD: superoxide dismutase; SHC1-1/p66SHC: src homology 2 domain-containing transforming protein C1, 66 kDa isoform; SOX2: SRY (sex determining region Y)-box 2; SQSTM1/p62: sequestosome 1; SPTAN1/αII-spectrin: spectrin alpha, non-erythrocytic 1; TOMM20: translocase of outer mitochondrial membrane 20; TRP53/p53: transformation related protein 53; TUBB3/betaIII-tubulin: tubulin, beta 3 class III; UPR: unfolded protein response; UPS: ubiquitin-proteasome system.

KW - Autophagy

KW - cell death

KW - metabolism

KW - redox system

KW - stem cells differentiation

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Cell quality control mechanisms maintain stemness and differentiation potential of P19 embryonic carcinoma cells

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