Gene delivery in renal tubular epithelial cells using recombinant adeno-associated viral vectors.
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 x 10(10)/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 S(3) 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 S(3) 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.
Chen
Sifeng
S
Department of Medicine, University of Florida, Gainesville, Florida 32610, USA.
Agarwal
Anupam
A
Glushakova
Olena Y
OY
Jorgensen
Marda S
MS
Salgar
Shashikumar K
SK
Poirier
Amy
A
Flotte
Terence R
TR
Croker
Byron P
BP
Madsen
Kirsten M
KM
Atkinson
Mark A
MA
Hauswirth
William W
WW
Berns
Kenneth I
KI
Tisher
C Craig
CC
D000818:Animals; D000229:Dependovirus / Q000235:genetics*; D004847:Epithelial Cells / Q000502:physiology*; D015870:Gene Expression / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000008:administration & dosage; D007261:Infusions, Intra-Arterial; D007668:Kidney / Q000502:physiology; D007684:Kidney Tubules / Q000502:physiology*; D008164:Luminescent Proteins / Q000008:administration & dosage; D008297:Male; D051381:Rats; D011917:Rats, Inbred Lew; D012077:Renal Artery; D019076:Transgenes / Q000235:genetics*
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins