FIG. 7.

Proteasome inhibitors augment rAAV transduction to mouse lungs and human bronchial xenografts in vivo. (A) Aerosol delivery of rAAV with Z-LLL or doxorubicin to the mouse airway. C57BL/6 mice were infected with a total of 6 × 10¹⁰ particles of AV2.Luc or AV2/5.Luc through nasal aspiration in the absence or presence of either 400 μM Z-LLL or 200 μM doxorubicin (DOX). Viruses and the proteasome inhibitor were mixed together (40 μl of inoculum/mouse) and delivered through the nose three times on sequential days. Two weeks after the final infection, the mouse lungs and trachea were harvested and homogenized in a Promega luciferase reporter lysis buffer. Samples were normalized for protein concentration, and luciferase activity was measured in a luminometer at 80% sensitivity. The values represent the mean (± standard error of the mean) relative luciferase activities in both the lungs and trachea (n = 3). (B) Human bronchial xenografts were infected with 10¹¹ particles (in a volume of 100 μl) of AV2.Luc. The administration of proteasome inhibitors was either through a local application mixed with virus and applied to the lumen of grafts at the time of infection (doxorubicin and Z-LLL) or through systemic application to the mouse by tail vein injection 1 and 2 days after rAAV infection of the airway lumen (Doxil). Total microgram doses and working concentrations of drugs are summarized below the chart (+, present; −, absent). The xenograft airways were harvested at 2 weeks postinfection, and luciferase assays were performed on the entire airway. Results depict the means ± standard errors of the means for three independent xenografts for each experimental point.