DNA/dendrimer complexes mediate gene transfer into murine cardiac transplants ex vivo.

TitleDNA/dendrimer complexes mediate gene transfer into murine cardiac transplants ex vivo.
Publication TypeJournal Article
Year of Publication2000
AuthorsWang Y, Boros P, Liu J, Qin L, Bai Y, Bielinska AU, Kukowska-Latallo JF, Baker JR, Bromberg JS
JournalMol Ther
Date Published2000 Dec
KeywordsAnimals, beta-Galactosidase, DNA, Endothelial Growth Factors, Gene Expression Regulation, Gene Transfer Techniques, Heart Transplantation, In Vitro Techniques, Lymphokines, Mice, Mice, Inbred BALB C, Papaverine, Polyethylene Glycols, Serotonin, Vascular Endothelial Growth Factor A, Vascular Endothelial Growth Factors

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.

Alternate JournalMol Ther
PubMed ID11124061
Grant ListAI42840 / AI / NIAID NIH HHS / United States
Related Faculty: 
Lihui Qin, M.D., Ph.D.

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