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Intracellular NAD levels regulate tumor necrosis factor protein synthesis in a sirtuin-dependent manner

Abstract

Tumor necrosis factor (TNF) synthesis is known to play a major part in numerous inflammatory disorders, and multiple transcriptional and post-transcriptional regulatory mechanisms have therefore evolved to dampen the production of this key proinflammatory cytokine1,2. The high expression of nicotinamide phosphoribosyltransferase (Nampt), an enzyme involved in the nicotinamide-dependent NAD biosynthetic pathway, in cells of the immune system3 has led us to examine the potential relationship between NAD metabolism and inflammation. We show here that intracellular NAD concentration promotes TNF synthesis by activated immune cells. Using a positive screen, we have identified Sirt6, a member of the sirtuin family4, as the NAD-dependent enzyme able to regulate TNF production by acting at a post-transcriptional step. These studies reveal a previously undescribed relationship between metabolism and the inflammatory response and identify Sirt6 and the nicotinamide-dependent NAD biosynthetic pathway as novel candidates for immunointervention in an inflammatory setting.

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Figure 1: Intracellular NAD levels determine TNF production capacities.
Figure 2: Immunomodulatory properties of sirtuin inhibitors.
Figure 3: Sirtuin inhibitors affect TNF production at a post-transcriptional step.
Figure 4: SIRT6 regulates TNF protein synthesis.

References

  1. Feldmann, M. & Maini, R.N. Lasker Clinical Medical Research Award. TNF defined as a therapeutic target for rheumatoid arthritis and other autoimmune diseases. Nat. Med. 9, 1245–1250 (2003).

    CAS  Article  Google Scholar 

  2. Henson, P.M. Dampening inflammation. Nat. Immunol. 6, 1179–1181 (2005).

    CAS  Article  Google Scholar 

  3. Luk, T., Malam, Z. & Marshall, J.C. Pre-B cell colony-enhancing factor (PBEF)/visfatin: a novel mediator of innate immunity. J. Leukoc. Biol. 83, 804–816 (2008).

    CAS  Article  Google Scholar 

  4. Saunders, L.R. & Verdin, E. Sirtuins: critical regulators at the crossroads between cancer and aging. Oncogene 26, 5489–5504 (2007).

    CAS  Article  Google Scholar 

  5. Ziegler, M. New functions of a long-known molecule. Emerging roles of NAD in cellular signaling. Eur. J. Biochem. 267, 1550–1564 (2000).

    CAS  Article  Google Scholar 

  6. Berger, F., Ramirez-Hernandez, M.H. & Ziegler, M. The new life of a centenarian: signalling functions of NAD(P). Trends Biochem. Sci. 29, 111–118 (2004).

    CAS  Article  Google Scholar 

  7. Schreiber, V., Dantzer, F., Ame, J.C. & de Murcia, G. Poly(ADP-ribose): novel functions for an old molecule. Nat. Rev. Mol. Cell Biol. 7, 517–528 (2006).

    CAS  Article  Google Scholar 

  8. Rongvaux, A. et al. Pre-B-cell colony-enhancing factor, whose expression is up-regulated in activated lymphocytes, is a nicotinamide phosphoribosyltransferase, a cytosolic enzyme involved in NAD biosynthesis. Eur. J. Immunol. 32, 3225–3234 (2002).

    CAS  Article  Google Scholar 

  9. Jia, S.H. et al. Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. J. Clin. Invest. 113, 1318–1327 (2004).

    CAS  Article  Google Scholar 

  10. Ye, S.Q. et al. Pre-B-cell colony-enhancing factor as a potential novel biomarker in acute lung injury. Am. J. Respir. Crit. Care Med. 171, 361–370 (2005).

    Article  Google Scholar 

  11. Busso, N. et al. Pharmacological inhibition of nicotinamide phosphoribosyltransferase/visfatin enzymatic activity identifies a new inflammatory pathway linked to NAD. PLoS ONE 3, e2267 (2008).

    Article  Google Scholar 

  12. Fukuzawa, M. et al. Inhibitory effect of nicotinamide on in vitro and in vivo production of tumor necrosis factor-α. Immunol. Lett. 59, 7–11 (1997).

    CAS  Article  Google Scholar 

  13. Ungerstedt, J.S., Blomback, M. & Soderstrom, T. Nicotinamide is a potent inhibitor of proinflammatory cytokines. Clin. Exp. Immunol. 131, 48–52 (2003).

    CAS  Article  Google Scholar 

  14. Szabo, C. Nicotinamide: a jack of all trades (but master of none?). Intensive Care Med. 29, 863–866 (2003).

    Article  Google Scholar 

  15. Cuzzocrea, S. Shock, inflammation and PARP. Pharmacol. Res. 52, 72–82 (2005).

    CAS  Article  Google Scholar 

  16. Hassa, P.O. & Hottiger, M.O. The functional role of poly(ADP-ribose)polymerase 1 as novel coactivator of NF-κB in inflammatory disorders. Cell. Mol. Life Sci. 59, 1534–1553 (2002).

