Intracellular NAD levels regulate tumor necrosis factor protein synthesis in a sirtuin-dependent manner (original) (raw)

Nature Medicine volume 15, pages 206–210 (2009)Cite this article

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.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 12 print issues and online access

$209.00 per year

only $17.42 per issue

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

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).
    Article CAS Google Scholar
  2. Henson, P.M. Dampening inflammation. Nat. Immunol. 6, 1179–1181 (2005).
    Article CAS 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).
    Article CAS Google Scholar
  4. Saunders, L.R. & Verdin, E. Sirtuins: critical regulators at the crossroads between cancer and aging. Oncogene 26, 5489–5504 (2007).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS Google Scholar
  22. Mostoslavsky, R. et al. Genomic instability and aging-like phenotype in the absence of mammalian SIRT6. Cell 124, 315–329 (2006).
    Article CAS Google Scholar
  23. Michishita, E. et al. SIRT6 is a histone H3 lysine 9 deacetylase that modulates telomeric chromatin. Nature 452, 492–496 (2008).
    Article CAS 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).
    Article CAS 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).
    Article CAS 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).
    Article CAS Google Scholar

Download references

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.

Author information

Author notes

  1. Pierre-Paul Prevot, Raul Mostoslavsky & Thibaut De Smedt
    Present address: Present addresses: Université Catholique de Louvain, Christian de Duve Institute of Cellular Pathology Hormone and Metabolic Research Unit, Avenue Hippocrate 75 /7529, B-1200 Brussels, Belgium (P.-P.P.); Department of Medicine, Massachusetts General Hospital Cancer Center, Harvard Medical School, 185 Cambridge Street, Boston, Massachusetts 02114, USA (R.M.); Addex Pharmaceuticals, 12 chemin des Aulx Plan-Les-Ouates, CH-1228 Geneva, Switzerland (T.D.S.).,
  2. Frédéric Van Gool and Mara Gallí: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratoire de Physiologie Animale, Institut de Biologie et Médecine Moléculaire, Université Libre de Bruxelles, 12 rue des Professeur Jeneer et Brachet, Gosselies, 6041, Belgium
    Frédéric Van Gool, Mara Gallí & Oberdan Leo
  2. Laboratoire de Chimie Biologique, Institut de Biologie et Médecine Moléculaire, Université Libre de Bruxelles, 12 rue des Professeur Jeneer et Brachet, Gosselies, 6041, Belgium
    Cyril Gueydan & Véronique Kruys
  3. Laboratory for Molecular Biology of Ectoparasites, Institut de Biologie et Médecine Moléculaire, Université Libre de Bruxelles, 12 rue des Professeur Jeneer et Brachet, Gosselies, 6041, Belgium
    Pierre-Paul Prevot
  4. Clinical Research Division, Fred Hutchinson Cancer Research Center, D2-100, 1100 Fairview Avenue North, Seattle, 98109, Washington, USA
    Antonio Bedalov
  5. The Center for Blood Research and Department of Genetics, Howard Hughes Medical Institute, The Children's Hospital, Harvard University Medical School, 300 Longwood Avenue, Boston, 02115, Massachusetts, USA
    Raul Mostoslavsky & Frederick W Alt
  6. TopoTarget Switzerland S.A., Avenue de Sévelin 18-20, Lausanne, CH-1004, Switzerland
    Thibaut De Smedt

Authors

  1. Frédéric Van Gool
    You can also search for this author inPubMed Google Scholar
  2. Mara Gallí
    You can also search for this author inPubMed Google Scholar
  3. Cyril Gueydan
    You can also search for this author inPubMed Google Scholar
  4. Véronique Kruys
    You can also search for this author inPubMed Google Scholar
  5. Pierre-Paul Prevot
    You can also search for this author inPubMed Google Scholar
  6. Antonio Bedalov
    You can also search for this author inPubMed Google Scholar
  7. Raul Mostoslavsky
    You can also search for this author inPubMed Google Scholar
  8. Frederick W Alt
    You can also search for this author inPubMed Google Scholar
  9. Thibaut De Smedt
    You can also search for this author inPubMed Google Scholar
  10. Oberdan Leo
    You can also search for this author inPubMed Google Scholar

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.

Corresponding author

Correspondence toOberdan Leo.

Ethics declarations

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.

Supplementary information

Rights and permissions

About this article

Cite this article

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

Download citation

This article is cited by