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

Galina Shapiro, Andrew W. Wong, Maxim Bez, Fang Yang, Sarah Tam, Lisa Even, Dmitriy Sheyn, Shiran Ben-David, Wafa Tawackoli, Gadi Pelled, Katherine W. Ferrara, Dan Gazit

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

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 50 μg luciferase plasmid DNA and 5 × 105 microbubbles followed by ultrasound treatment at 1.4 MHz, 200 kPa, 100-cycle pulse length, and 540 Hz pulse repetition frequency (PRF) for 2 min 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, n = 5 or 6) than the signals for controls treated with DNA injection alone, DNA and microbubble injection, or DNA injection and ultrasound treatment. Our results indicate that these conditions result in efficient gene delivery and prolonged gene expression (up to 21 days) with no evidence of tissue damage or off-target delivery. We believe that these promising results bear great promise for the development of microbubble-enhanced sonoporation-induced gene therapies.

Original languageEnglish (US)
Pages (from-to)157-164
Number of pages8
JournalJournal of Controlled Release
Volume223
DOIs
StatePublished - Feb 10 2016

Fingerprint

Microbubbles
Genes
DNA
Luciferases
Genetic Therapy
Nucleic Acids
Injections
Plasmids
Therapeutics
Cell Membrane
Gene Expression
Thigh
Transgenes
Acoustics
Ultrasonography
Carcinogenesis
Cardiovascular Diseases
Clinical Trials
Safety
Pressure

Keywords

  • "Gene therapy"
  • "Microbubbles"
  • "Sonoporation"
  • "Ultrasound"

ASJC Scopus subject areas

  • Pharmaceutical Science

Cite this

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

In: Journal of Controlled Release, Vol. 223, 10.02.2016, p. 157-164.

Research output: Contribution to journalArticle

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 2016, 'Multiparameter evaluation of in vivo gene delivery using ultrasound-guided, microbubble-enhanced sonoporation', Journal of Controlled Release, vol. 223, pp. 157-164. https://doi.org/10.1016/j.jconrel.2015.12.001
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. / Multiparameter evaluation of in vivo gene delivery using ultrasound-guided, microbubble-enhanced sonoporation. In: Journal of Controlled Release. 2016 ; Vol. 223. pp. 157-164.
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