title: Impact of polyplex micelles installed with cyclic RGD peptide as ligand on gene delivery to vascular lesions.
abstract: Gene therapy is expected to open a new strategy for the treatment of refractory vascular diseases, so the development of appropriate gene vectors for vascular lesions is needed. To realize this requirement with a non-viral approach, cyclo(RGDfK) peptide (cRGD) was introduced to block copolymer, poly(ethylene glycol)-block-polycation carrying ethylenediamine units (PEG-PAsp(DET)). cRGD recognizes α(v)β(3) and α(v)β(5) integrins, which are abundantly expressed in vascular lesions. cRGD-conjugated PEG-PAsp(DET) (cRGD-PEG-PAsp(DET)) formed polyplex micelles through complexation with plasmid DNA (pDNA) and the cRGD-PEG-PAsp(DET) micelles achieved significantly more efficient gene expression and cellular uptake as compared with PEG-PAsp(DET) micelles in endothelial cells and vascular smooth muscle cells. Intracellular tracking of pDNA showed that cRGD-PEG-PAsp(DET) micelles were internalized via caveolae-mediated endocytosis, which is associated with a pathway avoiding lysosomal degradation and that, PEG-PAsp(DET) micelles were transported to acidic endosomes and lysosomes via clathrin-mediated endocytosis. Further, in vivo evaluation in rat carotid artery with a neointimal lesion revealed that cRGD-PEG-PAsp(DET) micelles realized sustained gene expression, whereas PEG-PAsp(DET) micelles facilitated rapid, but transient gene expression. These findings suggest that introduction of cRGD to polyplex micelles might create novel and useful functions for gene transfer and contribute to the establishment of efficient gene therapy for vascular diseases.
authors.affiliation: Division of Vascular Surgery, Department of Surgery, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kagaya
authors.forename: M
authors.initials: M
authors.lastname: Oba
authors.forename: Y
authors.initials: Y
authors.lastname: Miura
authors.forename: H
authors.initials: H
authors.lastname: Koyama
authors.forename: T
authors.initials: T
authors.lastname: Ishii
authors.forename: T
authors.initials: T
authors.lastname: Shimada
authors.forename: T
authors.initials: T
authors.lastname: Takato
authors.forename: K
authors.initials: K
authors.lastname: Kataoka
authors.forename: T
authors.initials: T
authors.lastname: Miyata
mesh_terms: D000818:Animals; D002339:Carotid Arteries; D020212:Carotid Artery Injuries / Q000378:metabolism / Q000628:therapy*; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D008024:Ligands; D008823:Micelles; D023421:Models, Animal; D010456:Peptides, Cyclic / Q000008:administration & dosage* / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011092:Polyethylene Glycols / Q000008:administration & dosage / Q000378:metabolism*; D011506:Proteins / Q000008:administration & dosage / Q000378:metabolism*; D051381:Rats; D019030:Receptors, Vitronectin / Q000378:metabolism
chemical_list: D039302:Integrin alphaVbeta3; D008024:Ligands; D008823:Micelles; D010456:Peptides, Cyclic; D011506:Proteins; D019030:Receptors, Vitronectin; C507169:cyclic arginine-glycine-aspartic acid peptide; C064660:integrin alphaVbeta5; C571441:poly(ethylene glycol)-poly(N'-(N-(2-aminoethyl)-2-aminoethyl)aspartamide) block copolymer; D011092:Polyethylene Glycols