Crosstalk between the equilibrative nucleoside transporter ENT2 and alveolar Adora2b adenosine receptors dampens acute lung injury
Tobias Eckle
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorKelly Hughes
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorHeidi Ehrentraut
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorKelley S. Brodsky
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorPeter Rosenberger
Department of Anesthesiology and Intensive Care Medicine, University Hospital, Tübingen, Germany
Search for more papers by this authorDoo-Sup Choi
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota, USA
Search for more papers by this authorKatya Ravid
Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts, USA
Search for more papers by this authorTingting Weng
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorYang Xia
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorMichael R. Blackburn
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorCorresponding Author
Holger K. Eltzschig
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Correspondence: Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, 12700 E. 19th Ave., Mailstop B112, Research Complex 2, Room 7124, Aurora, CO 80045, USA. E-mail: [email protected]Search for more papers by this authorTobias Eckle
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorKelly Hughes
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorHeidi Ehrentraut
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorKelley S. Brodsky
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Search for more papers by this authorPeter Rosenberger
Department of Anesthesiology and Intensive Care Medicine, University Hospital, Tübingen, Germany
Search for more papers by this authorDoo-Sup Choi
Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota, USA
Search for more papers by this authorKatya Ravid
Department of Medicine, Boston University School of Medicine, Boston, Massachusetts, USA
Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts, USA
Search for more papers by this authorTingting Weng
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorYang Xia
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorMichael R. Blackburn
Department of Biochemistry and Molecular Biology, The University of Texas Medical School at Houston, Houston, Texas, USA
Search for more papers by this authorCorresponding Author
Holger K. Eltzschig
Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, Aurora, Colorado, USA
Correspondence: Mucosal Inflammation Program, Department of Anesthesiology, University of Colorado School of Medicine, 12700 E. 19th Ave., Mailstop B112, Research Complex 2, Room 7124, Aurora, CO 80045, USA. E-mail: [email protected]Search for more papers by this authorAbstract
The signaling molecule adenosine has been implicated in attenuating acute lung injury (ALI). Adenosine signaling is terminated by its uptake through equilibrative nucleoside transporters (ENTs). We hypothesized that ENT-dependent adenosine uptake could be targeted to enhance adenosine-mediated lung protection. To address this hypothesis, we exposed mice to high-pressure mechanical ventilation to induce ALI. Initial studies demonstrated time-dependent repression of ENT1 and ENT2 transcript and protein levels during ALI. To examine the contention that ENT repression represents an endogenous adaptive response, we performed functional studies with the ENT inhibitor dipyridamole. Dipyridamole treatment (1 mg/kg; EC50=10 μM) was associated with significant increases in ALI survival time (277 vs. 395 min; P<0.05). Subsequent studies in gene-targeted