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PI3K–Akt signaling controls PFKFB3 expression during human T-lymphocyte activation

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Abstract

Lymphocyte activation is associated with rapid increase of both the glycolytic activator fructose 2,6-bisphosphate (Fru-2,6-P2) and the enzyme responsible for its synthesis, 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (PFK-2/FBPase-2). PFKFB3 gene, which encodes for the most abundant PFK-2 isoenzyme in proliferating tissues, has been found overexpressed during cell activation in several models, including immune cells. However, there is limited knowledge on the pathways underlying PFKFB3 regulation in human T-lymphocytes, and the role of this gene in human immune response. The aim of this work is to elucidate the molecular mechanisms of PFKFB3 induction during human T-lymphocyte activation by mitotic agents. The results obtained showed PFKFB3 induction during human T-lymphocyte activation by mitogens such as phytohemagglutinin (PHA). PFKFB3 increase occurred concomitantly with GLUT-1, HK-II, and PCNA upregulation, showing that mitotic agents induce a metabolic reprograming process that is required for T-cell proliferation. PI3K–Akt pathway inhibitors, Akti-1/2 and LY294002, reduced PFKFB3 gene induction by PHA, as well as Fru-2,6-P2 and lactate production. Moreover, both inhibitors blocked activation and proliferation in response to PHA, showing the importance of PI3K/Akt signaling pathway in the antigen response of T-lymphocytes. These results provide a link between metabolism and T-cell antigen receptor signaling in human lymphocyte biology that can help to better understand the importance of modulating both pathways to target complex diseases involving the activation of the immune system.

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Abbreviations

3PO:

3-(3-pyridinyl)-1-(4-pyridinyl)-2-propen-1-one

7-AAD:

7-Aminoactinomycin D

AICD:

Activation-induced cell death

CFSE:

Carboxyfluorescein succinimidyl ester

ConA:

Concanavalin A

FBS:

Fetal bovine serum

Fru-2,6-P2 :

Fructose 2,6-bisphosphate

GLUT-1:

Glucose transporter 1

HIF:

Hypoxia-inducible factor

HK-II:

Hexokinase-II

IL2RA:

Interleukin-2 receptor alpha chain

LPS:

Lipopolysaccharide

MFI:

Mean fluorescence intensity

mTORC1:

Mammalian target of rapamycin

mTORC1:

Mammalian target of rapamycin complex 1

OXPHOS:

Oxidative phosphorylation

PBMCs:

Peripheral blood mononuclear cells

PCNA:

Proliferating cell nuclear antigen

PFK-1:

Phosphofructokinase-1

PFK-2/FBPase-2:

6-Phosphofructo-2-Kinase/Fructose-2,6-Bisphosphatase

PFKFB3:

6-Phosphofructo-2-Kinase/Fructose-2,6-Bisphosphatase 3

PHA:

Phytohemagglutinin

P-S6:

S6 ribosomal protein

TCR:

T-cell antigen receptor

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Acknowledgements

We are grateful to B. Torrejón from the Microscopy Unit of Centres Científics i Tecnològics of the Universitat de Barcelona (CCiT-UB) for excellent technical assistance and advice; A. M. Cosialls for technical advice in blood cells cultures; J. L. Rosa and F. Viñals for providing reagents; P. Giménez-Bonafé for English revision and E. Adanero for technical assistance.

Funding

This work was supported by Instituto de Salud Carlos III—FIS [PI13/0096] and FIS [PI17/00412]—and Fondo Europeo de Desarrollo Regional (FEDER), and by Astellas European Foundation Award (13th European Society of Transplantation). HS was recipient of a fellowship from Secretaria d’Universitats i Recerca del Departament d’Economia i Coneixement de la Generalitat de Catalunya and AR was recipient of a fellowship from the University of Barcelona.

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    Authors and Affiliations

    1. Departament de Ciències Fisiològiques, Facultat de Medicina i Ciències de la Salut, Universitat de Barcelona, C/Feixa Llarga, s/n 08907 L’Hospitalet de Llobregat, Barcelona, Spain

      Helga Simon-Molas, Claudia Arnedo-Pac, Ana Rodríguez-García, Àurea Navarro-Sabaté, Anna Manzano & Ramon Bartrons

    2. Departament de Nefrologia, Hospital Universitari de Bellvitge, IDIBELL, Barcelona, Spain

      Pere Fontova, Anna Vidal-Alabró & Núria Lloberas

    3. Centres Científics i Tecnològics, Universitat de Barcelona, Barcelona, Spain

      Esther Castaño

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    1. Helga Simon-Molas
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    Contributions

    RB and AM conceived and designed the experiments; HS, CA, PF, and AV performed the experiments; HS, CA, AM, PF, AV, and EC analyzed the data; NLL, AR, and AN critically commented and revised the work; and HS, RB, and AM wrote the manuscript.

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    Correspondence to Ramon Bartrons.

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    Helga Simon-Molas and Claudia Arnedo-Pac have contributed equally to this work.

    Anna Manzano and Ramon Bartrons share senior co-authorship.

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    Simon-Molas, H., Arnedo-Pac, C., Fontova, P. et al. PI3K–Akt signaling controls PFKFB3 expression during human T-lymphocyte activation. Mol Cell Biochem 448, 187–197 (2018). https://doi.org/10.1007/s11010-018-3325-9

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