Fig. 7: AAV-based gene therapy approach with shRNA against hBAG3P209L rescues key features of the disease in vivo.
From: Overexpression of human BAG3P209L in mice causes restrictive cardiomyopathy
a Body weight was found to be increased in CAG-BAG3P209L mice 3 weeks after a single intrajugular injection of AAV/rh10 hBAG3 shRNA at P15, when compared to CAG-BAG3P209L mice treated with AAV/rh10 scr. shRNA. Mean ± SEM.; n = 6 CTRL and 8 CAG-BAG3P209L mice. Two-sided Student’s T-test, scr. shRNA = scrambled shRNA. b Immunoblot analysis and quantification of protein expression revealed a decrease in the amount of hBAG3 (BAG3 upper band) in CAG-BAG3P209L mice treated with AAV/rh10 hBAG3 shRNA in comparison to mice treated with AAV/rh10 scr. shRNA. This correlated inversely with the amount of mCherry expression; for normalization of expression levels β-actin was used. Mean ± SEM. n = 6 hearts per group. Two-sided Student’s T-test. scr. RNA = scrambled RNA. c–e Sections of hearts from 5-week-old CAG-BAG3P209L mice which were treated with either AAV/rh10 BAG3 shRNA or AAV/rh10 scr. shRNA were stained for α-actinin (c), desmin (d), FLNC (e). Scale bars: 25 μm. f HE-staining of hearts from 5-week-old CAG-BAG3P209L mice which were treated with either AAV/rh10 hBAG3 shRNA or AAV/rh10 scr. shRNA Scale bar: 100 μm. c–f The experiments were repeated three times from three independent biological replicates with similar results. g Analysis of cardiac fibrosis by Sirius red and Fast green staining after treatment with either AAV/rh10 BAG3 shRNA or AAV/rh10 scr. shRNA. Shown are sections with 33.20% (scr. shRNA treated) and 9.74% (hBAG3 shRNA treated) fibrotic area. h Quantification of the fibrotic area in CAG-BAG3P209L mice 3 weeks after treatment with either scr. or hBAG3 shRNA. Mean ± SEM. n = 12 CTRL and 14 CAG-BAG3P209L mice. Two-sided Student’s T-test. scr. shRNA = scrambled shRNA.
