ABSTRACT
Gene therapy has a distinct potential to treat kidney diseases. However, the efficient transduction of a significant number of renal cells by viral vectors has been difficult to accomplish. Previous studies indicate that adeno-associated virus (AAV) can transduce renal cells with variable and suboptimal efficiency. Because new and innovative mutants of AAV are now available, we compared their efficacy in transducing rat kidneys. We compared five types of AAV mutants (AAV2 mut-triple, AAV2 sextuple, AAV8 mut447, AAV8 mut733 and AAV9 mut446) carrying a green fluorescence protein (GFP) reporter gene. A pressure microinjection technique was used to inject either 1.5 × 10(11) vector genome (vg) AAV mutants or three dose of AAV2 sextuple into the renal cortex of rats. The microinjection approach has not been used in AAV-mediated renal gene transfer thus far. Slow and sustained microinjection enables continuous administration of the viral vector to the kidney cortex and limits any damage to the kidney, because the tip of a glass micropipette is very small. Three weeks after injection, the kidneys were collected and evaluated for GFP expression. Among the various mutated AAV serotypes studied, only AAV2 sextuple showed robust GFP expression in renal tissue. The AAV2 sextuple serotype appears to be an efficient gene transfer vector to preferentially target renal tubular epithelial cells. A combination of the AAV2 sextuple and the microinjection technique holds the key to the future of therapeutic treatments for kidney diseases.
