AICAR Stimulates the Pluripotency Transcriptional Complex in Embryonic Stem Cells Mediated by PI3K, GSK3ß, and ß-Catenin.
Por:
Alba G, Martínez R, Postigo-Corrales F, López S, Santa-María C, Jiménez J, Cahuana GM, Soria B, Bedoya FJ and Tejedo JR
Publicada:
18 ago 2020
Ahead of Print:
4 ago 2020
Resumen:
Pluripotent stem cells maintain the property of self-renewal and differentiate into all cell types under clear environments. Though the gene regulatory mechanism for pluripotency has been investigated in recent years, it is still not completely understood. Here, we show several signaling pathways involved in the maintenance of pluripotency. To investigate whether AMPK is involved in maintaining the pluripotency in mouse embryonic stem cells (mESCs) and elucidating the possible molecular mechanisms, implicated D3 and R1/E mESC lines were used in this study. Cells were cultured in the absence or presence of LIF and treated with 1 mM and 0.5 mM 5-aminoimidazole-4-carboxamide-1-ß-d-ribofuranoside (AICAR), 2 mM metformin, compound C, and the PI3K inhibitor LY294002 for 24, 72, and 120 h. The levels of Nanog, Oct3/4, and REX1 and Brachyury, Notch2, and Gata4 mRNAs and Nanog or OCT3/4 protein levels were analyzed. Alkaline phosphatase and the cellular cycle were determined. The pGSK3ß, GSK3ß, p-ß-catenin, and ß-catenin protein levels were also investigated. We found that AMPK activators such as AICAR and metformin increase mRNA expression of pluripotency markers and decrease mRNA expression of differentiation markers in R1/E and D3 ES cells. AICAR increases phosphatase activity and arrests the cellular cycle in the G1 phase in these cells. We describe that AICAR effects were mediated by AMPK activation using a chemical inhibitor or by silencing this gene. AICAR effects were also mediated by PI3K, GSK3ß, and ß-catenin in R1/E ES cells. According to our findings, we provide a mechanism by which AICAR increases and maintains a pluripotency state through enhanced Nanog expression, involving AMPK/PI3K and p-GSK3ß Ser21/9 pathways backing up the AICAR function as a potential target for this drug controlling pluripotency. The highlights of this study are that AICAR (5-aminoimidazole-4-carboxamied-1-b-riboside), an AMP protein kinase (AMPK) activator, blocks the ESC differentiation and AMPK is a key enzyme for pluripotency and shows valuable data to clarify the molecular pluripotency mechanism.
Filiaciones:
Alba G:
Department of Medical Biochemistry and Molecular Biology, Universidad de Sevilla, Seville 41009, Spain
Martínez R:
Department of Regeneration and Cell Therapy, Andalusian Center for Molecular Biology and Regenerative Medicine-CABIMER, Universidad Pablo de Olavide-University of Seville-CSIC, Seville 41013, Spain
Postigo-Corrales F:
Department of Regeneration and Cell Therapy, Andalusian Center for Molecular Biology and Regenerative Medicine-CABIMER, Universidad Pablo de Olavide-University of Seville-CSIC, Seville 41013, Spain
López S:
Department of Medical Biochemistry and Molecular Biology, Universidad de Sevilla, Seville 41009, Spain
Santa-María C:
Department of Biochemistry and Molecular Biology, Universidad de Sevilla, Seville 41009, Spain
Jiménez J:
Department of Medical Biochemistry and Molecular Biology, Universidad de Sevilla, Seville 41009, Spain
Cahuana GM:
Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville 41013, Spain
Biomedical Research Network for Diabetes and Related Metabolic Diseases-CIBERDEM, Instituto de Salud Carlos III, Madrid 28029, Spain
:
Department of Regeneration and Cell Therapy, Andalusian Center for Molecular Biology and Regenerative Medicine-CABIMER, Universidad Pablo de Olavide-University of Seville-CSIC, Seville 41013, Spain
Biomedical Research Network for Diabetes and Related Metabolic Diseases-CIBERDEM, Instituto de Salud Carlos III, Madrid 28029, Spain
Cell Therapy Network, Madrid (RED-TERCEL), Instituto de Salud Carlos III, Madrid 28029, Spain
Universidad Miguel Hernández, Alicante 03550, Spain
Bedoya FJ:
Department of Regeneration and Cell Therapy, Andalusian Center for Molecular Biology and Regenerative Medicine-CABIMER, Universidad Pablo de Olavide-University of Seville-CSIC, Seville 41013, Spain
Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville 41013, Spain
Biomedical Research Network for Diabetes and Related Metabolic Diseases-CIBERDEM, Instituto de Salud Carlos III, Madrid 28029, Spain
Cell Therapy Network, Madrid (RED-TERCEL), Instituto de Salud Carlos III, Madrid 28029, Spain
Tejedo JR:
Department of Regeneration and Cell Therapy, Andalusian Center for Molecular Biology and Regenerative Medicine-CABIMER, Universidad Pablo de Olavide-University of Seville-CSIC, Seville 41013, Spain
Department of Molecular Biology and Biochemical Engineering, Universidad Pablo de Olavide, Seville 41013, Spain
Biomedical Research Network for Diabetes and Related Metabolic Diseases-CIBERDEM, Instituto de Salud Carlos III, Madrid 28029, Spain
Cell Therapy Network, Madrid (RED-TERCEL), Instituto de Salud Carlos III, Madrid 28029, Spain
Open Access
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