Characterization of the functional and transcriptomic effects of pro-inflammatory cytokines on human EndoC-βH5 beta cells

Frørup, Caroline; Gerwig, Rebekka; Svane, Cecilie A. S.; Melo, Joana Mendes Lopes de; Henriksen, Kristine; Fløyel, Tina; Pociot, Flemming; Kaur, Simranjeet; Størling, Joachim

doi:10.3389/fendo.2023.1128523

Cited by 5 publications

(1 citation statement)

References 65 publications

(114 reference statements)

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“…This may reflect different roles of gasdermins in beta-cell inflammation during development of T1D and during sustained inflammation in type 2 diabetes versus established T1D where most of the beta cells have perished. We also examined if pro-inflammatory cytokines, known as key drivers of beta-cell damage and death, upregulate pyroptosis genes in human islets and human EndoC-βH5 beta cells [54]. The heatmaps in Figure 1B and C generated from in-house RNA sequencing data show that cytokines increase the expression of several pyroptosis genes in human islets and EndoC-βH5 cells.…”

Section: Introductionmentioning

confidence: 99%

Beta-Cell Pyroptosis – A Burning Flame in Type 1 Diabetes?

Frørup,

Svane,

Henriksen

et al. 2024

Preprint

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Immune-mediated destruction of the beta-cells in the pancreatic islets of Langerhans is the underlying cause of type 1 diabetes (T1D). Despite decades of research, the exact mechanisms involved at the beta-cell level during the development of disease remain poorly understood. This includes the mode(s) of beta-cell death and signaling events implicated in exacerbating local islet inflammation and immune cell infiltration, commonly known as insulitis. In disease models, beta-cell apoptosis seems to be the predominant cell death form which has led to the general assumption that beta cells mostly die by apoptosis in T1D. However, apoptosis is an anti-inflammatory programmed cell death mechanism, and this dogma is therefore challenged by the pathogenetic nature of T1D, as a progressive increase in islet inflammation is seen. This infers that other modes of beta-cell death that inherently increase insulitis may predominate. One such mechanism could be the newly characterized form of programmed cell death; pyroptosis (from the Greek 'fire-falling'). Pyroptosis is characterized by gasdermin-mediated cell lysis with a bursting release of pro-inflammatory factors. Beta-cell death by pyroptosis in T1D may therefore offer a plausible explanation for the exacerbated paracrine islet inflammation that spreads during the progression of insulitis. Here, we briefly debate the evidence supporting beta-cell pyroptosis in T1D as a central mechanism of islet inflammation and beta-cell demise. The paper intends to challenge the current understanding of beta-cell destruction to move the field forward. Importantly, we present experimental data from human islets and EndoC-βH5 cells that directly support beta-cell pyroptosis as a rational death mechanism in T1D. We suggest a model of beta-cell demise in T1D in which pyroptosis plays a prominent role in concert with other cell death mechanisms. As the role of pyroptosis in disease is still in its infancy, we hope also to inspire researchers working in other disease fields.

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Section: Introductionmentioning

confidence: 99%

Beta-Cell Pyroptosis – A Burning Flame in Type 1 Diabetes?

Frørup,

Svane,

Henriksen

et al. 2024

Preprint

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EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion

Blanchi,

Taurand,

Colace

et al. 2023

Molecular Metabolism

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Renewable Human Cell Model for Type 1 Diabetes Research: EndoC-βH5/HUVEC Coculture Spheroids

Porter,

Yitayew,

Tabrizian

2023

Journal of Diabetes Research

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In vitro drug screening for type 1 diabetes therapies has largely been conducted on human organ donor islets for proof of efficacy. While native islets are the ultimate target of these drugs (either in situ or for transplantation), significant benefit can be difficult to ascertain due to the highly heterogeneous nature of individual donors and the overall scarcity of human islets for research. We present an in vitro coculture model based on immortalized insulin-producing beta-cell lines with human endothelial cells in 3D spheroids that aims to recapitulate the islet morphology in an effort towards developing a standardized cell model for in vitro diabetes research. Human insulin-producing immortalized EndoC-βH5 cells are cocultured with human endothelial cells in varying ratios to evaluate 3D cell culture models for type 1 diabetes research. Insulin secretion, metabolic activity, live cell fluorescence staining, and gene expression assays were used to compare the viability and functionality of spheroids composed of 100% beta-cells, 1 : 1 beta-cell/endothelial, and 1 : 3 beta-cell/endothelial. Monoculture and βH5/HUVEC cocultures formed compact spheroids within 7 days, with average diameter ~140 μm. This pilot study indicated that stimulated insulin release from 0 to 20 mM glucose increased from ~8-fold for monoculture and 1 : 1 coculture spheroids to over 20-fold for 1 : 3 EndoC-βH5/HUVEC spheroids. Metabolic activity was also ~12% higher in the 1 : 3 EndoC-βH5/HUVEC group compared to other groups. Stimulating monoculture beta-cell spheroids with 20 mM glucose +1 μg/mL glycine-modified INGAP-P increased the insulin stimulation index ~2-fold compared to glucose alone. Considering their availability and consistent phenotype, EndoC-βH5-based spheroids present a useful 3D cell model for in vitro testing and drug screening applications.

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Characterization of the functional and transcriptomic effects of pro-inflammatory cytokines on human EndoC-βH5 beta cells

Cited by 5 publications

References 65 publications

Beta-Cell Pyroptosis – A Burning Flame in Type 1 Diabetes?

Beta-Cell Pyroptosis – A Burning Flame in Type 1 Diabetes?

EndoC-βH5 cells are storable and ready-to-use human pancreatic beta cells with physiological insulin secretion

Renewable Human Cell Model for Type 1 Diabetes Research: EndoC-βH5/HUVEC Coculture Spheroids

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