Beta-cell maturation leads to in vitro sensitivity to cytotoxins.

Nielsen, Kristina Tomra; Karlsen, Allan E.; Deckert, Markus; Madsen, Ole; Serup, Palle; Mandrup-Poulsen, Thomas; Nerup, Jørn

doi:10.2337/diabetes.48.12.2324

Cited by 58 publications

(62 citation statements)

References 12 publications

(12 reference statements)

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“…Previous analysis of mRNA gene expression profiles by semi-quantitative multiplex RT-PCR of the two NHI-phenotypes used in this study, grown under the same conditions, showed that several genes associated with late stages of betacell maturation, such as PDX-1 were expressed in both phenotypes, whereas Nkx-6.1 was restricted to the beta-cell phenotype [20]. We showed that maturation into the beta-cell phenotype was accompanied by insulin gene expression and an acquired susceptibility to toxic effects of IL-1β, not seen in the pre-beta-cell NHI-glu phenotype [12]. Proteome analyses of the two phenotypes could therefore identify proteins and pathways important for beta-cell maturation and the acquired IL-1β sensitivity.…”

Section: Discussionmentioning

confidence: 86%

“…The increased pool of arginine could then serve as substrate for iNOS and result in increased production of NO [37]. However, our previous analyses have shown that despite the different sensitivity to IL-1β, no significant difference in IL-1β induced NO production could be detected between the two phenotypes [12]. Nevertheless, it has been shown that IL-1β induces the citrulline-NO cycle in beta cells, and that extracellular arginine or citrulline is required for NO production [38].…”

Section: Protein Synthesis Chaperones and Protein Foldingmentioning

confidence: 91%

“…The NHI-cell system [12] is based on the subclone NHI-glu derived from the glucagon-producing MSL-G2 culture [17]. Following in vivo passage by transplantation into syngeneic NEDH rats, the NHI-glu cells mature into insulinomas [10,18].…”

Section: Methodsmentioning

confidence: 99%

“…Here we address this hypothesis by using a glucagon producing pre-beta-cell cell line (NHI-glu), which following in vivo passage as a subcutaneous tumour in a rat, was re-established as an insulin producing beta-cell line (NHIins) [10,11]. Our previous analyses of these two phenotypes showed that this maturation process was accompanied by an acquired sensitivity to the toxic effects of IL-1β [12]. Hence, the aim of our study was to characterize by proteome analysis [13,14,15] changes in protein expression accompanying the final maturation from the glucagon positive and IL-1β resistant pre-beta-cell phenotype to the insulin positive and IL-1β sensitive beta-cell phenotype and to relate these data to our previous analysis of PiPP in IL-1β exposed rat islets.…”

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confidence: 99%

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Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1?

