Fig. 4: Nebulized P76 delivery is minimally toxic.
From: Species-agnostic polymeric formulations for inhalable messenger RNA delivery to the lung
aNLuc mRNA was delivered via P76 at 1.25 mg kg–1. Acetate buffer was delivered to the control group. Mice were sacrificed at days 1, 7, 14 and 21 for terminal blood and tissue collection. For each time point, n = 6 mice weights were measured. At the indicated days, blood draws were performed on all the mice; n = 3 mice lungs were used for NanoString analysis and n = 3 mice lungs were used for histopathology analysis. a, Weights as a percentage of starting weight. The error bands represent ±standard error of the mean (s.e.m.). b, Schematic of enzyme-linked immunoassay (ELISA) to detect mouse anti-P76 polyplex antibody responses at days 1, 7, 14 and 21. Horseradish peroxidase (HRP); 3,3′,5,5′-Tetramethylbenzidine (TMB). c, Mouse anti-P76 polyplex antibodies were detected via ELISA. The bars represent mean optical density at 450 nm (OD450). No significant difference was measured by one-way ANOVA (p = 0.6450). d, Complete blood chemistry metrics. *p < 0.05, **p < 0.01 (one-way ANOVA with Dunnett’s multiple comparisons). The grey regions represent 95% confidence interval of naïve mice from Charles River Laboratories. Samples with insufficient sera or excessive haemolysis were excluded from the analysis. The bars in all the graphs indicate mean ± s.d. e, Differential gene expression of 561 inflammatory genes measured by NanoString analysis of the indicated time points versus the control. The horizontal line represents p = 0.05 (two-tailed t-test on log-transformed normalized data) and the vertical lines indicate fold changes of ±2. f, Representative H&E-stained lung sections from mice at the indicated time point. Scale bar, 100 mm. n = 2 mice per group. All the p values are listed in Supplementary Table 3.
