Fig. 3: AAV.FUS candidates improve efficiency of gene delivery to the brain and reduce peripheral transduction.
From: Engineering viral vectors for acoustically targeted gene delivery
a Representative images were obtained from mice co-injected with AAV9 and a AAV.FUS.3 at 1010 viral genomes per gram of body weight each. Sections show brain transduction by AAV9 (red) and AAV.FUS.3 (green), and are counterstained with a neuronal stain (NeuN, blue). b All but one (AAV.FUS.4) AAV.FUS candidates showed significant improvement over the co-injected AAV9. (p-values for AAV.FUS.1-5, p = 0.0274, 0.0003, 0.0052, 0.2556, 0.0087, respectively; Two-way ANOVA with Sidak multiple comparisons test: F(1, 24) = 59.49, P value; P < 0.0001). Data from 3 male and 3 female mice per serotype. c We found that few cells were transduced outside of the FUS-targeted site and AAV.FUS.3 and AAV9 were not significantly different. (0.19% vs 0.4%, respectively; p = 0.072, two-way ANOVA with Sidak multiple comparisons test; F(1, 35) = 2.457, p = 0.1260). Similarly, other candidates also showed no differences compared to AAV9 (AAV.FUS.1, p = 0.99; AAV.FUS.2, p = 0.98; AAV.FUS.4, p = 0.86; AAV.FUS.5, p = 0.83). Data from 3 male and 3 female mice for all serotypes, except AAV.FUS.2 (2 male, 3 female mice), and AAV.FUS.3 (8 male and 8 female mice). d Representative images showing liver transduction by AAV9 (red) and AAV.FUS.3 (green). e All tested candidates showed reduced liver transduction as compared to the co-injected AAV9 in the same mice for which brain expression was analyzed. (P-values for AAV9 vs AAV.FUS.1-5 were p = 0.0058 for AAV.FUS.1, and p < 0.0001 for other candidates; Two-way ANOVA, F(1, 24) = 375.9, P < 0.0001. Data from 3 male and 3 female mice for all serotypes, except AAV.FUS.2 (2 male, 3 female mice). f We defined the fold-improvement in targeting efficiency as the ratio of brain transduction to the liver transduction efficiency using AAV9 as a baseline, which suggested that AAV.FUS.3 is the top candidate for further study. (AAV.FUS.3 compared to AAV.FUS.1,2,4,5, all p-values were p < 0.0001, one way ANOVA with Tukey HSD post hoc comparison test). Scale bars are 50 μm in (a, c), unless otherwise noted. (****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05, ns = not significant); Error bars are 95% CI. The numbers of animals used in each experiment were: Data from 3 male and 3 female mice per serotype, except AAV.FUS.2 (2 male, 3 female mice). Center for the error bars represents arithmetic mean in (b, c, e, f).
