Extended Data Fig. 7: Longitudinal assessment of late-stage administration of AAV PKP2 reveals continued effects on alleviating cardiac desmosomal deficits in addition to reduction in cardiac histopathology (fibrosis), arrhythmias and dysfunction at five weeks post-AAV administration.
(a) Schemata for late injection of AAV GFP or AAV PKP2 to PKP2 Hom mice at 4 weeks and post-analysis at 9 weeks (top). Western blot analysis of PKP2 and desmosomal proteins (DSP, DSG2 and JUP) in mouse hearts. β-actin used as a loading control. Data are presented as mean ± S.E.M. n = 6 biologically independent animals per group, Two-way ANOVA with Tukey’s multiple comparison test. *, p < 0.05 (PKP2, p = 0.0410), **, p < 0.01 (DSG2, p = 0.0099; JUP, p = 0.0020). (b) Representative Masson’s Trichrome stains from left (LV) and right ventricular (RV) sections from 9-week-old Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and PKP2 Hom-AAV PKP2 (five weeks post-late stage administration) mice. Scale bar, 2 mm. Experiments were repeated independently three times with similar results. (c) Quantification of mice demonstrating PVCs and sudden death at 9 weeks of age in Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and PKP2 Hom-AAV PKP2 (five weeks post-late stage administration) mice. (d) Quantification of end-diastolic volumes (EDV), end-systolic volumes (ESV) and percentage of ejection fraction (EF %) in 9 week old Ctrl, PKP2 Hom-AAV GFP (five weeks post-late stage administration) and Hom-AAV PKP2 (five weeks post-late stage administration) mice using cardiac MRI. Data are presented as mean ± S.E.M, n = 4 for Ctrl, n = 5 for Hom-AAV GFP group and n = 6 for Hom-AAV PKP2 biologically independent animals. Two-way ANOVA with Tukey’s multiple comparison test. ****, adjusted p < 0.0001. ***, p < 0.001 (EF, LV, p = 0.0008; RV, p = 0.0008). *, p < 0.05 (EDV, LV, p = 0.0369; RV, p = 0.0148; ESV, LV, p = 0.0352), ns, not significant.