mice for Ent1 or Ent2 revealed a selective phenotype in Ent2-/- mice, including attenuated pulmonary edema and improved gas exchange during ALI in conjunction with elevated adenosine levels in the bronchoalveolar fluid. Furthermore, studies in genetic models for adenosine receptors implicated the A2B adenosine receptor (Adora2b) in mediating ENT-dependent lung protection. Notably, dipyridamole-dependent attenuation of lung inflammation was abolished in mice with alveolar epithelial Adora2b gene deletion. Our newly identified crosstalk pathway between ENT2 and alveolar epithelial Adora2b in lung protection during ALI opens possibilities for combined therapies targeted to this protein set.—Eckle, T., Hughes, K., Ehrentraut, H., Brodsky, K. S., Rosenberger, P., Choi, D.-S., Ravid, K., Weng, T., Xia, Y., Blackburn, M. R.Eltzschig, H. K., Crosstalk between the equilibrative nucleoside transporter ENT2 and alveolar Adora2b adenosine receptors dampens acute lung injury. FASEB J. 27, 3078–3089 (2013). www.fasebj.org
REFERENCES
- 1Ranieri, V. M., Rubenfeld, G. D., Thompson, B. T., Ferguson, N. D., Caldwell, E., Fan, E., Camporota, L., and Slutsky, A. S. (2012) Acute respiratory distress syndrome: the Berlin definition. JAMA 307, 2526–2533
- 2Ware, L. B., and Matthay, M. A. (2000) The acute respiratory distress syndrome. N. Engl. J. Med. 342, 1334–1349
- 3Eckle, T., Koeppen, M., and Eltzschig, H. K. (2009) Role of extracellular adenosine in acute lung injury. Physiology (Bethesda) 24, 298–306
- 4Eltzschig, H. K., Sitkovsky, M. V., and Robson, S. C. (2012) Purinergic signaling during inflammation. N. Engl. J. Med. 367, 2322–2333
- 5Rubenfeld, G. D., Caldwell, E., Peabody, E., Weaver, J., Martin, D. P., Neff, M., Stern, E. J., and Hudson, L. D. (2005) Incidence and outcomes of acute lung injury. N. Engl. J. Med. 353, 1685–1693
- 6Herridge, M. S., Tansey, C. M., Matte, A., Tomlinson, G., Diaz-Granados, N., Cooper, A., Guest, C. B., Mazer, C. D., Mehta, S., Stewart, T. E., Kudlow, P., Cook, D., Slutsky, A. S., and Cheung, A. M. (2011) Functional disability 5 years after acute respiratory distress syndrome. N. Engl. J. Med. 364, 1293–1304
- 7Eltzschig, H. K., and Carmeliet, P. (2011) Hypoxia and inflammation. N. Engl. J. Med. 364, 656–665
- 8Thiel, M., Chouker, A., Ohta, A., Jackson, E., Caldwell, C., Smith, P., Lukashev, D., Bittmann, I., and Sitkovsky, M. V. (2005) Oxygenation inhibits the physiological tissue-protecting mechanism and thereby exacerbates acute inflammatory lung injury. PLoS Biol. 3, e174
- 9Martin, T. R. (2002) Neutrophils and lung injury: getting it right. J. Clin. Invest. 110, 1603–1605
- 10Eckle, T., Fullbier, L., Grenz, A., and Eltzschig, H. K. (2008) Usefulness of pressure-controlled ventilation at high inspiratory pressures to induce acute lung injury in mice. Am. J. Physiol. Lung Cell. Mol. Physiol. 295, L718–L724
- 11Eckle, T., Fullbier, L., Wehrmann, M., Khoury, J., Mittelbronn, M., Ibla, J., Rosenberger, P., and Eltzschig, H. K. (2007) Identification of ectonucleotidases CD39 and CD73 in innate protection during acute lung injury. J. Immunol. 178, 8127–8137
- 12Reutershan, J., Vollmer, I., Stark, S., Wagner, R., Ngamsri, K. C., and Eltzschig, H. K. (2008) Adenosine and inflammation: CD39 and CD73 are critical mediators in LPS-induced PMN trafficking into the lungs. FASEB J. 23, 473–482
- 13Van Linden, A., and Eltzschig, H. K. (2007) Role of pulmonary adenosine during hypoxia: extracellular generation, signaling and metabolism by surface adenosine deaminase/CD26. Expert Opin. Biol. Ther. 7, 1437–1447
