Multiparameter evaluation of in vivo gene delivery using ultrasound-guided, microbubble-enhanced sonoporation


Shapiro G, Wong AW, Bez M, Yang F, Tam S, Even L, Sheyn D, Ben-David S, Tawackoli W, Pelled G, Ferrara KW, Gazit D. Multiparameter evaluation of in vivo gene delivery using ultrasound-guided, microbubble-enhanced sonoporation [Internet]. J Control Release 2016;223:157-164. Copy at


{More than 1800 gene therapy clinical trials worldwide have targeted a wide range of conditions including cancer, cardiovascular diseases, and monogenic diseases. Biological (i.e. viral), chemical, and physical approaches have been developed to deliver nucleic acids into cells. Although viral vectors offer the greatest efficiency, they also raise major safety concerns including carcinogenesis and immunogenicity. The goal of microbubble-mediated sonoporation is to enhance the uptake of drugs and nucleic acids. Insonation of microbubbles is thought to facilitate two mechanisms for enhanced uptake: first, deflection of the cell membrane inducing endocytotic uptake, and second, microbubble jetting inducing the formation of pores in the cell membrane. We hypothesized that ultrasound could be used to guide local microbubble-enhanced sonoporation of plasmid DNA. With the aim of optimizing delivery efficiency, we used nonlinear ultrasound and bioluminescence imaging to optimize the acoustic pressure, microbubble concentration, treatment duration, DNA dosage, and number of treatments required for in vivo Luciferase gene expression in a mouse thigh muscle model. We found that mice injected with 50mug luciferase plasmid DNA and 5x10(5) microbubbles followed by ultrasound treatment at 1.4MHz, 200kPa, 100-cycle pulse length, and 540 Hz pulse repetition frequency (PRF) for 2min exhibited superior transgene expression compared to all other treatment groups. The bioluminescent signal measured for these mice on Day 4 post-treatment was 100-fold higher (p<0.0001


1873-4995 Shapiro, Galina Wong, Andrew W Bez, Maxim Yang, Fang Tam, Sarah Even, Lisa Sheyn, Dmitriy Ben-David, Shiran Tawackoli, Wafa Pelled, Gadi Ferrara, Katherine W Gazit, Dan R01CA199658/CA/NCI NIH HHS/United States R01CA103828/CA/NCI NIH HHS/United States R01 CA134659/CA/NCI NIH HHS/United States R01 CA112356/CA/NCI NIH HHS/United States R01CA112356/CA/NCI NIH HHS/United States T32EB003827/EB/NIBIB NIH HHS/United States Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Netherlands J Control Release. 2016 Feb 10;223:157-164. doi: 10.1016/j.jconrel.2015.12.001. Epub 2015 Dec 10.

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