[en] IL-1beta and TNF-alpha contribute to pancreatic beta cell death in type 1 diabetes. Both cytokines activate the transcription factor nuclear factor-kappaB (NF-kappaB), but recent observations suggest that NF-kappaB blockade prevents IL-1beta + IFN-gamma- but not TNF-alpha + IFN-gamma-induced beta cell apoptosis. The aim of the present study was to compare the effects of IL-1beta and TNF-alpha on cell death and the pattern of NF-kappaB activation and global gene expression in beta cells. METHODS: Cell viability was measured after exposure to IL-1beta or to TNF-alpha alone or in combination with IFN-gamma, and blockade of NF-kappaB activation or protein synthesis. INS-1E cells exposed to IL-1beta or TNF-alpha in time course experiments were used for IkappaB kinase (IKK) activation assay, detection of p65 NF-kappaB by immunocytochemistry, real-time RT-PCR and microarray analysis. RESULTS: Blocking NF-kappaB activation protected beta cells against IL-1beta + IFNgamma- or TNFalpha + IFNgamma-induced apoptosis. Blocking de novo protein synthesis did not increase TNF-alpha- or IL-1beta-induced beta cell death, in line with the observations that cytokines induced the expression of the anti-apoptotic genes A20, Iap-2 and Xiap to a similar extent. Microarray analysis of INS-1E cells treated with IL-1beta or TNF-alpha showed similar patterns of gene expression. IL-1beta, however, induced a higher rate of expression of NF-kappaB target genes putatively involved in beta cell dysfunction and death and a stronger activation of the IKK complex, leading to an earlier translocation of NF-kappaB to the nucleus. CONCLUSIONS/INTERPRETATION: NF-kappaB activation in beta cells has a pro-apoptotic role following exposure not only to IL-1beta but also to TNF-alpha. The more marked beta cell death induced by IL-1beta is explained at least in part by higher intensity NF-kappaB activation, leading to increased transcription of key target genes.
Research Center/Unit :
Giga-Signal Transduction - ULiège
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Ortis, Fernanda; Université Libre de Bruxelles - ULB > Laboratory of Experimental Medicine
Pirot, P.; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Génétique générale et humaine
Naamane, N.; Université de Liège - ULiège > Département de pharmacie > Chimie médicale
Thomas HE, Kay TW (2000) Beta cell destruction in the development of autoimmune diabetes in the non-obese diabetic (NOD) mouse. Diabetes Metab Res Rev 16:251-261
Eizirik DL, Mandrup-Poulsen T (2001) A choice of death-the signal-transduction of immune-mediated beta-cell apoptosis. Diabetologia 44:2115-2133
Cnop M, Welsh N, Jonas JC et al (2005) Mechanisms of pancreatic beta cell death in type 1 and type 2 diabetes: many differences, few similarities. Diabetes 54(Suppl 2):S97-S107
Ortis F, Cardozo AK, Crispim D, Storling J, Mandrup-Poulsen T, Eizirik DL (2006) Cytokine-induced pro-apoptotic gene expression in insulin-producing cells is related to rapid, sustained and non-oscillatory NF-κB activation. Mol Endocrinol 20:1867-1879
Giannoukakis N, Rudert WA, Trucco M, Robbins PD (2000) Protection of human islets from the effects of interleukin-1β by adenoviral gene transfer of an IκB repressor. J Biol Chem 275:36509-36513
Heimberg H, Heremans Y, Jobin C et al (2001) Inhibition of cytokine-induced NF-κB activation by adenovirus-mediated expression of a NF-κB super-repressor prevents beta cell apoptosis. Diabetes 50:2219-2224
Eldor R, Yeffet A, Baum K et al (2006) Conditional and specific NF-κB blockade protects pancreatic beta cells from diabetogenic agents. Proc Natl Acad Sci U S A 103:5072-5077