- 14Eckle, T., Grenz, A., Laucher, S., and Eltzschig, H. K. (2008) A2B adenosine receptor signaling attenuates acute lung injury by enhancing alveolar fluid clearance in mice. J. Clin. Invest. 118, 3301–3315
- 15Zhou, Y., Schneider, D. J., Morschl, E., Song, L., Pedroza, M., Karmouty-Quintana, H., Le, T., Sun, C. X., and Blackburn, M. R. (2011) Distinct roles for the A2B adenosine receptor in acute and chronic stages of bleomycin-induced lung injury. J. Immunol. 186, 1097–1106
- 16Lu, Q., Harrington, E. O., Newton, J., Casserly, B., Radin, G., Warburton, R., Zhou, Y., Blackburn, M. R., and Rounds, S. (2010) Adenosine protected against pulmonary edema through transporter- and receptor A2-mediated endothelial barrier enhancement. Am. J. Physiol. Lung Cell. Mol. Physiol. 298, L755–L767
- 17Zhou, Y., Mohsenin, A., Morschl, E., Young, H. W., Molina, J. G., Ma, W., Sun, C. X., Martinez-Valdez, H., and Blackburn, M. R. (2009) Enhanced airway inflammation and remodeling in adenosine deaminase-deficient mice lacking the A2B adenosine receptor. J. Immunol. 182, 8037–8046
- 18Eltzschig, H. K., and Collard, C. D. (2004) Vascular ischaemia and reperfusion injury. Br. Med. Bull. 70, 71–86
- 19Schingnitz, U., Hartmann, K., Macmanus, C. F., Eckle, T., Zug, S., Colgan, S. P., and Eltzschig, H. K. (2010) Signaling through the A2B adenosine receptor dampens endotoxin-induced acute lung injury. J. Immunol. 184, 5271–5279
- 20Eltzschig, H. K., and Eckle, T. (2011) Ischemia and reperfusion—from mechanism to translation. Nat. Med. 17, 1391–1401
- 21Hart, M. L., Grenz, A., Gorzolla, I. C., Schittenhelm, J., Dalton, J. H., and Eltzschig, H. K. (2011) Hypoxia-inducible factor-1α-dependent protection from intestinal ischemia/reperfusion injury involves ecto-5′-nucleotidase (CD73) and the A2B adenosine receptor. J. Immunol. 186, 4367–4374
- 22Hart, M. L., Gorzolla, I. C., Schittenhelm, J., Robson, S. C., and Eltzschig, H. K. (2010) SP1-dependent induction of CD39 facilitates hepatic ischemic preconditioning. J. Immunol. 184, 4017–4024
- 23Hart, M. L., Jacobi, B., Schittenhelm, J., Henn, M., and Eltzschig, H. K. (2009) Cutting Edge: A2B Adenosine receptor signaling provides potent protection during intestinal ischemia/reperfusion injury. J. Immunol. 182, 3965–3968
- 24Hart, M. L., Henn, M., Kohler, D., Kloor, D., Mittelbronn, M., Gorzolla, I. C., Stahl, G. L., and Eltzschig, H. K. (2008) Role of extracellular nucleotide phosphohydrolysis in intestinal ischemia-reperfusion injury. FASEB J. 22, 2784–2797
- 25Hart, M. L., Much, C., Gorzolla, I. C., Schittenhelm, J., Kloor, D., Stahl, G. L., and Eltzschig, H. K. (2008) Extracellular adenosine production by ecto-5′-nucleotidase protects during murine hepatic ischemic preconditioning. Gastroenterology 135, 1739.e3–1750.e3
- 26Hart, M. L., Kohler, D., Eckle, T., Kloor, D., Stahl, G. L., and Eltzschig, H. K. (2008) Direct treatment of mouse or human blood with soluble 5′-nucleotidase inhibits platelet aggregation. Arterioscler. Thromb. Vasc. Biol. 28, 1477–1483
- 27Eltzschig, H. K., Macmanus, C. F., and Colgan, S. P. (2008) Neutrophils as sources of extracellular nucleotides: functional consequences at the vascular interface. Trends Cardiovasc. Med. 18, 103–107
- 28Faigle, M., Seessle, J., Zug, S., El Kasmi, K. C., and Eltzschig, H. K. (2008) ATP release from vascular endothelia occurs across Cx43 hemichannels and is attenuated during hypoxia. PLoS One 3, e2801
- 29Eltzschig, H. K., Weissmuller, T., Mager, A., and Eckle, T. (2006) Nucleotide metabolism and cell-cell interactions. Methods Mol. Biol. 341, 73–87
