Inhibition of Cytokine-Induced NF-κB Activation by Adenovirus-Mediated Expression of a NF-κB Super-Repressor Prevents β-Cell Apoptosis

Heimberg, Harry; Heremans, Yves; Jobin, Christian; Leemans, Ruth; Cardozo, Alessandra K; Darville, Martine I.; Eizirik, Décio L.

doi:10.2337/diabetes.50.10.2219

Cited by 254 publications

(248 citation statements)

References 40 publications

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“…It has been suggested by others that GFP is itself intrinsically toxic towards rat beta cells and other cell types [27,28]. In this study, there was no evidence of increased apoptosis or necrosis of cells expressing GFP, regardless of whether expression was driven by the insulin or CMV promoter.…”

Section: Discussionsupporting

confidence: 40%

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

et al. 2002

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Aims/hypothesis. Human islet cells survive poorly in culture and are overgrown by non-endocrine cells. The aims of this study were to sort human beta cells and to develop approaches for their improved survival in culture. Methods. Human islets were infected with recombinant adenovirus expressing green fluorescent protein (GFP) under the control of the rat insulin promoter such that only beta cells expressed GFP. GFP-positive beta cells were sorted by flow cytometry, and expression of select integrins evaluated by RT-PCR. Beta cells were cultured on different extracellular matrices for up to 15 days. Apoptosis was measured by annexin V binding and ELISA. Insulin secretion was measured by ELISA. Results. Sorted beta cells survived less well in culture than unsorted islet cells. This did not appear to be due to adenoviral infection and/or GFP expression. Purified beta cells expressed the integrins α3, α5, α6, αV, β1, but not β4. Of the various matrices tested, sorted beta cells attached and spread best on a lawn of lysed human bladder carcinoma cells (5637 cells). However, survival remained poor. Cell death was decreased but not prevented by continued presence of 10 mmol/l nicotinamide and apoptosis decreased by 24 h incubation with 20 µmol/l Z-VAD. Insulin secretion was maintained over 6 days following treatment with both agents. Conclusions/interpretation. Purification of human beta cells induces marked apoptosis limiting their function and survival in vitro. This was improved by matching the extracellular matrix to the specific expression of integrins and by addition of nicotinamide and Z-VAD. [Diabetologia (2002) 45:841-850]

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Section: Discussionsupporting

confidence: 40%

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

et al. 2002

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“…Adenoviruses were generated using the pShuttle/pAdEasy-1 vector system [17,21]. Adenoviruses expressing the enhanced green fluorescent protein (Ad-eGFP) or adenoviruses expressing the luciferase protein (Ad-LUC) were used as controls [22]. Rat dispersed islets/FACS-purified beta cells were infected as previously described [22].…”

Section: Methodsmentioning

confidence: 99%

Sustained production of spliced X-box binding protein 1 (XBP1) induces pancreatic beta cell dysfunction and apoptosis

et al. 2010

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Aims/hypothesis Pro-inflammatory cytokines involved in the pathogenesis of type 1 diabetes deplete endoplasmic reticulum (ER) Ca 2+ stores, leading to ER-stress and beta cell apoptosis. However, the cytokine-induced ER-stress response in beta cells is atypical and characterised by induction of the pro-apoptotic PKR-like ER kinase (PERK)-C/EBP homologous protein (CHOP) branch of the unfolded protein response, but defective X-box binding protein 1 (XBP1) splicing and activating transcription factor 6 activation. The purpose of this study was to overexpress spliced/active Xbp1 (XBP1s) to increase beta cell resistance to cytokine-induced ER-stress and apoptosis.Methods Xbp1s was overexpressed using adenoviruses and knocked down using small interference RNA in rat islet cells. In selected experiments, Xbp1 was also knocked down in FACS-purified rat beta cells and rat fibroblasts. Expression and production of XBP1s and key downstream genes and proteins was measured and beta cell function and viability were evaluated.

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“…NFκB is a point of convergence in the signal transduction pathways activated by the proinflammatory cytokine IL-1β in beta cells, and plays a pivotal role in the cytokineinduced beta cell death seen in type 1 diabetes [15][16][17]. NFκB controls induction of several inflammatory genes, one being the gene encoding inducible NO synthase (iNOS), leading to NO formation and beta cell death [17,18].…”

Section: Introductionmentioning

confidence: 99%

“…NFκB controls induction of several inflammatory genes, one being the gene encoding inducible NO synthase (iNOS), leading to NO formation and beta cell death [17,18]. Blockade of the NFκB pathway has been shown to inhibit Fas-triggered apoptosis and cytokine-induced suppression of glucose-stimulated insulin secretion in human islets [16], as well as expression of the nitric oxide synthase 2 gene, NO formation and cell death by apoptosis and necrosis in primary rat islets [17].…”

Section: Introductionmentioning

confidence: 99%

Inhibition of histone deacetylases prevents cytokine-induced toxicity in beta cells

et al. 2007

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Aims/hypothesis The immune-mediated elimination of pancreatic beta cells in type 1 diabetes involves release of cytotoxic cytokines such as IL-1β and IFNγ, which induce beta cell death in vitro by mechanisms that are both dependent and independent of nitric oxide (NO). Nuclear factor kappa B (NFκB) is a critical signalling molecule in inflammation and is required for expression of the gene encoding inducible NO synthase (iNOS) and of proapoptotic genes. NFκB has recently been shown to associate with chromatin-modifying enzymes histone acetyltransferases and histone deacetylases (HDAC), and positive effects of HDAC inhibition have been obtained in several inflammatory diseases. Thus, the aim of this study was to investigate whether HDAC inhibition protects beta cells against cytokine-induced toxicity. Materials and methods The beta cell line, INS-1, or intact rat islets were precultured with HDAC inhibitors suberoylanilide hydroxamic acid or trichostatin A in the absence or presence of IL-1β and IFNγ. Effects on insulin secretion and NO formation were measured by ELISA and Griess reagent, respectively. iNOS levels and NFκB activity were measured by immunoblotting and by immunoblotting combined with electrophoretic mobility shift assay, respectively. Viability was analysed by 3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl-tetrazolium bromide and apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) assay and histone-DNA complex ELISA. Results HDAC inhibition reduced cytokine-mediated decrease in insulin secretion and increase in iNOS levels, NO formation and apoptosis. IL-1β induced a bi-phasic phosphorylation of inhibitor protein kappa Bα (IκBα) with the 2nd peak being sensitive to HDAC inhibition. No effect was seen on IκBα degradation and NFκB DNA binding. Conclusions/interpretation HDAC inhibition prevents cytokine-induced beta cell apoptosis and impaired beta cell function associated with a downregulation of NFκB transactivating activity.

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Inhibition of Cytokine-Induced NF-κB Activation by Adenovirus-Mediated Expression of a NF-κB Super-Repressor Prevents β-Cell Apoptosis

Cited by 254 publications

References 40 publications

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

Impact of integrin-matrix matching and inhibition of apoptosis on the survival of purified human beta-cells in vitro

Sustained production of spliced X-box binding protein 1 (XBP1) induces pancreatic beta cell dysfunction and apoptosis

Inhibition of histone deacetylases prevents cytokine-induced toxicity in beta cells

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