Fig. 5: Repeat i.v. dose of hG6PC mRNA-LNP results in safe and effective restoration of euglycemia in L.G6pc^−/− mice.

a Single-dose duration of action of hG6PC S298C mRNA-LNP (0.5 or 1.0 mg/kg) administered i.v. in L.G6pc^−/− mice. Blood glucose levels were measured at fed (0 h) or fasting conditions (2.5- or 6-h post-fasting). Data were presented as mean ± SD (n = 8, 9, 10, 10, and 10 mice per group for wild-type (WT) treated with PBS, L.G6pc^−/− treated with eGFP, hG6PC S298C mRNA at 0.5 or 1.0 mg/kg, respectively). For statistical analysis, two-sample t-test (two-sided) was performed and corrected for multiple testing by using a Bonferroni adjusted level of 0.005. ^*P ≤ 0.05, ^**P ≤ 0.01, ^***P ≤ 0.001, ^****P ≤ 0.0001, comparing hG6PC S298C mRNA 1.0 mg/kg with eGFP (p values are 0.0017 [day 0, 2.5 h], 0.0016 [day 0, 6 h], 0.0006 [day 2, 2.5 h], 0.0005 [day 2, 6 h], 0.001 [day 4, 2.5 h], 0.001 [day 4, 6 h], and 0.048 [day 7, 6 h], respectively). ^†P ≤ 0.05, ^††P ≤ 0.01, ^†††P ≤ 0.001, ^††††P ≤ 0.0001, comparing hG6PC S298C mRNA 0.5 mg/kg with eGFP (p values are 0.033 [day 0, 2.5 h], 0.001 [day 0, 6 h], 0.0004 [day 2, 2.5 h], 0.00003 [day 2, 6 h], 0.002 [day 4, 2.5 h], 0.001 [day 4, 6 h], and 0.009 [day 7, 6 h], respectively) b Blood glucose levels following repeat (five doses) i.v. administrations of hG6PC S298C mRNA-LNP (0.25 mg/kg) in L.G6pc^−/− mice. Arrows indicate dose administration. Blood glucose levels were measured at 2.5-h post-fasting. Data were presented as mean ± SD (n = 8, 7, and 9 mice per group for WT treated with PBS, L.G6pc^−/− treated with eGFP, and L.G6pc^−/− treated with hG6PC S298C mRNA, respectively). For statistical analysis, two-sample t-test (two-sided) was performed and corrected for multiple testing by using a Bonferroni adjusted level of 0.005. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ***P ≤ 0.0001 comparing hG6PC S298C mRNA with eGFP (p values are 0.005 [day 11], 4 × 10⁻⁶ [day 25], 0.0007 [day 28], 0.0192 [day38], 1.3 × 10⁻⁵ [day 39], 0.001 [day 42], 2 × 10⁻⁶ [day 52], and 5 × 10⁻⁵ [day 53], respectively). c Serum proinflammatory cytokines (from left to right): IFNɣ, IL-1β, TNFα, and IL6 from the dose-ranging study. d serum ALT (mU/mL) levels from the dose-ranging study. For c and d, data were presented as mean ± SD (n = 6, 5, 8, 7, and 6 mice per group for WT treated with PBS, L.G6pc^−/− treated with eGFP, and L.G6pc^−/− treated with hG6PC S298C mRNA at 0.2, 0.5, or 1.0 mg/kg, respectively). e Serum proinflammatory cytokines (from left to right): IFNɣ, IL-1β, TNFα, and IL6 from repeat-dose study. Data were presented as mean ± SD (n = 10, 10, and 7 mice per group for WT treated with PBS, L.G6pc^−/− treated with eGFP, or hG6PC S298C mRNA). f Antidrug antibody assay measuring anti-G6Pase-α antibodies in sera of mice treated with five doses of hG6PC S298C mRNA-LNP (0.5 mg/kg). Data were presented as mean ± SD (n = 9, 7, 7, 6 mice per group for WT treated with PBS, L.G6pc^−/− treated with eGFP, L.G6pc^−/− treated with hG6PC S298C mRNA, and positive sera, respectively). g Body weight of L.G6pc^−/− mice prior to each repeat i.v. dose treatment of hG6PC mRNA -LNP (0.25 mg/kg) for repeat dose study. Data were presented as mean ± SD (n = 8, 7, and 9 mice per group for WT treated with PBS, L.G6pc^−/− treated with eGFP, and L.G6pc^−/− treated with hG6PC S298C mRNA, respectively). For statistical analysis of c–f, raw values were Log2 transformed and subjected to one-way ANOVA, followed by the Dunnett’s multiple comparisons test, compared to the eGFP mRNA treated group. P values are shown in the graphs (c–f). Source data are provided as a Source Data File.