Proteomics Analysis of Cytokine-induced Dysfunction and Death in Insulin-producing INS-1E Cells

D’Hertog, Wannes; Overbergh, Lut; Lage, Kasper; Ferreira, Gabriela B; Maris, Michael; Gysemans, Conny; Flamez, Daisy; Cardozo, Alessandra K; Bergh, Gert Van den; Schoofs, Liliane; Arckens, Lutgarde; Moreau, Yves; Hansen, Daniel; Eizirik, Décio L.; Waelkens, Ettienne; Mathieu, Chantal

doi:10.1074/mcp.m700085-mcp200

Cited by 77 publications

(123 citation statements)

References 61 publications

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“…Protein identification Spots were picked from preparative gels with 350 μg protein lysate and trypsin digested as described previously [20]. MS analysis was performed by 4800 MALDI-TOF/TOF (Applied Biosystems, Carlsbad, CA, USA) and individual peptides from the MS/MS analysis were manually filtered; those with an individual expected value >0.05 were deleted, as were identifications based on a single peptide.…”

Section: Methodsmentioning

confidence: 99%

Early differences in islets from prediabetic NOD mice: combined microarray and proteomic analysis

et al. 2017

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Aims/hypothesis Type 1 diabetes is an endocrine disease where a long preclinical phase, characterised by immune cell infiltration in the islets of Langerhans, precedes elevated blood glucose levels and disease onset. Although several studies have investigated the role of the immune system in this process of insulitis, the importance of the beta cells themselves in the initiation of type 1 diabetes is less well understood. The aim of this study was to investigate intrinsic differences present in the islets from diabetes-prone NOD mice before the onset of insulitis. Methods The islet transcriptome and proteome of 2-3-week-old mice was investigated by microarray and 2-dimensional difference gel electrophoresis (2D-DIGE), respectively. Subsequent analyses using sophisticated pathway analysis and ranking of differentially expressed genes and proteins based on their relevance in type 1 diabetes were performed. Results In the preinsulitic period, alterations in general pathways related to metabolism and cell communication were already present. Additionally, our analyses pointed to an important role for post-translational modifications (PTMs), especially citrullination by PAD2 and protein misfolding due to low expression levels of protein disulphide isomerases (PDIA3, 4 and 6), as causative mechanisms that induce beta cell stress and potential auto-antigen generation. Conclusions/interpretation We conclude that the pancreatic islets, irrespective of immune differences, may contribute to the initiation of the autoimmune process. Data availability All microarray data are available in the ArrayExpress database (www.ebi.ac.uk/arrayexpress) under accession number E-MTAB-5264.

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Section: Methodsmentioning

confidence: 99%

Early differences in islets from prediabetic NOD mice: combined microarray and proteomic analysis

et al. 2017

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“…To investigate the underlying molecular pathways associated with the observed effects of GLP-1 on β-cell dysfunction and destruction, we performed a full proteome and protein interaction network analysis. Because 72 h of cytokine treatment resulted in only 18 ± 3.5% apoptotic β-cells, we decided to maintain these culture conditions for proteomic analysis, which thereby allowed us to identify networks of 17 The use of whole human islets enabled us to confirm different important pathways that are involved in cytokine-mediated β-cell destruction, extending our previous findings in rodent-derived INS-1E cells toward a more physiological model. 17 GLP-1 by itself affected the proteome profile of human islets only in a minor way, with only 10 proteins altered versus unexposed islets.…”

Section: ■ Discussionmentioning

confidence: 68%

“…Because 72 h of cytokine treatment resulted in only 18 ± 3.5% apoptotic β-cells, we decided to maintain these culture conditions for proteomic analysis, which thereby allowed us to identify networks of 17 The use of whole human islets enabled us to confirm different important pathways that are involved in cytokine-mediated β-cell destruction, extending our previous findings in rodent-derived INS-1E cells toward a more physiological model. 17 GLP-1 by itself affected the proteome profile of human islets only in a minor way, with only 10 proteins altered versus unexposed islets. However, when human islets were treated with the cytokine mixture IL-1β and IFN-γ, inducing increased apoptosis and decreased GSIS, a clear cytoprotective effect of GLP-1 was observed upon coincubation, correlating with published in vitro studies in human islets 14 and in vivo studies in murine models.…”