- 30Eltzschig, H. K., Eckle, T., Mager, A., Kuper, N., Karcher, C., Weissmuller, T., Boengler, K., Schulz, R., Robson, S. C., and Colgan, S. P. (2006) ATP release from activated neutrophils occurs via connexin 43 and modulates adenosine-dependent endothelial cell function. Circ. Res. 99, 1100–1108
- 31Eltzschig, H. K., Ibla, J. C., Furuta, G. T., Leonard, M. O., Jacobson, K. A., Enjyoji, K., Robson, S. C., and Colgan, S. P. (2003) Coordinated adenine nucleotide phosphohydrolysis and nucleoside signaling in posthypoxic endothelium: role of ectonucleotidases and adenosine A2B receptors. J. Exp. Med. 198, 783–796
- 32Eltzschig, H. K. (2009) Adenosine: an old drug newly discovered. Anesthesiology 111, 904–915
- 33Tsukamoto, H., Chernogorova, P., Ayata, K., Gerlach, U. V., Rughani, A., Ritchey, J. W., Ganesan, J., Follo, M., Zeiser, R., Thompson, L. F., and Idzko, M. Deficiency of CD73/ecto-5′-nucleotidase in mice enhances acute graft-versus-host disease. Blood 119, 4554–4564
- 34Grenz, A., Bauerle, J. D., Dalton, J. H., Ridyard, D., Badulak, A., Tak, E., McNamee, E. N., Clambey, E., Moldovan, R., Reyes, G., Klawitter, J., Ambler, K., Magee, K., Christians, U., Brodsky, K. S., Ravid, K., Choi, D. S., Wen, J., Lukashev, D., Blackburn, M. R., Osswald, H., Coe, I. R., Nurnberg, B., Haase, V. H., Xia, Y., Sitkovsky, M., and Eltzschig, H. K. (2012) Equilibrative nucleoside transporter 1 (ENT1) regulates postischemic blood flow during acute kidney injury in mice. J. Clin. Invest. 122, 693–710
- 35Morote-Garcia, J. C., Rosenberger, P., Nivillac, N. M., Coe, I. R., and Eltzschig, H. K. (2009) Hypoxia-inducible factor-dependent repression of equilibrative nucleoside transporter 2 attenuates mucosal inflammation during intestinal hypoxia. Gastroenterology 136, 607–618
- 36Loffler, M., Morote-Garcia, J. C., Eltzschig, S. A., Coe, I. R., and Eltzschig, H. K. (2007) Physiological roles of vascular nucleoside transporters. Arterioscler. Thromb. Vasc. Biol. 27, 1004–1013
- 37Eltzschig, H. K., Abdulla, P., Hoffman, E., Hamilton, K. E., Daniels, D., Schonfeld, C., Loffler, M., Reyes, G., Duszenko, M., Karhausen, J., Robinson, A., Westerman, K. A., Coe, I. R., and Colgan, S. P. (2005) HIF-1-dependent repression of equilibrative nucleoside transporter (ENT) in hypoxia. J. Exp. Med. 202, 1493–1505
- 38Alva, J. A., Zovein, A. C., Monvoisin, A., Murphy, T., Salazar, A., Harvey, N. L., Carmeliet, P., and Iruela-Arispe, M. L. (2006) VE-Cadherin-Cre-recombinase transgenic mouse: a tool for lineage analysis and gene deletion in endothelial cells. Dev. Dyn. 235, 759–767
- 39Schwenk, F., Baron, U., and Rajewsky, K. (1995) A cre-transgenic mouse strain for the ubiquitous deletion of loxP-flanked gene segments including deletion in germ cells. Nucleic Acids Res. 23, 5080–5081
- 40Okubo, T., and Hogan, B. L. (2004) Hyperactive Wnt signaling changes the developmental potential of embryonic lung endoderm. J. Biol. 3, 11
- 41Ko, K. R., Ngai, A. C., and Winn, H. R. (1990) Role of adenosine in regulation of regional cerebral blood flow in sensory cortex. Am. J. Physiol. 259, H1703–H1708
- 42Kim, H. H., Sawada, N., Soydan, G., Lee, H. S., Zhou, Z., Hwang, S. K., Waeber, C., Moskowitz, M. A., and Liao, J. K. (2008) Additive effects of statin and dipyridamole on cerebral blood flow and stroke protection. J. Cereb. Blood Flow Metab. 28, 1285–1293
- 43Ward, J. L., Sherali, A., Mo, Z. P., and Tse, C. M. (2000) Kinetic and pharmacological properties of cloned human equilibrative nucleoside transporters, ENT1 and ENT2, stably expressed in nucleoside transporter-deficient PK15 cells. Ent2 exhibits a low affinity for guanosine and cytidine but a high affinity for inosine. J. Biol. Chem. 275, 8375–8381
