Redox gene therapy for ischemia/reperfusion injury of the liver reduces AP1 and NF-κB activation

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Liver transplantation is the only therapeutic strategy for many inherited and acquired diseases. The formation of reactive oxygen species following ischemia/reperfusion is a cause of hepatocellular injury during transplantation. This report describes the therapeutic application of mitochondrial superoxide dismutase gene transfer to the liver for acute ischemia/reperfusion injury. Recombinant adenoviral expression of mitochondrial superoxide dismutase in mouse liver prior to lobar ischemia/reperfusion significantly reduced acute liver damage and associated redox activation of both NF-κB and AP1. These immediate early transcription factors represent common pathways by which cells respond to environmental stress. This work provides the foundation for redox-mediated gene therapies directed at ameliorating ischemia/reperfusion injury and associated acute rejection in orthotopic liver transplantation.

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  1. 1

    Strassberg, S.M., Howard, T.K., Molmenti, E.P. & Hertl, M. Selecting the Donor Liver: Risk Factors for Poor Function After Orthotopic Liver Transplantation. Hepatology 20, 829–838 (1994).

  2. 2

    Kiuchi, T. et al. Backgrounds of early intragraft immune activation and rejection in liver transplant recipients. Impact of graft reperfusion quality. Transplantation 60, 49–55 (1995).

  3. 3

    Shaked, A., Nunes, F.A., Olthoff, K.M. & Lucey, M.R. Assessment of liver function: pre- and peritransplant evaluation. Clin. Chem. 43, 1539–1545 (1997).

  4. 4

    Goode, H.F. et al. Reperfusion injury,antioxidants and hemodynamics during orthotopic liver transplantation. Hepatology 19, 354–359 (1994).

  5. 5

    Arthur, M.J.P. et al. Oxygen-Derived Free Radicals Promote Hepatic Injury in the Rat. Gastroenterology 89, 114–1122 (1985).

  6. 6

    Jaeschke, H., Smith, C.V. & Mitchell, J.R. Hypoxic Damage Generates Reactive Oxygen Species in Isolated Perfused Rat Livers. Biochem. Biophys. Res. Commun. 150, 568–574 (1988).

  7. 7

    Colletti, L.M. et al. Role of Tumor Necrosis Factor-a in the Pathophysiologic Alterations after Hepatic Ischemia/Reperfusion Injury in the Rat. J. Clin. Invest. 85, 1936–1943 (1990).

  8. 8

    Colletti, L.M. et al. Chemokine Expression during Hepatic Ischemia/Reperfusion-indused Lung Injury in the Rat. J. Clin. Invest. 95, 134–141 (1995).

  9. 9

    Jaeschke, H., Farhood, A. & Smith, C.W. Neutrophils contribute to ischemia/reperfusion injury in rat liver in vivo . FASEB.J. 4, 3355–3359 (1990).

  10. 10

    Suzuki, S., Toledo-Pereyra, L.H. & Rodriquez, F.J. Role of neutrophils during the first 24 hours after liver ischemia and reperfusion Injury. Transplant. Proc. 26, 3695–3700 (1994).

  11. 11

    Atalla, S.L., Toledo-Pereyra, L.H., Mackenzie, G.H. & Cederna, J.P. Influence of oxygen-derived free radical scavengers on ischemia livers. Transplantation 40, 584–590 (1985).

  12. 12

    Mathews, W.R., Guido, D.M., Fisher, M.A. & Jaeschke, H. Lipid peroxidation as molecular mechanism of liver cell injury during reperfusion after ischemia. Free Radic. Biol. Med. 16, 763–770 (1994).

  13. 13

    Hunter, T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signaling. Cell 80, 225–236 (1995).

  14. 14

    Stamler, J.S. Redox signaling: nitrosylation and related target interactions of nitric oxide. Cell 78, 931–936 (1994).

  15. 15

    Bradham, C.A. et al. Reperfusion after liver transplantation in rats differentially activates the mitogen-activated protein kinases. Hepatology 25, 1128–1135 (1997).

  16. 16

    Schreck, R., Rieber, P. & Baeuerle, P.A. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. EMBO J. 10, 2247–2258 (1991).

  17. 17

    Karin, M., Liu, Z.G. & Zandi, E. AP-1 function and regulation. Curr.Op. Cell Biol. 9, 240–246 (1997).

  18. 18

    Baichwal, V.R. & Baeuerle, P.A. Activate NF-kappa B or die?. Curr. Biol. 7, R94–96 (1997)

  19. 19

    Colotta, F., Polentarutti, N., Sironi, M. & Mantovani, A. Expression and involvement of c-fos and c-jun protooncogenes in programmed cell death induced by growth factor deprivation in lymphoid cell lines. J. Biol. Chem. 267, 18278–18283 (1992).

  20. 20

    Roffler-Tarlov, S. et al. Programmed cell death in the absence of c-Fos and c-Jun. Development 122, 1–9 (1996).

