Efficient in vivo gene transfer to murine renal cells using AAV9 vectors (912.7)

Adeno‐associated viral (AAV) vectors provide sustained long‐term gene expression with minimal immunological consequences. Therefore AAV vector systems are useful gene‐transfer tools in translational, as well as in basic research studies. Systemic administration of AAV9 vector transduces various tissues efficiently but provides very low transduction in kidney, compared to other organs such as liver, heart, and skeletal muscle. We hypothesized that the administration of AAV9 vector by retrograde infusion via the ureter can provide efficient gene transfer to renal cells and minimize off‐target gene expression. AAV9 vector (1e+11 viral genome particles/100 μl), encoding firefly luciferase (AAVCMVLuc) or eGFP (AAVCMVeGFP) under the control of CMV promoter, was infused into the left kidney via the ureter and held for a period of 15 minutes. Non‐invasive bioluminescence imaging (IVIS‐200, Xenogen Corporation) of the mice treated with AAVCMVLuc after one week showed that the signal was limited to the left kidney. Three weeks following vector administration, the magnitude and distribution of gene expression was measured by luciferase activity assay and immunohistochemistry, respectively. Luciferase activity was 4.6‐fold (n=3, P<0.05) higher in the kidney infused with AAVCMVLuc than the liver. Immunohistochemical analyses of AAVCMVeGFP‐treated mice showed gene expression in renal distal, as well as proximal tubular epithelial cells. These studies show that AAV9 gene transfer via the ureter is a highly efficient method for gene transfer and gene rescue studies in mouse kidney. Grant Funding Source : 1R01DK090918‐01A1