title: Cooperation of adenosine with macrophage Toll-4 receptor agonists leads to increased glycolytic flux through the enhanced expression of PFKFB3 gene.
abstract: Macrophages activated through Toll receptor triggering increase the expression of the A(2A) and A(2B) adenosine receptors. In this study, we show that adenosine receptor activation enhances LPS-induced pfkfb3 expression, resulting in an increase of the key glycolytic allosteric regulator fructose 2,6-bisphosphate and the glycolytic flux. Using shRNA and differential expression of A(2A) and A(2B) receptors, we demonstrate that the A(2A) receptor mediates, in part, the induction of pfkfb3 by LPS, whereas the A(2B) receptor, with lower adenosine affinity, cooperates when high adenosine levels are present. pfkfb3 promoter sequence deletion analysis, site-directed mutagenesis, and inhibition by shRNAs demonstrated that HIF1α is a key transcription factor driving pfkfb3 expression following macrophage activation by LPS, whereas synergic induction of pfkfb3 expression observed with the A(2) receptor agonists seems to depend on Sp1 activity. Furthermore, levels of phospho-AMP kinase also increase, arguing for increased PFKFB3 activity by phosphorylation in long term LPS-activated macrophages. Taken together, our results show that, in macrophages, endogenously generated adenosine cooperates with bacterial components to increase PFKFB3 isozyme activity, resulting in greater fructose 2,6-bisphosphate accumulation. This process enhances the glycolytic flux and favors ATP generation helping to develop and maintain the long term defensive and reparative functions of the macrophages.
authors.affiliation: Facultad de Medicina, Centro Regional de Investigaciones Biomédicas, Albacete, Spain.
authors.forename: Almudena
authors.initials: A
authors.lastname: Ruiz-García
authors.forename: Eva
authors.initials: E
authors.lastname: Monsalve
authors.forename: Laura
authors.initials: L
authors.lastname: Novellasdemunt
authors.forename: Aurea
authors.initials: A
authors.lastname: Navarro-Sabaté
authors.forename: Anna
authors.initials: A
authors.lastname: Manzano
authors.forename: Samuel
authors.initials: S
authors.lastname: Rivero
authors.forename: Antonio
authors.initials: A
authors.lastname: Castrillo
authors.forename: Marta
authors.initials: M
authors.lastname: Casado
authors.forename: Jorge
authors.initials: J
authors.lastname: Laborda
authors.forename: Ramón
authors.initials: R
authors.lastname: Bartrons
authors.forename: María José M
authors.initials: MJ
authors.lastname: Díaz-Guerra
mesh_terms: D000241:Adenosine / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000096:biosynthesis / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D005635:Fructosediphosphates / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects* / Q000502:physiology; D006019:Glycolysis / Q000187:drug effects* / Q000502:physiology; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008262:Macrophage Activation / Q000187:drug effects / Q000502:physiology; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D018345:Mice, Knockout; D016297:Mutagenesis, Site-Directed; D025481:Phosphofructokinase-2 / Q000096:biosynthesis* / Q000235:genetics; D043705:Receptor, Adenosine A2A / Q000235:genetics / Q000378:metabolism; D043704:Receptor, Adenosine A2B; D017384:Sequence Deletion; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism
chemical_list: D005635:Fructosediphosphates; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D007527:Isoenzymes; D008070:Lipopolysaccharides; D043705:Receptor, Adenosine A2A; D043704:Receptor, Adenosine A2B; D016329:Sp1 Transcription Factor; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; C027652:fructose 2,6-diphosphate; D000255:Adenosine Triphosphate; D025481:Phosphofructokinase-2; D000241:Adenosine