  21. 21

    Westwick, J.K., Weitzel, C., Leffert, H.L. & Brenner, D.A. Activation of Jun kinase is an early event in hepatic regeneration. J. Clin. Invest. 95, 803–810 (1995).

  22. 22

    Schlossberg, H., Zhang, Y., Dudus, L., & Engelhardt, J.F. Expression of c-fos and c-jun During Hepatocellular Remodeling Following Ischemia Reperfusion in Mouse Liver Hepatology 23, 1546–1555 (1996).

  23. 23

    Pfarr, C.M. et al. Mouse JunD negatively regulates fibroblast growth and antagonizes transformation by ras. Cell 76, 747–760 (1994).

  24. 24

    Engelhardt, J.F., Ye, X., Doranz, B. & Wilson, J.M. Second Generation Recombinant Adenoviral Vectors Improves Transgene Persistence and Decreases Immune Response in Mouse Liver Proc. Natl. Acad. Sci. USA 91, 6196–6200 (1994).

  25. 25

    Zwacka, R.M. et al. T-Lymphocyte Mediated Ischemia/Reperfusion Induced Inflammatory responses in Mouse Liver J.Clin. Inv. 100, 279–289 (1997).

  26. 26

    Yang, Y., Ertl, H.C. & Wilson, J.M. MHC class I-restricted cytotoxic T lymphocytes to viral antigens destroy hepatocytes in mice infected with E1-deleted recombinant adenoviruses. Immunity 1, 433–442 (1994).

  27. 27

    Baeuerle, P.A. The inducible transcription activator NF-κB: regulation by distinct protein subunits. Biochem. Biophys. Act 1072, 63–80 (1991).

  28. 28

    Abate, C., Patel, L., Rauscher, F.J. & Curran, T. Redox regulation of fos and jun DNA-binding activity in vitro. Science 249, 1157–1161 (1990).

  29. 29

    Imbert, V. et al. Tyrosine phosphorylation of I kappa B-alpha activates NF-kappa B without proteolytic degradation of I kappa B-alpha. Cell 86, 787–798 (1996).

  30. 30

    Alkalay, I. et al. Stimulation-dependent IκBα phosphorylation marks the NF-κB inhibitor for degradation via the ubiquitin-proteasome pathway. Proc. Natl. Acad. Sci. USA 92, 10599–10603 (1995).

  31. 31

    Schreck, R., Meier, B., Mannel, D.N., Droge, W., & Baeuerle, P.A. Dithiocarbamates as potent inhibitors of nuclear factor kappa B activation J. Exp. Med. 175, 1181–94 (1992).

  32. 32

    Schulze-Osthoff, K. et al. Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation. J. Biol. Chem. 267, 5317–5323 (1992).

  33. 33

    Schulze-Osthoff, K., Beyaert, R., Vandevborde, V., Haegeman, G., & Fiers, W. Depletion of the mitochondrial electron transport abrogates the cytotoxic and gene-inductive effects of TNF. EMBO J. 12, 3095–3104 (1993).

  34. 34

    Okuaki, Y. et al. Splenectomy-reduced hepatic injury induced by ischemia/reperfusion in the rat. Liver 16, 188–196 (1996).

  35. 35

    Lee, F.S., Hagler, J., Chen, Z.J., & Maniatis, T. Activation of the IkappaB alpha kinase complex by MEKK1, a kinase of the JNK pathway. Cell 88, 213–222 (1997).

  36. 36

    Zwacka, R.M., Dudus, L., Epperly, M.W., Greenberger, J.S. & Engelhardt, J.F. Redox gene therapy protects against Ionizing radiation induced apoptosis. Human Gene Therapy (in the press).

  37. 37

    Engelhardt, J.F. Methods of Adenoviral-Mediated Gene Transfer to Airway Epithelium in Methods in Molecular Biology: Gene Therapy Protocols 169–184 (Humana Press, Clifton, NJ, 1996).

  38. 38

    Oberley, T.D., Oberley, L.W., Slattery, A.F., Lauchner, L.J. & Elwell, J.H. Immunohistochemical localization of antioxidant enzymes in adult Syrian hamster tissues and during kidney development. Am. J. Pathol. 137, 199–214 (1990).

  39. 39

    Beauchamp, C. & Fridovich, I. Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44, 276–287 (1971)

  40. 40

    Zhong, W. et al. Inhibition of cell growth and sensitization to oxidative damage by overexpression of manganese superoxide dismutase in rat glioma cells. Cell Growth Differ. 7, 1175–1186 (1996).

  41. 41

    Hattori, M., Tugores, A., Karin, M. & Brenner, D.A. A simplified method for the preparation of nuclear extracts. DNA 9, 777 (1990).

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Zwacka, R., Zhou, W., Zhang, Y. et al. Redox gene therapy for ischemia/reperfusion injury of the liver reduces AP1 and NF-κB activation. Nat Med 4, 698–704 (1998) doi:10.1038/nm0698-698

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