Gene transfer with synthetic virus-like particles via the integrin-mediated endocytosis pathway.
The interaction between cationic DNA-containing particles and cell surface anionic proteoglycans is an efficient means of entering cultured cells. Therapeutic in vivo gene delivery levels, however, require binding to less ubiquitous molecules. In an effort to follow adenovirus, thiol-derivatized polyethylenimine (PEI) was conjugated to the integrin-binding peptide CYGGRGDTP via a disulfide bridge. The most extensively conjugated derivative (5.5% of the PEI amine functions) showed physical properties of interest for systemic gene delivery. In the presence of excess PEI-RGD, plasmid DNA was condensed into a rather homogeneous population of 30-100 nm toroidal particles as revealed by electron microscopy images in 150 mM salt. Their surface charge was close to neutrality as a consequence of the shielding effect of the prominent zwitterionic peptide residues. Transfection efficiency of integrin-expressing epithelial (HeLa) and fibroblast (MRC5) cells was increased by 10- to 100-fold as compared with PEI, even in serum. This large enhancement factor was lost when aspartic acid was replaced by glutamic acid in the targeted peptide sequence (RGD/RGE), confirming the involvement of integrins in transfection. PEI-RGD/DNA complexes thus share with adenovirus constitutive properties such as size and a centrally protected DNA core, and 'early' properties, i.e. cell entry mediated by integrins and acid-triggered endosome escape.
Erbacher
P
P
Laboratoire de Chimie Génétique, UMR 7514 CNRS/Université Louis Pasteur de Strasbourg, Faculté de Pharmacie, Illkirch, France.
Remy
J S
JS
Behr
J P
JP
D002412:Cations; D005347:Fibroblasts; D018014:Gene Transfer Techniques; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D016023:Integrins / Q000235:genetics*; D008156:Luciferases / Q000235:genetics; D008854:Microscopy, Electron; D011094:Polyethyleneimine
D002412:Cations; D016023:Integrins; D011094:Polyethyleneimine; D008156:Luciferases