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 9 publications

(12 citation statements)

References 65 publications

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“…1I). To determine if cytokines decreased LGR4 expression specifically in β-cells, we analyzed the published transcriptome of control (vehicle-treated) and cytokine-treated human EndoC-βH5 β-cell line (30) and found a significant decrease in LGR4 mRNA levels 48h post cytokine-treatment (Fig. 1J).…”

Section: Resultsmentioning

confidence: 99%

“…(I) qPCR analysis of LGR4 expression relative to BETA ACTIN in an independent cohort of human islets treated with vehicle (CTRL) or cytokine mix (CYT) for 24h (n=4). (J) LGR4 expression in the human β-cell line (EndoC-βH5) treated with vehicle (CTRL) or CYT for 48h based on meta-analysis of published transcriptome data (30) using https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE218735 (n=4). *p<0.05, **p<0.01, ***p<0.001 vs 4 month or CTRL.…”

Section: Resultsmentioning

confidence: 99%

“…An independent set of human islet preps (Supplemental Table 1) were used to confirm decrease in LGR4 mRNA under cytokine treatment vs. control by qPCR. Data for LGR4 mRNA levels in EndoC-βH5 human β cell line was retrieved from bulk RNA analysis published by Frørup et al (30), available at: https://www.ncbi.nlm.nih.gov/geo/geo2r/?acc=GSE218735. LGR4 expression values obtained under cytokine treatment for each experiment were normalized to vehicle control and expressed as average fold change.…”

Section: Methodsmentioning

confidence: 99%

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LGR4 is essential for maintaining β-cell homeostasis through suppression of RANK

Filipowska,

Cisneros,

Leon-Rivera

et al. 2024

Preprint

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Pancreatic beta-cell stress contributes to diabetes progression. This study demonstrates that Leucine-rich repeat-containing G-protein-coupled-receptor-4 (LGR4) is critical for maintaining beta-cell health and is modulated by stressors. In vitro, Lgr4 knockdown decreases proliferation and survival in rodent beta-cells, while overexpression protects against cytokine-induced cell death in rodent and human beta-cells. Mechanistically, LGR4 suppresses Receptor Activator of Nuclear Factor Kappa B (NFkappaB) (RANK) and its subsequent activation of NFkappaB to protect beta-cells. Beta-cell-specific Lgr4-conditional knockout (cko) mice exhibit normal glucose homeostasis but increased beta-cell death in both sexes and decreased proliferation only in females. Male Lgr4cko mice under stress display reduced beta-cell proliferation and a further increase in beta-cell death. Upon aging, both male and female Lgr4cko mice display impaired beta-cell homeostasis, however, only female mice are glucose intolerant with decreased plasma insulin. We show that LGR4 is required for maintaining beta-cell health under basal and stress-induced conditions, through suppression of RANK.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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LGR4 is essential for maintaining β-cell homeostasis through suppression of RANK

Filipowska,

Cisneros,

Leon-Rivera

et al. 2024

Preprint

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“…Cytokines can control the recruitment of immune cells. As an example, oral Lactococcus secreted IL10 to limit islet infiltration by immune cells and prevent and halt the progression of diabetes [ 67 , 68 , 69 ]. Similarly, our oral Salmonella vaccine induced tolerance, in part, by elevating IL10, IL2, IL13, and MCP-1 and lowering IFNγ, GM-CSF, TNFα, IL7, and IL12, in responder animals.…”

Section: Discussionmentioning

confidence: 99%

Mechanism of Action of Oral Salmonella-Based Vaccine to Prevent and Reverse Type 1 Diabetes in NOD Mice

Cobb,

Rawson,

Gonzalez

et al. 2024

Vaccines

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A combination therapy of preproinsulin (PPI) and immunomodulators (TGFβ+IL10) orally delivered via genetically modified Salmonella and anti-CD3 promoted glucose balance in in NOD mice with recent onset diabetes. The Salmonella bacteria were modified to express the diabetes-associated antigen PPI controlled by a bacterial promoter in conjunction with over-expressed immunomodulating molecules. The possible mechanisms of action of this vaccine to limit autoimmune diabetes remained undefined. In mice, the vaccine prevented and reversed ongoing diabetes. The vaccine-mediated beneficial effects were associated with increased numbers of antigen-specific CD4+CD25+Foxp3+ Tregs, CD4+CD49b+LAG3+ Tr1-cells, and tolerogenic dendritic-cells (tol-DCs) in the spleens and lymphatic organs of treated mice. Despite this, the immune response to Salmonella infection was not altered. Furthermore, the vaccine effects were associated with a reduction in islet-infiltrating lymphocytes and an increase in the islet beta-cell mass. This was associated with increased serum levels of the tolerogenic cytokines (IL10, IL2, and IL13) and chemokine ligand 2 (CCL2) and decreased levels of inflammatory cytokines (IFNγ, GM-CSF, IL6, IL12, and TNFα) and chemokines (CXCL1, CXCL2, and CXCL5). Overall, the data suggest that the Salmonella-based vaccine modulates the immune response, reduces inflammation, and promotes tolerance specifically to an antigen involved in autoimmune diabetes.

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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

Self Cite

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

Cited by 9 publications

References 65 publications

LGR4 is essential for maintaining β-cell homeostasis through suppression of RANK

LGR4 is essential for maintaining β-cell homeostasis through suppression of RANK

Mechanism of Action of Oral Salmonella-Based Vaccine to Prevent and Reverse Type 1 Diabetes in NOD Mice

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

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