Cardiac vein retroinjections provide an efficient approach for global left ventricular gene transfer with adenovirus and adeno-associated virus

Figures

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  Figure 1. Catheterization setup. (A) Schematic about the catheterization setup of the method. The retroinjection was given through the wedge pressure balloon catheter placed in the principal vein of the target area, during simultaneous occlusion of the supplying coronary artery of the target area. Also, the main anastomotic vein of the target area can be occluded. (B) To treat the LVAW and anterior IVS, retroinjection was given into the ACV, during simultaneous transient LAD occlusion. (C) To treat the LVLW, retroinjection was given into the most prominent vein of the region (LMV or LPMV), and LCx was occluded transiently during the retroinjection. (D) To treat the LVPW and posterior IVS, retroinjection was given to the MCV. During retroinjection,either RCA (1.) or LCx (2.), depending of which one is dominant in posterior LV, was occluded. (B,C,D) In some animals, the main anastomotic vein of each retroinjected vein was occluded during the retroinjections. (B,C,D) The vessel of retroinjection is marked with a syringe symbol.

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  Figure 2. Sampling sites of the cardiac ventriculi. Of each site, both histological and snap-frozen samples were collected.

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  Figure 3. Beta-galactosidase immunostainings of LVAW and IVS from animals of the first set of experiments. The samples on the left are from the animal that underwent ACV retroinjection without LAD occlusion. On the right are samples from the animal that underwent ACV retroinjection with transient LAD occlusion. The sections are photographed at 20 × magnification. Samples were collected from the LVAW and the IVS as presented in Fig. 2.

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  Figure 4. Ink distribution after different retroinjection procedures. The heart slices are oriented as anterior wall upwards. (A) 10 ml retroinjection into the LMV with a simultaneous LCx occlusion. (B) 10 ml retroinjection into the LMV with simultaneous LCx and MCV occlusions. (C) 10 ml retroinjection into the MCV with a simultaneous RCA occlusion. (D) 10 ml retroinjection into the MCV with simultaneous RCA and ACV occlusions. (E) 20 ml retroinjection into the ACV with a simultaneous LAD occlusion. (F) 20 ml retroinjection into the ACV with simultaneous LAD and MCV occlusions. (G) 20 ml retroinjection into the ACV with simultaneous LAD and MCV occlusions. Half of the ink solution was administered through a microcatheter placed in the apical part of the ACV. (H) Distribution outcome of retroinjections into the three main LV veins, with simultaneous transient occlusions of the coronary arteries and the main anastomotic branches of the cardiac veins. The retroinjections were performed as described in the Fig. 1.

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  Figure 5. Vector genome distribution in heart and extracardiac tissues. Figures A-C are from AAV2-GFP animals at 1 month timepoint. Figures D-F are from AdLacZ animals at six-day timepoint. (A) AAV genome intracardiac distribution. (B) Vertical LV distribution of AAV genome. (C) AAV genome biodistribution. (D) Ad genome intracardiac distribution. (E) Vertical LV distribution of Ad genome. (F) Ad genome biodistribution. LV mean includes and RV mean excludes the IVS. Each dot indicates one animal’s mean value of the collected samples from the area in question. LN indicates lymph node. Cardiac sampling sites are presented in Fig. 2. Closed circles: LV; open circles: RV.

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  Figure 6. Clinical chemistry. Results are presented as mean ± SEM. * indicates statisticalyy significant (p < 0.05) difference between measurements indicated in square brackets. Grey columns indicate the results of AdLacZ animals. White columns indicate the results of AAV2-GFP animals.