title: DNA/dendrimer complexes mediate gene transfer into murine cardiac transplants ex vivo.
abstract: Starburst polyamidoamine dendrimers are synthetic polymers with unique structural and physical characteristics suitable for DNA gene transfer. Our previous studies demonstrated that Starburst dendrimers augment plasmid-mediated gene transfer efficiency in a nonvascularized, cardiac transplantation model. In this study, the fifth generation of ethylenediamine core dendrimer was investigated for its ability to enhance gene transfer and expression in a clinically relevant murine vascularized heart transplantation model. The plasmid pMP6A-beta-gal, encoding beta-galactosidase (beta-Gal), was incubated with dendrimers to form complexes. The complexes were perfused via the coronary arteries during donor graft harvesting, and reporter gene expression was determined by quantitative evaluation of X-Gal staining. The grafts infused with pMP6A-beta-gal/dendrimer complexes showed beta-Gal expression in myocytes from 7 to 14 days. A number of variables for transfer of the DNA/dendrimer complexes were tested, including DNA:dendrimer charge ratios, concentrations of DNA and dendrimer, preservation solutions, ischemic time, and enhancement of vascular permeability by serotonin, papaverine, and VEGF administration. The results showed that DNA/dendrimer complexes containing 20 microg of DNA and 260 microg of dendrimer (1:20 charge ratio) in a total volume of 200 microl resulted in highest gene expression in the grafts. The results also showed that prolonged incubation (cold ischemic time) to 2 h and pretreatment with serotonin further enhanced gene expression.
authors: Institute for Gene Therapy and Molecular Medicine, Mount Sinai School of Medicine, New York, New York 10029, USA.
authors: Y
authors: Y
authors: Wang
authors: P
authors: P
authors: Boros
authors: J
authors: J
authors: Liu
authors: L
authors: L
authors: Qin
authors: Y
authors: Y
authors: Bai
authors: A U
authors: AU
authors: Bielinska
authors: J F
authors: JF
authors: Kukowska-Latallo
authors: J R
authors: JR
authors: Baker
authors: J S
authors: JS
authors: Bromberg
mesh_terms: D000818:Animals; D004247:DNA / Q000235:genetics*; D016228:Endothelial Growth Factors / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D018014:Gene Transfer Techniques; D016027:Heart Transplantation; D066298:In Vitro Techniques; D008222:Lymphokines / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D010208:Papaverine / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D012701:Serotonin / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D016228:Endothelial Growth Factors; D008222:Lymphokines; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D012701:Serotonin; D011092:Polyethylene Glycols; D004247:DNA; D010208:Papaverine; D001616:beta-Galactosidase; C016811:tyloxapol