et al. 2004

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Aim/hypothesis. Type 1 diabetes mellitus (T1DM) is caused by specific destruction of the pancreatic beta cells in the islets of Langerhans. Increased sensitivity to cytokines, in particular to interleukin-1β (IL-1β) seems to be an acquired trait during beta-cell maturation. In response to cytokines both protective and deleterious mechanisms are induced in beta cells, and when the deleterious prevail, T1DM develops. The aims of this study were to identify perturbation in protein patterns (PiPP) associated with beta-cell maturation, and compare these changes to previous analyses of IL-1β exposed rat islets. For this purpose, proteome analyses were carried out using a cell-line, which matures from a glucagon-producing pre-beta-cell phenotype (NHI-glu) to an insulin-producing beta-cell phenotype (NHI-ins). We have previously shown that this maturation is accompanied by acquired sensitivity to the toxic effects of IL-1β.Methods. 2D-gel electrophoresis was used to separate the proteins and MALDI-MS and database searches were performed to identify the proteins. Results. During beta-cell maturation 135 protein spots out of 2239 detectable changed expression levels. Of these, 74 were down-regulated, 44 up-regulated, 16 were suppressed and 1 was expressed de novo. Using MALDI-MS, positive identification was obtained for 93 out of the 135 protein-spots revealing 97 different proteins. Of these, 22 proteins were in common with changes identified in previous proteome analysis of perturbation in protein pattern in IL-1β exposed rat islets. Several of the proteins were present in more than one spot suggesting post-translational modification. Conclusion/interpretation. Several proteins and protein modifications were identified that could be critically involved in beta-cell maturation, insulin-gene expression and the acquired IL-1β sensitivity. [Diabetologia (2004) A. E. Karlsen ( ✉ ), Steno Diabetes Center, Niels Steensensvej 2, 2820 Gentofte, Denmark E-mail: aek@novonordisk.com Abbreviations: T1DM, Type 1 diabetes mellitus; PiPP, perturbation in protein pattern; NO, nitric oxide; iNOS, inducible nitric oxide synthase; 2D-GE, 2 dimensional gel electrophoresis; IEF, isoelectric focusing; NEPHGE, non-equilibrium pH-gradient electrophoresis; WF, Wistar Furth; BB, Bio Breeding; PDX-1, pancreatic duodenum homeobox 1; ASS, argininosuccinate synthetase; HSP, heat shock protein; Picot, PKC-interacting cousin of thioredoxin; JNK, Jun N-terminal kinase; VDAC, voltage-dependent anion channel; GST, glutathione-Stransferase; Mw, molecular weight; pI, isoelectric point. Diabetologia (2004) 47:62-74 DOI 10.1007/s00125-003-1277 Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1β Autoimmune Type 1 diabetes mellitus (T1DM) is caused by specific destruction of the insulin-producing beta cells in the islets of Langerhans in the pancreas [1]. During this process the islets are infiltrated with macrophages and lymphocytes, and these cells release a mixture of cytokines, such as inte...

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Section: Discussionmentioning

confidence: 86%

Section: Protein Synthesis Chaperones and Protein Foldingmentioning

confidence: 91%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1?

et al. 2004

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“…Accumulated insulin in the incubation media was measured as previously [28,29], except that the enzyme substrate used here was the TMB + ready-to-use-substrate (Kem En Tec, Taastrup, Denmark).…”

Section: Methodsmentioning

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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Toxin‐based selection of insulin‐producing cells with improved defense properties for islet cell transplantation

Bloch

Vardi

2005

Diabetes Metabolism Res

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Insulin-producing pancreatic beta-cells are known to be extremely susceptible to destruction, primarily by autoimmune mechanisms, infectious agents, and by chemical toxins that cause overt type I diabetes. As development of highly protected insulin-producing cells would be important for successful cell therapy of diabetic patients, gene transfection technique was utilized by several investigators in order to improve the defense properties of transplanted cells. In this article, we summarize other approaches based on a selection strategy that has been developed in our laboratory and by other research groups that engineer pancreatic beta-cells to provide protection against diabetogenic toxins (streptozotocin and alloxan), oxidative stress and cytokines. Selection strategies based on acute repeated or long-term continuous treatment of cell lines with cytotoxic agents have resulted in the selection of highly resistant cell subpopulations. We discuss possible involvement of different expression of cytoprotective genes in the selection of cell subpopulations, which demonstrate a broad spectrum of resistance. Importantly, toxin-based selection did not impair functional activity of the cells as it was shown in vitro. In addition, selected cells preserved their improved metabolic characteristics following encapsulation in alginate and subsequent implantation in diabetic animals. Identifying the mechanisms through which cell defense properties act will help clarify the process responsible for beta-cell regeneration in type I diabetes patients. Such knowledge might be useful in developing strategies focusing on the regeneration of beta-cell resistant populations.

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Beta-cell maturation leads to in vitro sensitivity to cytotoxins.

Cited by 58 publications

References 12 publications

Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1?

Protein expression changes in a cell system of beta-cell maturation reflect an acquired sensitivity to IL-1?

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

Toxin‐based selection of insulin‐producing cells with improved defense properties for islet cell transplantation

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