- 44Wakamiya, M., Blackburn, M. R., Jurecic, R., McArthur, M. J., Geske, R. S., Cartwright, J., Jr., Mitani, K., Vaishnav, S., Belmont, J. W., Kellems, R. E., Finegoldt, M. J., Montgomery, C. A. Jr., Bradley, A., and Caskey, C. T. (1995) Disruption of the adenosine deaminase gene causes hepatocellular impairment and perinatal lethality in mice. Proc. Natl. Acad. Sci. U. S. A. 92, 3673–3677
- 45Blackburn, M. R., Volmer, J. B., Thrasher, J. L., Zhong, H., Crosby, J. R., Lee, J. J., and Kellems, R. E. (2000) Metabolic consequences of adenosine deaminase deficiency in mice are associated with defects in alveogenesis, pulmonary inflammation, and airway obstruction. J. Exp. Med. 192, 159–170
- 46Yang, D., Zhang, Y., Nguyen, H. G., Koupenova, M., Chauhan, A. K., Makitalo, M., Jones, M. R., St Hilaire, C., Seldin, D. C., Toselli, P., Lamperti, E., Schreiber, B. M., Gavras, H., Wagner, D. D., and Ravid, K. (2006) The A2B adenosine receptor protects against inflammation and excessive vascular adhesion. J. Clin. Invest. 116, 1913–1923
- 47Lahm, A., Uhl, M., Lehr, H. A., Ihling, C., Kreuz, P. C., and Haberstroh, J. (2004) Photoshop-based image analysis of canine articular cartilage after subchondral damage. Arch. Orthop. Trauma Surg. 124, 431–436
- 48Lehr, H. A., Mankoff, D. A., Corwin, D., Santeusanio, G., and Gown, A. M. (1997) Application of Photoshop-based image analysis to quantification of hormone receptor expression in breast cancer. J. Histochem. Cytochem. 45, 1559–1565
- 49Sitkovsky, M., and Lukashev, D. (2005) Regulation of immune cells by local-tissue oxygen tension: HIF1α and adenosine receptors. Nat. Rev. Immunol. 5, 712–721
- 50Sitkovsky, M. V., Lukashev, D., Apasov, S., Kojima, H., Koshiba, M., Caldwell, C., Ohta, A., and Thiel, M. (2004) Physiological control of immune response and inflammatory tissue damage by hypoxia-inducible factors and adenosine A2A receptors. Annu. Rev. Immunol. 22, 657–682
- 51Ohta, A., and Sitkovsky, M. (2001) Role of G-protein-coupled adenosine receptors in downregulation of inflammation and protection from tissue damage. Nature 414, 916–920
- 52Koeppen, M., Eckle, T., and Eltzschig, H. K. (2011) Pressure controlled ventilation to induce acute lung injury in mice. J. Vis. Exp. 51, 2525
- 53Koeppen, M., McNamee, E. N., Brodsky, K. S., Aherne, C. M., Faigle, M., Downey, G. P., Colgan, S. P., Evans, C. M., Schwartz, D. A., and Eltzschig, H. K. (2012) Detrimental role of the airway mucin Muc5ac during ventilator-induced lung injury. [E-pub ahead of print] Mucosal Immunol. doi:10.1038/mi.2012.114
10.1038/mi.2012.114 Google Scholar
- 54Choi, D. S., Cascini, M. G., Mailliard, W., Young, H., Paredes, P., McMahon, T., Diamond, I., Bonci, A., and Messing, R. O. (2004) The type 1 equilibrative nucleoside transporter regulates ethanol intoxication and preference. Nat. Neurosci. 7, 855–861
- 55Cagnina, R. E., Ramos, S. I., Marshall, M. A., Wang, G., Frazier, C. R., and Linden, J. (2009) Adenosine A2B receptors are highly expressed on murine type II alveolar epithelial cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 297, L467–L474
10.1152/ajplung.90553.2008 Google Scholar
- 56Grenz, A., Osswald, H., Eckle, T., Yang, D., Zhang, H., Tran, Z. V., Klingel, K., Ravid, K., and Eltzschig, H. K. (2008) The reno-vascular A2B adenosine receptor protects the kidney from ischemia. PLoS Med. 5, e137
- 57Eckle, T., Faigle, M., Grenz, A., Laucher, S., Thompson, L. F., and Eltzschig, H. K. (2008) A2B adenosine receptor dampens hypoxia-induced vascular leak. Blood 111, 2024–2035
- 58Eckle, T., Krahn, T., Grenz, A., Kohler, D., Mittelbronn, M., Ledent, C., Jacobson, M. A., Osswald, H., Thompson, L. F., Unertl, K., and Eltzschig, H. K. (2007) Cardioprotection by ecto-5′-nucleotidase (CD73) and A2B adenosine receptors. Circulation 115, 1581–1590
