A new method of ultrasonic nonviral gene delivery to the adult myocardium
| Autor: | Nassim Fares, Anne-Marie Lompré, Adeline Jacquet, Fabrice Atassi, Stéphane N. Hatem, Nathalie Mougenot, Youakim Saliba |
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| Rok vydání: | 2012 |
| Předmět: |
Cardiac fibrosis
Mutant Biology Pharmacology Gene delivery Transfection Mice Transduction (genetics) medicine Animals Myocyte Myocytes Cardiac Ultrasonics Lung Molecular Biology business.industry Myocardium Ultrasound Gene Transfer Techniques beta-Galactosidase medicine.disease Myocardial Contraction Molecular biology Rats medicine.anatomical_structure Liver Ventricle Liposomes Cardiology and Cardiovascular Medicine business Spleen Plasmids |
| Zdroj: | Journal of Molecular and Cellular Cardiology. 53:801-808 |
| ISSN: | 0022-2828 |
| Popis: | Cardiac gene transfer is a powerful molecular tool to improve our understanding of the role of new proteins and mutants in cardiac pathophysiology. There is a need for a simple efficient myocardial gene delivery technique in order to study the physiological role of proteins in their native environment. Here we tested a new method of myocardial nonviral gene delivery, by using the combination of ultrasound energy (USE), liposomes and high pressure injections to the rat heart. Wistar rats were subjected to intra-myocardial injections of liposomes-DNA or siRNA mix. The heart was exposed after an inter-costal incision, and then injections were conducted between two sets of USE heart exposure. Ultrasound application resulted in much higher transfection efficiency (2% of left ventricle) than the liposomes-DNA alone (0.12% of left ventricle) as shown by the beta-galactosidase staining. The ultrasonic based liposomes-DNA delivery resulted in low inflammatory response, as well as in low cardiac fibrosis as shown by total collagen staining. Quantitative real time polymerase chain reaction (PCR) showed that the ultrasonic delivery resulted in cardiac specific transduction. Moreover, 23,906±2197 and 71,883±4065 calcium tolerant transfected cardiac myocytes were isolated following the delivery of a GFP plasmid or tagged siRNA, respectively. This was sufficient to perform single cell physiological measurements and biochemical experiments on homogenates. We developed an interesting safe method for local gene transfer in the heart using ultrasound and liposomes gene delivery. This method is particularly useful to study the effect of gene transfer on cardiac myocytes maintained in their normal environment in animal models. |
| Databáze: | OpenAIRE |
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