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. 2023 May 11;299(6):104816. doi: 10.1016/j.jbc.2023.104816

Figure 7.

Figure 7

CRN02481 significantly curtails insulin secretion in pancreatic islets from human healthy donors and patients with HI. Isolated human healthy, control islets (Prodo Labs, CA) were loaded in a perifusion system (Biorep, FL) and treated over time with 3 mM glucose (G3), 16.7 mM glucose (G16.7), and 16.7 mM glucose + 100 μM tolbutamide (G16.7 + Tolb) in the presence of absence of increasing concentrations of CRN02481 as described in material and methods. Insulin secretion was quantified using an ELISA assay (Mercodia, Uppsala, Sweden) and the dynamic stimulation index (dSI) was calculated as stimulated insulin levels/basal insulin levels. (A) Mean dSI ± SD (n = 2 technical replicates) from one representative donor. B, percent insulin secretion was calculated in each condition and compared to the vehicle group which was set to 100%. One-way ANOVA analysis with Dunnett’s multiple comparisons: G16.7 (F(3, 12) = 10.21, p = 0.98 for 0.01 μM, p = 0.04 for 0.1 μM, and p = 0.002 for 1 μM compared to vehicle); G16.7 + Tolb (F(3, 12) = 14.49, p = 0.77 for 0.01 μM, p = 0.01 for 0.1 μM, and p = 0.001 for 1 μM compared to vehicle). Data represent mean ± SD from 4 independent donors compared to vehicle. C, Perifusion of primary isolated BWS-HI human islets to assess insulin release with treatment of CRN02481 (500 nM) in response to glucose ramp (0–25 mM) and KCl (Two-way ANOVA with Tukey’s: (F(1, 95) = 146.3, p < 0.0001). D, percent insulin release for BWS-HI (Patient #1) human islet perifusion. E, Perifusion of primary isolated KATP-HI human islets (Patient #2) to assess insulin release with treatment of CRN02481 (500 nM) in response to AAM ramp (0–12 mM) and KCl (Two-way ANOVA with Tukey’s: (F(1, 95) = 119.4, p < 0.0001). F, percent insulin release for KATP-HI Patient #2 perifusion. G, Perifusion of primary isolated KATP-HI human islets (Patient #3) (Two-way ANOVA with Tukey’s: (F(1, 95) = 163.5, p < 0.0001). H, percent insulin release for KATP-HI perifusion (Patient #3). (Compared to vehicle: ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001). Data represent mean ± SD compared to vehicle.