- 59Oliver, K. M., Lenihan, C. R., Bruning, U., Cheong, A., Laffey, J. G., McLoughlin, P., Taylor, C. T., and Cummins, E. P. (2012) Hypercapnia induces cleavage and nuclear localization of RelB protein, giving insight into CO2 sensing and signaling. J. Biol. Chem. 287, 14004–14011
- 60Taylor, C. T., and Cummins, E. P. (2011) Regulation of gene expression by carbon dioxide. J. Physiol. 589, 797–803
- 61Casanello, P., Torres, A., Sanhueza, F., Gonzalez, M., Farias, M., Gallardo, V., Pastor-Anglada, M., San Martin, R., and Sobrevia, L. (2005) Equilibrative nucleoside transporter 1 expression is downregulated by hypoxia in human umbilical vein endothelium. Circ. Res. 97, 16–24
- 62Chaudary, N., Naydenova, Z., Shuralyova, I., and Coe, I. R. (2004) Hypoxia regulates the adenosine transporter, mENT1, in the murine cardiomyocyte cell line, HL-1. Cardiovasc. Res. 61, 780–788
- 63Taylor, C. T., and McElwain, J. C. (2010) Ancient atmospheres and the evolution of oxygen sensing via the hypoxia-inducible factor in metazoans. Physiology (Bethesda) 25, 272–279
- 64Cummins, E. P., and Taylor, C. T. (2005) Hypoxia-responsive transcription factors. Pflügers Arch. 450, 363–371
- 65Cummins, E. P., Berra, E., Comerford, K. M., Ginouves, A., Fitzgerald, K. T., Seeballuck, F., Godson, C., Nielsen, J. E., Moynagh, P., Pouyssegur, J., and Taylor, C. T. (2006) Prolyl hydroxylase-1 negatively regulates IkB kinase-β, giving insight into hypoxia-induced NFκB activity. Proc. Natl. Acad. Sci. U. S. A. 103, 18154–18159
- 66Taylor, C. T. (2008) Interdependent roles for hypoxia inducible factor and nuclear factor-κB in hypoxic inflammation. J. Physiol. 586, 4055–4059
- 67Cummins, E. P., Seeballuck, F., Keely, S. J., Mangan, N. E., Callanan, J. J., Fallon, P. G., and Taylor, C. T. (2008) The hydroxylase inhibitor dimethyloxalylglycine is protective in a murine model of colitis. Gastroenterology 134, 156–165
- 68Aherne, C. M., Collins, C. B., Masterson, J. C., Tizzano, M., Boyle, T. A., Westrich, J. A., Parnes, J. A., Furuta, G. T., Rivera-Nieves, J., and Eltzschig, H. K. (2012) Neuronal guidance molecule netrin-1 attenuates inflammatory cell trafficking during acute experimental colitis. Gut 61, 695–705
- 69Grenz, A., Dalton, J. H., Bauerle, J. D., Badulak, A., Ridyard, D., Gandjeva, A., Aherne, C. M., Brodsky, K. S., Kim, J. H., Tuder, R. M., and Eltzschig, H. K. (2011) Partial netrin-1 deficiency aggravates acute kidney injury. PLoS One 6, e14812
- 70Rosenberger, P., Schwab, J. M., Mirakaj, V., Masekowsky, E., Mager, A., Morote-Garcia, J. C., Unertl, K., and Eltzschig, H. K. (2009) Hypoxia-inducible factor-dependent induction of netrin-1 dampens inflammation caused by hypoxia. Nat. Immunol. 10, 195–202
- 71Eckle, T., Hartmann, K., Bonney, S., Reithel, S., Mittelbronn, M., Walker, L. A., Lowes, B. D., Han, J., Borchers, C. H., Buttrick, P. M., Kominsky, D. J., Colgan, S. P., and Eltzschig, H. K. (2012) Adora2b-elicited Per2 stabilization promotes a HIF-dependent metabolic switch crucial for myocardial adaptation to ischemia. Nat. Med. 18, 774–782
- 72Koeppen, M., Harter, P. N., Bonney, S., Bonney, M., Reithel, S., Zachskorn, C., Mittelbronn, M., and Eckle, T. (2012) Adora2b signaling on bone marrow derived cells dampens myocardial ischemia-reperfusion injury. Anesthesiology 116, 1245–1257
- 73Eckle, T., Kohler, D., Lehmann, R., El Kasmi, K. C., and Eltzschig, H. K. (2008) Hypoxia-inducible factor-1 is central to cardioprotection: a new paradigm for ischemic preconditioning. Circulation 118, 166–175
