Ultrasound-Mediated Vascular Gene Transfection by Cavitation of Endothelial-Targeted Cationic Microbubbles

JACC-CARDIOVASCULAR IMAGING

卷:5

Article

Xie, Aris¹  Belcik, Todd¹  Qi, Yue¹  Morgan, Terry K.²  Champaneri, Shivam A.³  Taylor, Sarah³  Davidson, Brian P.¹  Zhao, Yan¹  Klibanov, Alexander L.³  Kuliszewski, Michael A.⁴  Leong-Poi, Howard⁴  Ammi, Azzdine¹  Lindner, Jonathan R.¹ 

[1] Oregon Hlth & Sci Univ, Div Cardiovasc Med, Portland, OR 97239 USA

[2] Oregon Hlth & Sci Univ, Dept Pathol, Portland, OR 97239 USA

[3] Univ Virginia, Div Cardiovasc, Charlottesville, VA USA

[4] St Michaels Hosp, Toronto, ON M5B 1W8, Canada

关键词: contrast ultrasound;    gene delivery;    microbubbles;   

DOI  :  10.1016/j.jcmg.2012.05.017

来源: Elsevier

【 摘 要 】

OBJECTIVES Ultrasound-mediated gene delivery can be amplified by acoustic disruption of microbubble carriers that undergo cavitation. We hypothesized that endothelial targeting of microbubbles bearing cDNA is feasible and, through optimizing proximity to the vessel wall, increases the efficacy of gene transfection. BACKGROUND Contrast ultrasound-mediated gene delivery is a promising approach for site-specific gene therapy, although there are concerns with the reproducibility of this technique and the safety when using high-power ultrasound. METHODS Cationic lipid-shelled decafluorobutane microbubbles bearing a targeting moiety were prepared and compared with nontargeted microbubbles. Microbubble targeting efficiency to endothelial adhesion molecules (P-selectin or intercellular adhesion molecule [ICAM]-1) was tested using in vitro flow chamber studies, intravital microscopy of tumor necrosis factor-alpha (TNF-alpha)-stimulated murine cremaster muscle, and targeted contrast ultrasound imaging of P-selectin in a model of murine limb ischemia. Ultrasound-mediated transfection of luciferase reporter plasmid charge coupled to microbubbles in the post-ischemic hindlimb muscle was assessed by in vivo optical imaging. RESULTS Charge coupling of cDNA to the microbubble surface was not influenced by the presence of targeting ligand, and did not alter the cavitation properties of cationic microbubbles. In flow chamber studies, surface conjugation of cDNA did not affect attachment of targeted microbubbles at microvascular shear stresses (0.6 and 1.5 dyne/cm(2)). Attachment in vivo was also not affected by cDNA according to intravital microscopy observations of venular adhesion of ICAM-1-targeted microbubbles and by ultrasound molecular imaging of P-selectin-targeted microbubbles in the post-ischemic hindlimb in mice. Transfection at the site of high acoustic pressures (1.0 and 1.8 MPa) was similar for control and P-selectin-targeted microbubbles but was associated with vascular rupture and hemorrhage. At 0.6 MPa, there were no adverse bioeffects, and transfection was 5-fold greater with P-selectin-targeted microbubbles. CONCLUSIONS We conclude that ultrasound-mediated transfection at safe acoustic pressures can be markedly augmented by endothelial juxtaposition. (J Am Coll Cardiol Img 2012; 5: 1253-62) (C) 2012 by the American College of Cardiology Foundation

【 授权许可】

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