Focused ultrasound (HIFU) induces localized enhancement of reporter gene expression in rabbit carotid artery.
The development of accurate, safe, and efficient gene delivery remains a major challenge towards the realization of gene therapeutic prevention and treatment of cardiovascular diseases. In this study, we investigated the ability of high-intensity focused ultrasound (HIFU), a form of mechanical wave transmission, to act as a noninvasive tool for the enhancement of in vivo gene transfer into rabbit carotid arteries. Segments of the common carotid arteries of New Zealand white rabbits were isolated and infused with plasmid DNA encoding the reporter beta-galactosidase either with or without the addition of ultrasound contrast agent consisting of small (approximately 2-5 microm) gas-filled human albumin microspheres to augment cavitation. Infused arteries were exposed to pulsed ultrasound for 1 min (frequency 0.85 MHz, burst length 50 ms, repetition frequency 1 Hz, duration 60 s, peak pressure amplitude of 15 MPa). At 6.3 MPa, HIFU enhanced gene expression eight-fold, and 17.5-fold in the presence of contrast. We found increasing amounts of beta-galactosidase expression in the carotid vessel with increasing pressure amplitude. This dose-response relation was present with and without contrast. Without contrast, no vessel damage was detected up to 15 MPa, while the addition of contrast induced side effects above a threshold of 6.3 MPa peak pressure. The entire procedure was feasible and safe for the animals, and the results suggest that HIFU has the potential to assist in the noninvasive spatial regulation of gene transfer into the vascular system.
Huber
P E
PE
Department of Radiology, Brigham and Womens' Hospital, Harvard Medical School, Boston, MA, USA.
Mann
M J
MJ
Melo
L G
LG
Ehsan
A
A
Kong
D
D
Zhang
L
L
Rezvani
M
M
Peschke
P
P
Jolesz
F
F
Dzau
V J
VJ
Hynynen
K
K
D000818:Animals; D002339:Carotid Arteries / Q000201:enzymology*; D002340:Carotid Artery Diseases / Q000628:therapy*; D004247:DNA / Q000008:administration & dosage*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D066298:In Vitro Techniques; D007763:Lac Operon; D008863:Microspheres; D023421:Models, Animal; D011817:Rabbits; D014162:Transfection / Q000379:methods*; D014464:Ultrasonic Therapy / Q000379:methods*
D004247:DNA