- 74Dixon, B. S., Beck, G. J., Vazquez, M. A., Greenberg, A., Delmez, J. A., Allon, M., Dember, L. M., Himmelfarb, J., Gassman, J. J., Greene, T., Radeva, M. K., Davidson, I. J., Ikizler, T. A., Braden, G. L., Fenves, A. Z., Kaufman, J. S., Cotton, J. R., Jr., Martin, K. J., McNeil, J. W., Rahman, A., Lawson, J. H., Whiting, J. F., Hu, B., Meyers, C. M., Kusek, J. W., and Feldman, H. I. (2009) Effect of dipyridamole plus aspirin on hemodialysis graft patency. N. Engl. J. Med. 360, 2191–2201
- 75Sacco, R. L., Diener, H. C., Yusuf, S., Cotton, D., Ounpuu, S., Lawton, W. A., Palesch, Y., Martin, R. H., Albers, G. W., Bath, P., Bornstein, N., Chan, B. P., Chen, S. T., Cunha, L., Dahlof, B., De Keyser, J., Donnan, G. A., Estol, C., Gorelick, P., Gu, V., Hermansson, K., Hilbrich, L., Kaste, M., Lu, C., Machnig, T., Pais, P., Roberts, R., Skvortsova, V., Teal, P., Toni, D., Vandermaelen, C., Voigt, T., Weber, M., and Yoon, B. W. (2008) Aspirin and extended-release dipyridamole versus clopidogrel for recurrent stroke. N. Engl. J. Med. 359, 1238–1251
- 76Morabito, L., Montesinos, M. C., Schreibman, D. M., Balter, L., Thompson, L. F., Resta, R., Carlin, G., Huie, M. A., and Cronstein, B. N. (1998) Methotrexate and sulfasalazine promote adenosine release by a mechanism that requires ecto-5′-nucleotidase-mediated conversion of adenine nucleotides. J. Clin. Invest. 101, 295–300
- 77Cronstein, B. N. (1997) The mechanism of action of methotrexate. Rheum. Dis. Clin. North Am. 23, 739–755
- 78Cronstein, B. N., Naime, D., and Ostad, E. (1994) The antiinflammatory effects of methotrexate are mediated by adenosine. Adv. Exp. Med. Biol. 370, 411–416
- 79Cronstein, B. N., Naime, D., and Ostad, E. (1993) The antiinflammatory mechanism of methotrexate. Increased adenosine release at inflamed sites diminishes leukocyte accumulation in an in vivo model of inflammation. J. Clin. Invest. 92, 2675–2682
- 80Cronstein, B. N., Eberle, M. A., Gruber, H. E., and Levin, R. I. (1991) Methotrexate inhibits neutrophil function by stimulating adenosine release from connective tissue cells. Proc. Natl. Acad. Sci. U. S. A. 88, 2441–2445
- 81Eltzschig, H. K., Faigle, M., Knapp, S., Karhausen, J., Ibla, J., Rosenberger, P., Odegard, K. C., Laussen, P. C., Thompson, L. F., and Colgan, S. P. (2006) Endothelial catabolism of extracellular adenosine during hypoxia: the role of surface adenosine deaminase and CD26. Blood 108, 1602–1610
- 82Morote-Garcia, J. C., Rosenberger, P., Kuhlicke, J., and Eltzschig, H. K. (2008) HIF-1-dependent repression of adenosine kinase attenuates hypoxia-induced vascular leak. Blood 111, 5571–5580
- 83Sitkovsky, M. V. (2008) Damage control by hypoxia-inhibited AK. Blood 111, 5424–5425
- 84Jackson, E. K., and Gillespie, D. G. (2013) Extracellular 2′,3′-cAMP-adenosine pathway in proximal tubular, thick ascending limb, and collecting duct epithelial cells. Am. J. Physiol. Renal Physiol. 304, F49–F55
- 85Jackson, E. K., Ren, J., Gillespie, D. G., and Dubey, R. K. (2010) Extracellular 2′,3′-cyclic adenosine 5′-monophosphate is a potent inhibitor of preglomerular vascular smooth muscle cell and mesangial cell growth. Hypertension 56, 151–158
- 86Jackson, E. K., Ren, J., and Mi, Z. (2009) Extracellular 2′,3′-cAMP is a source of adenosine. J. Biol. Chem. 284, 33097–33106
- 87Yao, S. Y., Ng, A. M., Muzyka, W. R., Griffiths, M., Cass, C. E., Baldwin, S. A., and Young, J. D. (1997) Molecular cloning and functional characterization of nitrobenzylthioinosine (NBMPR)-sensitive (es) and NBMPR-insensitive (ei) equilibrative nucleoside transporter proteins (rENT1 and rENT2) from rat tissues. J. Biol. Chem. 272, 28423–28430