Research-Focused Isolation of Human Islets From Donors With and Without Diabetes at the Alberta Diabetes Institute IsletCore

Lyon, James; Fox, Jocelyn E. Manning; Spigelman, Aliya F; Kim, Ryekjang; Smith, Nancy; O’Gorman, Doug; Kin, Tatsuya; Shapiro, A. M. James; Rajotte, R. V.; MacDonald, Patrick E.

doi:10.1210/en.2015-1562

Cited by 120 publications

(155 citation statements)

References 45 publications

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“…A subset of human islets (n = 20 donors) was obtained from the Alberta Diabetes Institute IsletCore [37] at the University of Alberta. The remainder of donor islets (n = 24 donors) was obtained from islet isolation centers participating in the Integrated Islet Distribution Program (http://iidp.coh.org/).…”

Section: Methodsmentioning

confidence: 99%

Opposing effects of prostaglandin E 2 receptors EP3 and EP4 on mouse and human β-cell survival and proliferation

Carboneau

Allan

Townsend

et al. 2017

Molecular Metabolism

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ObjectiveHyperglycemia and systemic inflammation, hallmarks of Type 2 Diabetes (T2D), can induce the production of the inflammatory signaling molecule Prostaglandin E2 (PGE2) in islets. The effects of PGE2 are mediated by its four receptors, E-Prostanoid Receptors 1-4 (EP1-4). EP3 and EP4 play opposing roles in many cell types due to signaling through different G proteins, Gi and GS, respectively. We previously found that EP3 and EP4 expression are reciprocally regulated by activation of the FoxM1 transcription factor, which promotes β-cell proliferation and survival. Our goal was to determine if EP3 and EP4 regulate β-cell proliferation and survival and, if so, to elucidate the downstream signaling mechanisms.Methodsβ-cell proliferation was assessed in mouse and human islets ex vivo treated with selective agonists and antagonists for EP3 (sulprostone and DG-041, respectively) and EP4 (CAY10598 and L-161,982, respectively). β-cell survival was measured in mouse and human islets treated with the EP3- and EP4-selective ligands in conjunction with a cytokine cocktail to induce cell death. Changes in gene expression and protein phosphorylation were analyzed in response to modulation of EP3 and EP4 activity in mouse islets.ResultsBlockade of EP3 enhanced β-cell proliferation in young, but not old, mouse islets in part through phospholipase C (PLC)-γ1 activity. Blocking EP3 also increased human β-cell proliferation. EP4 modulation had no effect on ex vivo proliferation alone. However, blockade of EP3 in combination with activation of EP4 enhanced human, but not mouse, β-cell proliferation. In both mouse and human islets, EP3 blockade or EP4 activation enhanced β-cell survival in the presence of cytokines. EP4 acts in a protein kinase A (PKA)-dependent manner to increase mouse β-cell survival. In addition, the positive effects of FoxM1 activation on β-cell survival are inhibited by EP3 and dependent on EP4 signaling.ConclusionsOur results identify EP3 and EP4 as novel regulators of β-cell proliferation and survival in mouse and human islets ex vivo.

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Section: Methodsmentioning

confidence: 99%

Opposing effects of prostaglandin E 2 receptors EP3 and EP4 on mouse and human β-cell survival and proliferation

Carboneau

Allan

Townsend

et al. 2017

Molecular Metabolism

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“…Islets from 50 human donors were isolated by the Alberta Diabetes Institute IsletCore (22) or the Clinical Islet Laboratory (23) at the University of Alberta, with appropriate ethical approval from the University of Alberta Human Research Ethics Board (Pro00013094; Pro 00001754). Donor information is described in Supplementary Tables 1 and 2.…”

