Figure 1: rAAV9-BAG3 Improves LV Function in Mice With LV Dysfunction Secondary to Ligation of the Left Coronary Artery
2019-06-11T07:03:08Z (GMT) by
(A) Western blot analysis of tissue obtained on various days from mice after retro-orbital injection of myc−-tagged adeno-associated virus serotype 9 expressing BAG3 (rAAV9-BAG3). (B) Eight-week-old C57BL/6J were subjected to ligation of the left coronary artery (myocardial infarction [MI]) or sham surgery (Sham) (week 0, arrow A). The first 1 week post-surgical echocardiogram is indicated by arrow B. At 8 weeks post-surgery (arrow C), the MI and Sham groups were randomly assigned to receive 60 to 80 μl of either rAAV9-GFP (3.1 × 1012 genome copies [GC]/ml; Sham- or MI-GFP) or rAAV9-BAG3 (5 to 6.5 × 1013 GC/ml; Sham- or MI-BAG3) by injection into the retro-orbital venous plexus. Serial echocardiography was performed to determine the ejection fraction (EF), including 1 week before and 9 days and 3 weeks (arrow D) after injection. Data was analyzed using a 2-way analysis of variance for repeated measures. n = 14 Sham-GFP, n = 12 Sham-BAG3, n = 12 MI-GFP, and n = 13 MI-BAG3. ∗MI-GFP versus -BAG3 at week 9, p < 0.0001; †MI-GFP versus -BAG3 at week 11, p < 0.0001. There was no statistical difference between Sham-GFP and -BAG3 at any time points. (C) Western blot analysis of BAG3 levels in right ventricles of MI-GFP, MI-BAG3, and Sham-GFP mice (n = 3) sacrificed 11 weeks post-MI (3 weeks post-injection of rAAV9-BAG3 or -GFP) ∗p < 0.04; MI-GFP compared with MI-BAG3 or Sham-GFP. Confocal images demonstrating normal BAG3 levels in Sham-GFP hearts (D) reduced BAG3 levels in MI-GFP myocytes (E) and reconstitution and correct targeting of exogenous BAG3 in MI-BAG3 myocytes (F).