Efficient transfer of oligonucleotides and plasmid DNA into the whole heart through the coronary artery

Abstract

Several of the current techniques for transfer of both olidetected in the nuclei of more than 70% of the myocytes gonucleotide and plasmid DNA into the myocardium are and endothelial cells both in the epicardium and endocarimpaired by low efficiency and toxicity. To improve gene dium. ␤-Gal was expressed in the cytosol of more than transfer techniques, especially into the whole heart, a gene 50% of the myocytes. ␤-Gal expression was demonstrated transfer method involving liposome in conjunction with a by Western blotting analysis at day 3 after transfection and viral envelope (HVJ-liposome) was essayed as an alternacontinued for at least 14 days. No significant histological tive. FITC-labeled oligonucleotide (F-ODN) and the cDNA damage of the myocardium or leakage of CPK were of ␤-galactosidase (␤-gal) were introduced into the myocardetected in the rats transfected by the HVJ-liposome dium by coronary infusion of HVJ-liposome during cardiomethod. These results clearly demonstrate that the hearts plegic arrest of adult Sprague-Dawley rat hearts. Then, were efficiently transfected both by oligonucleotide and transfected heart was ectopically transplanted into another plasmid DNA as a result of coronary infusion of HVJ-liporat abdomen of the same strain to maintain the transfected some during cardioplegic arrest. This thus appears to be heart long enough to allow for protein synthesis. After 3 an efficient method for gene transfer into the whole heart, days of transfection, transfected heart was excised and the providing a new tool for research and therapy for heart efficiency of gene transfection was evaluated. FITC was diseases.

Nonviral gene transfer to skeletal, smooth, and cardiac muscle in living animals

“…However, infusion through the coronary artery or aorta in vivo (i.e., an in situ heart) appeared to result in much less efficient uptake of DNA (Ͻ1.6% of cardiomyocytes) (33,109). Unlike the study of Sawa et al (105) using HVJ liposomes in excised, transplanted hearts, the use of other lipids [N,N-dimethyl-2,3-bis(dodecyloxy)-1-propanaminium bromide (DLRIE)-DOPE] or dendrimers to complex with the DNA resulted in very low levels of gene transfer to either endothelial cells or cardiomyocytes (Ͻ1% of cells) when a similar infusion/transplant method was used (24,108,125).…”

“…DNA has been complexed with a number of different lipids for gene transfer to the heart via the circulation. A number of studies have used a mixture of phosphatidylserine, cholesterol, and phosphatidylcholine mixed with inactivated hemagglutinating virus of Japan (HVJ) particles (33,105,112). These HVJ liposomes are fusogenic and promote fusion of the liposome with the endosomal membrane and uptake of the DNA into the cell.…”

“…These HVJ liposomes are fusogenic and promote fusion of the liposome with the endosomal membrane and uptake of the DNA into the cell. In several studies, mixtures of plasmid and HVJ liposomes were infused over the course of 10 min into either the aorta or coronary arteries of excised hearts maintained at 4°C, and then the hearts were transplanted into recipient mice or rats (105,112). In one study, this method resulted in up to 50% of cardiomyocytes receiving and expressing the transgene for up to 14 days (the ␤-actin promoter was used to drive gene expression in these studies) (105).…”

“…In several studies, mixtures of plasmid and HVJ liposomes were infused over the course of 10 min into either the aorta or coronary arteries of excised hearts maintained at 4°C, and then the hearts were transplanted into recipient mice or rats (105,112). In one study, this method resulted in up to 50% of cardiomyocytes receiving and expressing the transgene for up to 14 days (the ␤-actin promoter was used to drive gene expression in these studies) (105). However, infusion through the coronary artery or aorta in vivo (i.e., an in situ heart) appeared to result in much less efficient uptake of DNA (Ͻ1.6% of cardiomyocytes) (33,109).…”

In vivo myocardial gene transfer: optimization and evaluation of intracoronary gene delivery in vivo

“…The potential of intracoronary gene delivery has been demonstrated ex vivo. 6 However, the technique used has limited clinical applicability. Initial results with intracoronary gene transfer in vivo appeared promising, with efficient adenoviral-mediated gene transfer to 30% of myocytes in a vascular territory of the rabbit heart.…”