Section: ■ Discussionmentioning

confidence: 68%

“…45,46 In addition, we already observed and described the effect of a 24 h cytokine treatment on the reorganization of filamentous actin (F-actin) in rodent INS-1E cells. 17 The dynamic assembly and disassembly of filaments and the formation of larger scale filament structures, F-actin out of monomeric G-actin, are crucial aspects of the function of actin and are therefore under scrupulous control by over 100 actinbinding proteins. 47 In this study, we identified multiple cytokine-and GLP-1-responsive proteins such as actin-related protein 3 (ARP3), F-actin-capping protein subunit beta (CAPZB), and heat shock protein beta-1 (HSPB1), which are involved in F-actin polymerization 48,49 as well as proteins such as tropomodulin-3 (TMOD3), which is involved in actin depolymerization.…”

Section: Journal Of Proteome Researchmentioning

confidence: 99%

“…16 The molecular mechanisms by which cytokines affect β-cell function and survival and by which GLP-1 may protect against this inflammatory attack are still marginally understood. Here, as a follow-up of previous studies by D'Hertog et al 17 and Tews et al, 18 proteome analysis of human islets was performed using 2D difference gel electrophoresis (2D-DIGE) combined with protein interaction network analysis. This increases our understanding of the mechanisms involved in these processes because the current knowledge is largely based on RNA-based studies, 19−25 while many forms of regulation take place at a post-transcriptional level, such as post-translational modifications (PTMs) and proteolytic degradation.…”

Section: ■ Introductionmentioning

confidence: 99%

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Glucagon-Like Peptide-1 Protects Human Islets against Cytokine-Mediated β-Cell Dysfunction and Death: A Proteomic Study of the Pathways Involved

Rondas¹,

Bugliani

D’Hertog³

et al. 2013

J. Proteome Res.

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Glucagon-like peptide-1 (GLP-1) has been shown to protect pancreatic β-cells against cytokine-induced dysfunction and destruction. The mechanisms through which GLP-1 exerts its effects are complex and still poorly understood. The aim of this study was to analyze the protein expression profiles of human islets of Langerhans treated with cytokines (IL-1β and IFN-γ) in the presence or absence of GLP-1 by 2D difference gel electrophoresis and subsequent protein interaction network analysis to understand the molecular pathways involved in GLP-1-mediated β-cell protection. Co-incubation of cytokine-treated human islets with GLP-1 resulted in a marked protection of β-cells against cytokine-induced apoptosis and significantly attenuated cytokine-mediated inhibition of glucose-stimulated insulin secretion. The cytoprotective effects of GLP-1 coincided with substantial alterations in the protein expression profile of cytokine-treated human islets, illustrating a counteracting effect on proteins from different functional classes such as actin cytoskeleton, chaperones, metabolic proteins, and islet regenerating proteins. In summary, GLP-1 alters in an integrated manner protein networks in cytokine-exposed human islets while protecting them against cytokine-mediated cell death and dysfunction. These data illustrate the beneficial effects of GLP-1 on human islets under immune attack, leading to a better understanding of the underlying mechanisms involved, a prerequisite for improving therapies for diabetic patients.

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Genetic tools and algorithms for gene discovery in major congenital anomalies

Donahoe¹,

Noonan²,

Lage³

2008

Birth Defects Research

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Proteomics Analysis of Cytokine-induced Dysfunction and Death in Insulin-producing INS-1E Cells

Cited by 77 publications

References 61 publications

Early differences in islets from prediabetic NOD mice: combined microarray and proteomic analysis

Early differences in islets from prediabetic NOD mice: combined microarray and proteomic analysis

Glucagon-Like Peptide-1 Protects Human Islets against Cytokine-Mediated β-Cell Dysfunction and Death: A Proteomic Study of the Pathways Involved

Genetic tools and algorithms for gene discovery in major congenital anomalies

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