Section: Cell Culture and Transfectionmentioning

confidence: 99%

A Glycine-Insulin Autocrine Feedback Loop Enhances Insulin Secretion From Human β-Cells and Is Impaired in Type 2 Diabetes

et al. 2016

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The secretion of insulin from pancreatic islet β-cells is critical for glucose homeostasis. Disrupted insulin secretion underlies almost all forms of diabetes, including the most common form, type 2 diabetes (T2D). The control of insulin secretion is complex and affected by circulating nutrients, neuronal inputs, and local signaling. In the current study, we examined the contribution of glycine, an amino acid and neurotransmitter that activates ligand-gated Cl− currents, to insulin secretion from islets of human donors with and without T2D. We find that human islet β-cells express glycine receptors (GlyR), notably the GlyRα1 subunit, and the glycine transporter (GlyT) isoforms GlyT1 and GlyT2. β-Cells exhibit significant glycine-induced Cl− currents that promote membrane depolarization, Ca2+ entry, and insulin secretion from β-cells from donors without T2D. However, GlyRα1 expression and glycine-induced currents are reduced in β-cells from donors with T2D. Glycine is actively cleared by the GlyT expressed within β-cells, which store and release glycine that acts in an autocrine manner. Finally, a significant positive relationship exists between insulin and GlyR, because insulin enhances the glycine-activated current in a phosphoinositide 3-kinase–dependent manner, a positive feedback loop that we find is completely lost in β-cells from donors with T2D.

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“…Human islets were isolated from donor organs at the Alberta Diabetes Institute IsletCore (www.bcell.org/IsletCore.html) or the Clinical Islet Laboratory at the University of Alberta as described [29] and cultured as described [30]. Information on donor age, BMI, sex and HbA1c were provided.…”

Section: Methodsmentioning

confidence: 99%

STEAP4 expression in human islets is associated with differences in body mass index, sex, HbA1c, and inflammation

et al. 2017

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Six-transmembrane epithelial antigen of the prostate 4 (STEAP4) is a metalloreductase that has been shown previously to protect cells from inflammatory damage. Genetic variants in STEAP4 have been associated with numerous metabolic disorders related to obesity, including putative defects in the acute insulin response to glucose in type 2 diabetes (T2D). Purpose We examined whether obesity and/or T2D altered STEAP4 expression in human pancreatic islets. Methods Human islets were isolated from deceased donors at two medical centers and processed for quantitative PCR. Organ donors were selected by status as non-diabetic or having T2D. Site1 (Edmonton): N=13 T2D donors (7M, 6F), N=20 non-diabetic donors (7M, 13F). Site2 (Virginia): N=6 T2D donors (6F), N=6 non-diabetic donors (3M, 3F). Results STEAP4 showed reduced islet expression with increasing BMI among all donors (P<0.10) and non-diabetic donors (P<0.05) from Site1; STEAP4 showed reduced islet expression among T2D donors with increasing HbA1c. Islet STEAP4 expression was also marginally higher in female donors (p<0.10). Among T2D donors from Site2, islet insulin expression was reduced, STEAP4 expression was increased, and white blood cell counts were increased compared to non-diabetic donors. Islets from non-diabetic donors that were exposed overnight to 5ng/ml IL-1β displayed increased STEAP4 expression, consistent with STEAP4 upregulation by inflammatory signaling. Conclusions These findings suggest that increased STEAP4 mRNA expression is associated with inflammatory stimuli, whereas lower STEAP4 expression is associated with obesity in human islets. Given its known protective role, downregulation of STEAP4 by chronic obesity suggests a mechanism for reduced islet protection against cellular damage.

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Research-Focused Isolation of Human Islets From Donors With and Without Diabetes at the Alberta Diabetes Institute IsletCore

Cited by 120 publications

References 45 publications

Opposing effects of prostaglandin E 2 receptors EP3 and EP4 on mouse and human β-cell survival and proliferation

Opposing effects of prostaglandin E 2 receptors EP3 and EP4 on mouse and human β-cell survival and proliferation

A Glycine-Insulin Autocrine Feedback Loop Enhances Insulin Secretion From Human β-Cells and Is Impaired in Type 2 Diabetes

STEAP4 expression in human islets is associated with differences in body mass index, sex, HbA1c, and inflammation

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