Abstract
Here we report that bufalin and other cardiac glycoside inhibitors of the sodium-potassium ATPase (sodium pump) potently inhibit the induction of the interferon-β (IFNβ) gene by virus, double-stranded RNA or double-stranded DNA. Cardiac glycosides increase the intracellular sodium concentration, which appears to inhibit the ATPase activity of the RNA sensor RIG-I, an essential and early component in the IFNβ activation pathway. This, in turn, prevents the activation of the critical transcription factors IRF3 and NFκB. Bufalin inhibition can be overcome by expressing a drug-resistant variant of the sodium pump and knocking down the pump by short hairpin RNA inhibits IFNβ expression. Thus, bufalin acts exclusively through the sodium pump. We also show that bufalin inhibits tumor necrosis factor (TNF) signaling, at least in part by interfering with the nuclear translocation of NFκB. These findings suggest that bufalin could be used to treat inflammatory and autoimmune diseases in which IFN or TNF are hyperactivated.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Sen, G.C. Viruses and interferons. Annu. Rev. Microbiol. 55, 255–281 (2001).
Honda, K., Takaoka, A. & Taniguchi, T. Type I interferon [corrected] gene induction by the interferon regulatory factor family of transcription factors. Immunity 25, 349–360 (2006).
Chiu, Y.H., Macmillan, J.B. & Chen, Z.J. RNA polymerase III detects cytosolic DNA and induces type I interferons through the RIG-I pathway. Cell 138, 576–591 (2009).
Ablasser, A. et al. RIG-I-dependent sensing of poly(dA:dT) through the induction of an RNA polymerase III-transcribed RNA intermediate. Nat. Immunol. 10, 1065–1072 (2009).
García-Sastre, A. & Biron, C.A. Type 1 interferons and the virus-host relationship: a lesson in detente. Science 312, 879–882 (2006).
Le Bon, A. & Tough, D.F. Links between innate and adaptive immunity via type I interferon. Curr. Opin. Immunol. 14, 432–436 (2002).
Banchereau, J. & Pascual, V. Type I interferon in systemic lupus erythematosus and other autoimmune diseases. Immunity 25, 383–392 (2006).
Hall, J.C. & Rosen, A. Type I interferons: crucial participants in disease amplification in autoimmunity. Nat. Rev. Rheumatol. 6, 40–49 (2010).
Mandl, J.N. et al. Divergent TLR7 and TLR9 signaling and type I interferon production distinguish pathogenic and nonpathogenic AIDS virus infections. Nat. Med. 14, 1077–1087 (2008).
Whittemore, L.A. & Maniatis, T. Postinduction turnoff of beta-interferon gene expression. Mol. Cell. Biol. 10, 1329–1337 (1990).
Jacquelin, B. et al. Nonpathogenic SIV infection of African green monkeys induces a strong but rapidly controlled type I IFN response. J. Clin. Invest. 119, 3544–3555 (2009).
Maniatis, T. et al. Structure and function of the interferon-beta enhanceosome. Cold Spring Harb. Symp. Quant. Biol. 63, 609–620 (1998).
Sun, L., Liu, S. & Chen, Z.J. SnapShot: pathways of antiviral innate immunity. Cell 140, 436–436.e2 (2010).
Ford, E. & Thanos, D. The transcriptional code of human IFN-beta gene expression. Biochim. Biophys. Acta 1799, 328–336 (2010).
Kato, H. et al. Differential roles of MDA5 and RIG-I helicases in the recognition of RNA viruses. Nature 441, 101–105 (2006).
Cui, S. et al. The C-terminal regulatory domain is the RNA 5′-triphosphate sensor of RIG-I. Mol. Cell 29, 169–179 (2008).
Takahasi, K. et al. Nonself RNA-sensing mechanism of RIG-I helicase and activation of antiviral immune responses. Mol. Cell 29, 428–440 (2008).
Seth, R.B., Sun, L., Ea, C.K. & Chen, Z.J. Identification and characterization of MAVS, a mitochondrial antiviral signaling protein that activates NF-kappaB and IRF 3. Cell 122, 669–682 (2005).
Tang, E.D. & Wang, C.Y. MAVS self-association mediates antiviral innate immune signaling. J. Virol. 83, 3420–3428 (2009).
Fujita, T. A nonself RNA pattern: tri-p to panhandle. Immunity 31, 4–5 (2009).
Myong, S. et al. Cytosolic viral sensor RIG-I is a 5′-triphosphate-dependent translocase on double-stranded RNA. Science 323, 1070–1074 (2009).
Gack, M.U. et al. TRIM25 RING-finger E3 ubiquitin ligase is essential for RIG-I-mediated antiviral activity. Nature 446, 916–920 (2007).
Gack, M.U., Nistal-Villan, E., Inn, K.S., Garcia-Sastre, A. & Jung, J.U. Phosphorylation-mediated negative regulation of RIG-I antiviral activity. J. Virol. 84, 3220–3229 (2010).
Saha, S.K. et al. Regulation of antiviral responses by a direct and specific interaction between TRAF3 and Cardif. EMBO J. 25, 3257–3263 (2006).
Guo, B. & Cheng, G. Modulation of the interferon antiviral response by the TBK1/IKKi adaptor protein TANK. J. Biol. Chem. 282, 11817–11826 (2007).
Kawai, T. et al. IPS-1, an adaptor triggering RIG-I- and Mda5-mediated type I interferon induction. Nat. Immunol. 6, 981–988 (2005).
Ishikawa, H. & Barber, G.N. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674–678 (2008).
Zhong, B. et al. The adaptor protein MITA links virus-sensing receptors to IRF3 transcription factor activation. Immunity 29, 538–550 (2008).