    CAS  Article  Google Scholar 

  17. Oliver, F.J. et al. Resistance to endotoxic shock as a consequence of defective NF-κB activation in poly (ADP-ribose) polymerase-1 deficient mice. EMBO J. 18, 4446–4454 (1999).

    CAS  Article  Google Scholar 

  18. Kuhnle, S., Nicotera, P., Wendel, A. & Leist, M. Prevention of endotoxin-induced lethality, but not of liver apoptosis in poly(ADP-ribose) polymerase–deficient mice. Biochem. Biophys. Res. Commun. 263, 433–438 (1999).

    CAS  Article  Google Scholar 

  19. Hasmann, M. & Schemainda, I. FK866, a highly specific noncompetitive inhibitor of nicotinamide phosphoribosyltransferase, represents a novel mechanism for induction of tumor cell apoptosis. Cancer Res. 63, 7436–7442 (2003).

    CAS  PubMed  Google Scholar 

  20. Grozinger, C.M., Chao, E.D., Blackwell, H.E., Moazed, D. & Schreiber, S.L. Identification of a class of small molecule inhibitors of the sirtuin family of NAD-dependent deacetylases by phenotypic screening. J. Biol. Chem. 276, 38837–38843 (2001).

    CAS  Article  Google Scholar 

  21. Heltweg, B. et al. Antitumor activity of a small-molecule inhibitor of human silent information regulator 2 enzymes. Cancer Res. 66, 4368–4377 (2006).

    CAS  Article  Google Scholar 

  22. Mostoslavsky, R. et al. Genomic instability and aging-like phenotype in the absence of mammalian SIRT6. Cell 124, 315–329 (2006).

    CAS  Article  Google Scholar 

  23. Michishita, E. et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452, 492–496 (2008).

    CAS  Article  Google Scholar 

  24. Ulloa, L. et al. Ethyl pyruvate prevents lethality in mice with established lethal sepsis and systemic inflammation. Proc. Natl. Acad. Sci. USA 99, 12351–12356 (2002).

    CAS  Article  Google Scholar 

  25. Inaba, K. et al. Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor. J. Exp. Med. 176, 1693–1702 (1992).

    CAS  Article  Google Scholar 

  26. Zerez, C.R., Lee, S.J. & Tanaka, K.R. Spectrophotometric determination of oxidized and reduced pyridine nucleotides in erythrocytes using a single extraction procedure. Anal. Biochem. 164, 367–373 (1987).

    CAS  Article  Google Scholar 

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Acknowledgements

APO866 was synthesized and kindly provided by Astellas Pharma. We wish to thank G. de Murcia (formerly at the Ecole Supérieure de Biotechnologie de Strasbourg, Illkirch Cedex) for providing the Parp1-knockout mouse strain and V. Sartorelli (US National Institutes of Health) for providing the pHan-SIRT1 vector (wild-type and mutant forms). This work was supported by The Belgian Program in Interuniversity Poles of Attraction initiated by the Belgian state, the Prime Minister's office, Science Policy Programming, by a Research Concerted Action of the Communauté française de Belgique, by grants from the Direction Générale des Technologies de la Recherche et de l'Energie, Région Wallonne (Belgium), by a grant from the Fonds Jean Brachet and by TopoTarget Switzerland SA. F.V.G. and M.G. have been supported by research grants from the Fonds national de la recherché scientifique (FRS-FNRS), French community of Belgium. The scientific responsibility is assumed by the authors.

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Authors

Contributions

F.V.G. and M.G. generated all of the in vitro data presented in the manuscript. V.K. and C.G. designed and provided assistance for the polysome purification and the northern blot experiments and helped with the analysis of the data. P.-P.P. and F.V.G. designed and performed the in vivo experiments. A.B. and T.D.S. provided the pharmacological reagents used in this study (cambinol and APO866, respectively). R.M. and F.W.A. provided bone marrow cells from Sirt6-knockout mice. F.V.G. and O.L. designed the study and analyzed the data. O.L. directed the project and wrote the manuscript.

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Correspondence to Oberdan Leo.

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Competing interests

T.D.S. was a paid employee of TopoTarget Switzerland S.A. during the completion of this work. Part of the work was performed under a research grant provided by TopoTarget. F.V.G., M.G., O.L. and T.D.S. have made patent applications to the World Intellectual Property Organization (WIPO) pertaining to the possible use of APO866 to treat inflammatory-related disorders.

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Van Gool, F., Gallí, M., Gueydan, C. et al. Intracellular NAD levels regulate tumor necrosis factor protein synthesis in a sirtuin-dependent manner. Nat Med 15, 206–210 (2009). https://doi.org/10.1038/nm.1906

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