Glomerular filtration is required for transfection of proximal tubular cells in the rat kidney following injection of DNA complexes into the renal artery.
Gene transfer to the kidney can be achieved with various DNA vectors, resulting in transgene expression in glomerular or tubular districts. Controlling transgene destination is desirable for targeting defined renal cells for specific therapeutic purposes. We previously showed that injection of polyplexes into the rat renal artery resulted in transfection of proximal tubular cells. To investigate whether this process involves glomerular filtration of the DNA-containing particles, fluorescent polyethylenimine polyplexes were prepared, containing fluoresceinated poly-L-lysine. This allowed visualization of the route of the particles into the kidney. Our polyplexes were filtered through the glomerulus, since fluorescent proximal tubuli were observed. Conversely, fluorescent lipopolyplexes containing the cationic lipid DOTAP were never observed in tubular cells. Size measurements by laser light scattering showed that the mean diameter of polyplexes (93 nm) was smaller than that of lipopolyplexes (160 nm). The size of the transfecting particles is therefore a key parameter in this process, as expected by the constraints imposed by the glomerular filtration barrier. This information is relevant, in view of modulating the physico-chemical properties of DNA complexes for optimal transgene expression in tubular cells. Gene Therapy (2000) 7, 279-285.
Foglieni
C
C
Biotechnology Unit, Dibit, Department of Biological and Technological Research, San Raffaele Scientific Institute, Milan, Italy.
Bragonzi
A
A
Cortese
M
M
Cantù
L
L
Boletta
A
A
Chiossone
I
I
Soria
M R
MR
Monaco
L
L
D000818:Animals; D004247:DNA / Q000235:genetics*; D005229:Fatty Acids, Monounsaturated / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D005822:Genetic Vectors / Q000235:genetics*; D007678:Kidney Glomerulus / Q000502:physiology*; D007687:Kidney Tubules, Proximal / Q000502:physiology*; D011107:Polylysine / Q000235:genetics / Q000493:pharmacokinetics; D000644:Quaternary Ammonium Compounds / Q000493:pharmacokinetics; D051381:Rats; D014162:Transfection / Q000235:genetics*; D001616:beta-Galactosidase / Q000378:metabolism
D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D000644:Quaternary Ammonium Compounds; D011107:Polylysine; D004247:DNA; D001616:beta-Galactosidase; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane