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Adora2b-elicited Per2 stabilization promotes a HIF-dependent metabolic switch crucial for myocardial adaptation to ischemia

Abstract

Adenosine signaling has been implicated in cardiac adaptation to limited oxygen availability. In a wide search for adenosine receptor A2b (Adora2b)-elicited cardioadaptive responses, we identified the circadian rhythm protein period 2 (Per2) as an Adora2b target. Adora2b signaling led to Per2 stabilization during myocardial ischemia, and in this setting, Per2−/− mice had larger infarct sizes compared to wild-type mice and loss of the cardioprotection conferred by ischemic preconditioning. Metabolic studies uncovered a limited ability of ischemic hearts in Per2−/− mice to use carbohydrates for oxygen-efficient glycolysis. This impairment was caused by a failure to stabilize hypoxia-inducible factor-1α (Hif-1α). Moreover, stabilization of Per2 in the heart by exposing mice to intense light resulted in the transcriptional induction of glycolytic enzymes and Per2-dependent cardioprotection from ischemia. Together, these studies identify adenosine-elicited stabilization of Per2 in the control of HIF-dependent cardiac metabolism and ischemia tolerance and implicate Per2 stabilization as a potential new strategy for treating myocardial ischemia.

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Figure 1: Consequences of adenosine signaling on Per2 induction.
Figure 2: Influence of transcriptional, translational and post-translational mechanisms on Per2 protein expression.
Figure 3: Functional role of Per2 during myocardial ischemia and ischemic preconditioning.
Figure 4: Consequences of Per2 deficiency on cardiac metabolism during myocardial ischemia and reperfusion.
Figure 5: Hif-1α functions as a link between adenosine-mediated Per2 signaling and metabolism.
Figure 6: Light-induced stabilization of cardiac Per2 expression provides potent protection from myocardial ischemia.

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Acknowledgements

The authors acknowledge S.A. Eltzschig (Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado Denver, Aurora, Colorado, USA) for artwork during manuscript preparation, A. Medway (Division of Cardiology, Department of Medicine, University of Colorado Denver, Aurora, Colorado, USA), K. Hoffmann, J. Macas and C. Zachskorn (Institute of Neurology (Edinger Institute), University of Frankfurt, Frankfurt, Germany) and M. Bonney (Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado Denver, Aurora, Colorado, USA) for technical assistance and C.-C. Lee (Department of Biochemistry and Molecular Biology, University of Texas Health Science Center, Houston, Texas, USA) for providing the Per1−/− mice. The present research work is supported by National Heart, Lung and Blood Institute grants R01-HL092188, R01-DK083385 and R01-HL098294 to H.K.E., R01-HL060569 to S.P.C. and 1K08HL102267 to T.E., Foundation for Anesthesia Education and Research grants to T.E. and H.K.E., an American Heart Association grant to T.E. and a Crohn's and Colitis Foundation of America grant to H.K.E.

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T.E. designed and supervised the study, wrote the manuscript and did mouse surgery. K.H. did western blots, RT-PCRs and siRNA knockdown studies. S.B. did western blots, coimmunoprecipitation, promoter studies, ELISAs and animal experiments. S.R. did western blots, RT-PCRs, ELISAs and mouse experiments. M.M. did immunohistochemistry and electron microscopy. L.A.W. isolated mouse myocytes and supervised the study. B.D.L. provided human heart samples. J.H., C.H.B. and D.J.K. did metabolic analysis. P.M.B., S.P.C. and H.K.E. supervised the study and wrote the manuscript.

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Correspondence to Tobias Eckle.

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Eckle, T., Hartmann, K., Bonney, S. et al. Adora2b-elicited Per2 stabilization promotes a HIF-dependent metabolic switch crucial for myocardial adaptation to ischemia. Nat Med 18, 774–782 (2012). https://doi.org/10.1038/nm.2728

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