Schröder, M., Baran, M. & Bowie, A.G. Viral targeting of DEAD box protein 3 reveals its role in TBK1/IKKepsilon-mediated IRF activation. EMBO J. 27, 2147–2157 (2008).
Prassas, I. & Diamandis, E.P. Novel therapeutic applications of cardiac glycosides. Nat. Rev. Drug Discov. 7, 926–935 (2008).
Hemmi, H. et al. The roles of two IkappaB kinase-related kinases in lipopolysaccharide and double stranded RNA signaling and viral infection. J. Exp. Med. 199, 1641–1650 (2004).
Fitzgerald, K.A. et al. IKKepsilon and TBK1 are essential components of the IRF3 signaling pathway. Nat. Immunol. 4, 491–496 (2003).
Yoneyama, M. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat. Immunol. 5, 730–737 (2004).
Saito, T. et al. Regulation of innate antiviral defenses through a shared repressor domain in RIG-I and LGP2. Proc. Natl. Acad. Sci. USA 104, 582–587 (2007).
Langer, G.A. Ionic basis of myocardial contractility. Annu. Rev. Med. 28, 13–20 (1977).
Lingrel, J.B. The physiological significance of the cardiotonic steroid/ouabain-binding site of the Na,K-ATPase. Annu. Rev. Physiol. 72, 395–412 (2010).
Ohtsubo, M., Noguchi, S., Takeda, K., Morohashi, M. & Kawamura, M. Site-directed mutagenesis of Asp-376, the catalytic phosphorylation site, and Lys-507, the putative ATP-binding site, of the alpha-subunit of Torpedo californica Na+/K(+)-ATPase. Biochim. Biophys. Acta 1021, 157–160 (1990).
Simpson, C.D. et al. Inhibition of the sodium potassium adenosine triphosphatase pump sensitizes cancer cells to anoikis and prevents distant tumor formation. Cancer Res. 69, 2739–2747 (2009).
Izquierdo, I. Nimodipine and the recovery of memory. Trends Pharmacol. Sci. 11, 309–310 (1990).
Trube, G., Rorsman, P. & Ohno-Shosaku, T. Opposite effects of tolbutamide and diazoxide on the ATP-dependent K+ channel in mouse pancreatic beta-cells. Pflugers Arch. 407, 493–499 (1986).
Garvin, J.L., Simon, S.A., Cragoe, E.J. Jr. & Mandel, L.J. Phenamil: an irreversible inhibitor of sodium channels in the toad urinary bladder. J. Membr. Biol. 87, 45–54 (1985).
Xie, Z. & Cai, T. Na+-K+–ATPase-mediated signal transduction: from protein interaction to cellular function. Mol. Interv. 3, 157–168 (2003).
Gee, P. et al. Essential role of the N-terminal domain in the regulation of RIG-I ATPase activity. J. Biol. Chem. 283, 9488–9496 (2008).
Cyert, M.S. Regulation of nuclear localization during signaling. J. Biol. Chem. 276, 20805–20808 (2001).
Yang, Q. et al. Cardiac glycosides inhibit TNF-alpha/NF-kappaB signaling by blocking recruitment of TNF receptor-associated death domain to the TNF receptor. Proc. Natl. Acad. Sci. USA 102, 9631–9636 (2005).
Ronnblom, L. & Elkon, K.B. Cytokines as therapeutic targets in SLE. Nat. Rev. Rheumatol. 6, 339–347 (2010).
Meng, Z. et al. Pilot study of huachansu in patients with hepatocellular carcinoma, nonsmall-cell lung cancer, or pancreatic cancer. Cancer 115, 5309–5318 (2009).
Chen, S. et al. A small molecule that directs differentiation of human ESCs into the pancreatic lineage. Nat. Chem. Biol. 5, 258–265 (2009).
Acknowledgements
This work is supported by US National Institutes of Health grant 5R01AI020642-26 (to T.M.). S.C. is supported by the postdoctoral fellowship from Juvenile Diabetes Research Foundation. We thank X. Sun (Brandeis University) for help with the microarray analysis. We also thank S.-L. Ng for critical reading of this manuscript and S. Schalm for helpful discussions.
Author information
Authors and Affiliations
Contributions
J.Y., S.C. and T.M. conceived the research, J.Y. and S.C. conducted experiments, and J.Y. and T.M. wrote the paper.
Corresponding author
Ethics declarations
Competing interests
The authors have filed a patent on the inhibition of interferon-β expression by cardiac glycosides described in the paper.
Supplementary information
Supplementary Text and Figures
Supplementary Methods, Supplementary Table 1 and Supplementary Figures 1–12 (PDF 13254 kb)
Rights and permissions
About this article
Cite this article
Ye, J., Chen, S. & Maniatis, T. Cardiac glycosides are potent inhibitors of interferon-β gene expression. Nat Chem Biol 7, 25–33 (2011). https://doi.org/10.1038/nchembio.476
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nchembio.476
This article is cited by
-
ATP1A1/BCL2L1 predicts the response of myelomonocytic and monocytic acute myeloid leukemia to cardiac glycosides
Leukemia (2024)
-
Near-infrared light-controllable bufalin delivery from a black phosphorus-hybrid supramolecular hydrogel for synergistic photothermal-chemo tumor therapy
Nano Research (2021)
-
Convallatoxin enhance the ligand-induced mu-opioid receptor endocytosis and attenuate morphine antinociceptive tolerance in mice
Scientific Reports (2019)
-
Toll-like receptor 3 activation selectively reverses HIV latency in microglial cells
Retrovirology (2017)
-
Signaling function of Na,K-ATPase induced by ouabain against LPS as an inflammation model in hippocampus
Journal of Neuroinflammation (2014)