Cardozo AK, Heimberg H, Heremans Y et al (2001) A comprehensive analysis of cytokine-induced and nuclear factor-κB-dependent genes in primary rat pancreatic beta-cells. J Biol Chem 276:48879-48886
Cardozo AK, Ortis F, Storling J et al (2005) Cytokines downregulate the sarcoendoplasmic reticulum pump Ca2+ ATPase 2b and deplete endoplasmic reticulum Ca2+, leading to induction of endoplasmic reticulum stress in pancreatic beta cells. Diabetes 54:452-461
Pirot P, Ortis F, Cnop M et al (2007) Transcriptional regulation of the endoplasmic reticulum stress gene Chop in pancreatic insulin-producing cells. Diabetes 56:1069-1077
Eizirik DL, Cardozo AK, Cnop M (2008) The role for endoplasmic reticulum stress in diabetes mellitus. Endocr Rev 29:42-61
Karin M, Lin A (2002) NF-κB at the crossroads of life and death. Nat Immunol 3:221-227
Dutta J, Fan Y, Gupta N, Fan G, Gelinas C (2006) Current insights into the regulation of programmed cell death by NF-κB. Oncogene 25:6800-6816
Kim S, Millet I, Kim HS et al (2007) NF-κB prevents beta cell death and autoimmune diabetes in NOD mice. Proc Natl Acad Sci U S A 104:1913-1918
Hoffmann A, Leung TH, Baltimore D (2003) Genetic analysis of NF-κB/Rel transcription factors defines functional specificities. EMBO J 22:5530-5539
Saccani S, Pantano S, Natoli G (2003) Modulation of NF-κB activity by exchange of dimers. Mol Cell 11:1563-1574
Hayden MS, Ghosh S (2004) Signaling to NF-κB. Genes Dev 18:2195-2224
Hayden MS, Ghosh S (2008) Shared principles in NF-κB signaling. Cell 132:344-362
Vermeulen L, De Wilde G, Notebaert S, Van den Berghe W, Haegeman G (2002) Regulation of the transcriptional activity of the nuclear factor-κB p65 subunit. Biochem Pharmacol 64:963-970
Andersen NA, Larsen CM, Mandrup-Poulsen T (2000) TNFα and IFNγ potentiate IL-1β induced mitogen activated protein kinase activity in rat pancreatic islets of Langerhans. Diabetologia 43:1389-1396
Larsen L, Storling J, Darville M et al (2005) Extracellular signal-regulated kinase is essential for interleukin-1-induced and nuclear factor κB-mediated gene expression in insulin-producing INS-1E cells. Diabetologia 48:2582-2590
Beg AA, Baltimore D (1996) An essential role for NF-κB in preventing TNF-α-induced cell death. Science 274:782-784
Van Antwerp DJ, Martin SJ, Kafri T, Green DR, Verma IM (1996) Suppression of TNF-α-induced apoptosis by NF-κB. Science 274:787-789
Muppidi JR, Tschopp J, Siegel RM (2004) Life and death decisions: secondary complexes and lipid rafts in TNF receptor family signal transduction. Immunity 21:461-465
Pipeleers DG, in't Veld PA, Van de Winkel M, Maes E, Schuit FC, Gepts W (1985) A new in vitro model for the study of pancreatic A and B cells. Endocrinology 117:806-816
Rasschaert J, Ladriere L, Urbain M et al (2005) Toll-like receptor 3 and STAT-1 contribute to double-stranded RNA + interferon-γ-induced apoptosis in primary pancreatic beta-cells. J Biol Chem 280:33984-33991
Janjic D, Maechler P, Sekine N, Bartley C, Annen AS, Wolheim CB (1999) Free radical modulation of insulin release in INS-1 cells exposed to alloxan. Biochem Pharmacol 57:639-648
Kutlu B, Cardozo AK, Darville MI et al (2003) Discovery of gene networks regulating cytokine-induced dysfunction and apoptosis in insulin-producing INS-1 cells. Diabetes 52:2701-2719
Cardozo AK, Kruhoffer M, Leeman R, Orntoft T, Eizirik DL (2001) Identification of novel cytokine-induced genes in pancreatic beta-cells by high-density oligonucleotide arrays. Diabetes 50:909-920
Cnop M, Ladriere L, Hekerman P et al (2007) Selective inhibition of eukaryotic translation initiation factor 2 alpha dephosphorylation potentiates fatty acid-induced endoplasmic reticulum stress and causes pancreatic beta-cell dysfunction and apoptosis. J Biol Chem 282:3989-3997
Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR (1982) Analysis of nitrate, nitrite, and nitrate in biological fluids. Anal Biochem 126:131-138
Jobin C, Panja A, Hellerbrand C et al (1998) Inhibition of proinflammatory molecule production by adenovirus-mediated expression of a nuclear factor κB super-repressor in human intestinal epithelial cells. J Immunol 160:410-418
Hoorens A, Stange G, Pavlovic D, Pipeleers D (2001) Distinction between interleukin-1-induced necrosis and apoptosis of islet cells. Diabetes 50:551-557
Magnusson NE, Cardozo AK, Kruhoffer M, Eizirik DL, Orntoft TF, Jensen JL (2005) Construction and validation of the APOCHIP, a spotted oligo-microarray for the study of beta-cell apoptosis. BMC Bioinformatics 6:311
Pirot P, Naamane N, Libert F et al (2007) Global profiling of genes modified by endoplasmic reticulum stress in pancreatic beta cells reveals the early degradation of insulin mRNAs. Diabetologia 50:1006-1014
Darville MI, Eizirik DL (1998) Regulation by cytokines of the inducible nitric oxide synthase promoter in insulin-producing cells. Diabetologia 41:1101-1108
Dooley TP, Curto EV, Davis RL, Grammatico P, Robinson ES, Wilborn TW (2003) DNA microarrays and likelihood ratio bioinformatic methods: discovery of human melanocyte biomarkers. Pigment Cell Res 16:245-253
Liuwantara D, Elliot M, Smith MW et al (2006) Nuclear factor-κB regulates beta-cell death: a critical role for A20 in beta-cell protection. Diabetes 55:2491-2501
Mandrup-Poulsen T, Bendtzen K, Dinarello CA, Nerup J (1987) Human tumor necrosis factor potentiates human interleukin 1-mediated rat pancreatic beta-cell cytotoxicity. J Immunol 139:4077-4082
Flodstrom M, Welsh N, Eizirik DL (1996) Cytokines activate the nuclear factor kappa B (NF-kappa B) and induce nitric oxide production in human pancreatic islets. FEBS Lett 385:4-6
Liu D, Pavlovic D, Chen MC, Flodstrom M, Sandler S, Eizirik DL (2000) Cytokines induce apoptosis in beta-cells isolated from mice lacking the inducible isoform of nitric oxide synthase (iNOS-/-). Diabetes 49:1116-1122
Naamane N, van Helden J, Eizirik DL (2007) In silico identification of NF-κB-regulated genes in pancreatic beta-cells. BMC Bioinformatics 8:55
Hammar EB, Irminger JC, Rickenbach K et al (2005) Activation of NF-κB by extracellular matrix is involved in spreading and glucose-stimulated insulin secretion of pancreatic beta cells. J Biol Chem 280:30630-30637
Connelly L, Palacios-Callender M, Ameixa C, Moncada S, Hobbs AJ (2001) Biphasic regulation of NF-κB activity underlies the pro- and anti-inflammatory actions of nitric oxide. J Immunol 166:3873-3881
Nasuhara Y, Adcock IM, Catley M, Barnes PJ, Newton R (1999) Differential IκB kinase activation and IκBa degradation by interleukin-1β and tumor necrosis factor-α in human U937 monocytic cells. Evidence for additional regulatory steps in κB-dependent transcription. J Biol Chem 274:19965-19972
Chen LF, Greene WC (2004) Shaping the nuclear action of NF-κB. Nat Rev Mol Cell Biol 5:392-401
Pavlovic D, Andersen NA, Mandrup-Poulsen T, Eizirik DL (2000) Activation of extracellular signal-regulated kinase (ERK)1/2 contributes to cytokine-induced apoptosis in purified rat pancreatic beta-cells. Eur Cytokine Netw 11:267-274
Kim HS, Kim S, Lee MS (2005) IFN-γ sensitizes MIN6N8 insulinoma cells to TNF-α-induced apoptosis by inhibiting NF-κB-mediated XIAP upregulation. Biochem Biophys Res Commun 336:847-853
Kharroubi I, Lee CH, Hekerman P, Darville MI, Evans RM, Eizirik DL et al (2006) BCL-6: a possible missing link for anti-inflammatory PPAR-δ signalling in pancreatic beta cells. Diabetologia 49:2350-2358
