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Optimization of ultrasound parameters for cardiac gene delivery of adenoviral or plasmid deoxyribonucleic acid by Ultrasound-Targeted microbubble destruction

Shuyuan Chen, MD^*, Ralph V. Shohet, MD, FACC^*, Raffi Bekeredjian, MD^*, Peter Frenkel, MD, FACC^*, and Paul A. Grayburn, MD, FACC^{*, ,*}

^* Division of Cardiology, Department of Internal Medicine, University of Texas Southwestern Medical School, Dallas, Texas, USA
Department of Veterans Affairs Medical Center, Dallas, Texas, USA
Baylor University Medical Center, Dallas, Texas, USA

View larger version (17K): [in a new window]   Figure 1 Plot showing the amount of plasmid deoxyribonucleic acid (pDNA) that adheres to washed microbubbles (y-axis) as a function of the amount of pDNA in the solution used to make the microbubbles. Roughly 15% of the DNA adheres to the microbubbles. The total amount of DNA attached to the microbubbles appears to plateau at 4,000 µg/ml of solution.

View larger version (29K): [in a new window]   Figure 2 Plot showing luciferase activity in skeletal muscle, liver, anterior myocardium, and posterior myocardium four days after delivery of microbubbles containing adenovirus encoding the luciferase gene under a cytomegalovirus promoter (ADV) using ultrasound-targeted microbubble destruction. Triggered ultrasound is shown in cross-hatched bars; continuous ultrasound in white bars. Control groups of rats receiving ADV without microbubbles (black bars) and microbubbles without ADV (not shown due to no activity) are also included. Error bars show one standard deviation. US = ultrasound.

View larger version (19K): [in a new window]   Figure 3 Plot showing luciferase activity (y-axis) in skeletal muscle, liver, anterior myocardium, and posterior myocardium at different ultrasound transmission frequencies of 1.3 MHz (white bars), 5 MHz (cross-hatched bars), and 12 MHz (black bars). Error bars show one standard deviation. LV = left ventricle.

View larger version (16K): [in a new window]   Figure 4 Plot showing luciferase activity (y-axis) in skeletal muscle, liver, anterior myocardium, and posterior myocardium during ultrasound-targeted microbubble destruction using ultraharmonic mode (black bars) and harmonic power Doppler (white bars). Images were ECG triggered using a transmission frequency of 1.3 MHz and mechanical index of 1.6. Error bars show one standard deviation. LV = left ventricle.

View larger version (18K): [in a new window]   Figure 5 Plot showing luciferase activity (y-axis) in skeletal muscle, liver, anterior myocardium, and posterior myocardium after ultrasound-targeted microbubble destruction using plasmid deoxyribonucleic acid (pDNA) at different doses of DNA per microbubble solution. Error bars show one standard deviation.

View larger version (15K): [in a new window]   Figure 6 Plot showing luciferase activity (y-axis) in skeletal muscle, liver, anterior myocardium, and posterior myocardium after ultrasound-targeted microbubble destruction using plasmid deoxyribonucleic acid at mechanical index settings of 1.6 (black bars) and 2.0 (white bars). Error bars show one standard deviation. LV = left ventricle.