Gene Delivery in Renal Tubular Epithelial Cells Using Recombinant Adeno-Associated Viral Vectors

Abstract

ABSTRACT. Gene therapy has the potential to provide a therapeutic strategy for numerous renal diseases such as diabetic nephropathy, chronic rejection, Alport syndrome, polycystic kidney disease, and inherited tubular disorders. In previous studies using cationic liposomes or adenoviral or retroviral vectors to deliver genes into the kidney, transgene expression has been transient and often associated with adverse host immune responses, particularly with the use of adenoviral vectors. The unique properties of recombinant adeno-associated viral (rAAV) vectors permit long-term stable transgene expression with a relatively low host immune response. The purpose of the present study was to evaluate gene expression in the rat kidney after intrarenal arterial infusion of a rAAV (serotype 2) vector encoding green fluorescence protein (GFP) induced by a cytomegalovirus-chicken beta-actin hybrid promoter. The left kidney of experimental animals was treated with either saline or transduced with rAAV2-GFP (0.125 ml/100 g body wt, 1 × 1010/ml infectious units) through the renal artery. A time-dependent expression of GFP was observed in all kidneys injected with rAAV2-GFP, with maximal expression observed at 6 wk posttransduction. The expression of GFP was restricted to cells in the S3 segment of the proximal tubule and intercalated cells in the collecting duct, the latter identified by co-localization with H+-ATPase. No transduction was observed in the glomeruli or the intrarenal vasculature. These studies demonstrate successful transgene expression in tubular epithelial cells, specifically in the S3 segment of the proximal tubule and intercalated cells, after intrarenal administration of a rAAV vector and provide the impetus for further studies to exploit its use as a tool for gene therapy in the kidney.E-mail: agarwal@nersp.nerdc.ufl.edu

Perspectives for Gene Therapy in Renal Diseases

“…The expression lasted at least 3 months. Similar results were obtained by Chen et al (24). Gene transduction and expression were timedependent and restricted to cells of S3 segment of the proximal tubule and intercalated cells in the collecting duct.…”

Adeno-Associated Virus–Mediated CTLA4Ig Gene Transfer Protects MHC-Mismatched Renal Allografts from Chronic Rejection

“…Intrarenal arterial administration of rAAV that carried green fluorescence protein DNA in the rat resulted in transduction of renal tubular epithelial cells, with the transgene expressed for 6 wk and a relatively modest host immune response (14). Furthermore, injection of rAAV5 encoding reporter genes into the mouse renal parenchyma yielded transfection of tubular epithelial cells in the vicinity of the injection site as long as 3 mo (15).…”

Transduction of interleukin-10 through renal artery attenuates vascular neointimal proliferation and infiltration of immune cells in rat renal allograft

“…The left kidney of BN donor was transplanted to the WF recipient in accordance with the method described by Lee [20]. Before engrafting, the donor BN kidney allograft received in situ perfusion of 0.5ml cold PBS (n=6), 0.5ml cold PBS with rAAv1-IL-10 (n=5) or rAAv1-YFP (n=5 including one died before sacrificed due to immune rejection) at a dose of 8×109 vial particles/kidney through renal artery according to the method we described previously [21]. All perfusion solutions contained heparin (40 units/ml).…”