Targeted retroviral gene delivery using ultrasound

Taylor, Stephen K.; Rahim, Ahad A.; Bush, Nigel L.; Bamber, Jeffrey C.; Porter, Colin D.

doi:10.1002/jgm.1003

Cited by 42 publications

(36 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The technique of tuning the ultrasound application to selectively excite microbubbles to control sonoporation should be applicable as a general delivery strategy for many impermeant agents of relevance (12)(13)(14)(15) into selected individual cells with subcellular precision. This approach has the advantage that it requires no physical isolation of cells and/or specialized micronscale fabrication and instrumentation, which are time-consuming and technically challenging.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Spatiotemporally controlled single cell sonoporation

Fan

Liu

Mayer

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

216

215

View full text Add to dashboard Cite

This paper presents unique approaches to enable control and quantification of ultrasound-mediated cell membrane disruption, or sonoporation, at the single-cell level. Ultrasound excitation of microbubbles that were targeted to the plasma membrane of HEK-293 cells generated spatially and temporally controlled membrane disruption with high repeatability. Using whole-cell patch clamp recording combined with fluorescence microscopy, we obtained time-resolved measurements of single-cell sonoporation and quantified the size and resealing rate of pores. We measured the intracellular diffusion coefficient of cytoplasmic RNA/DNA from sonoporation-induced transport of an intercalating fluorescent dye into and within single cells. We achieved spatiotemporally controlled delivery with subcellular precision and calcium signaling in targeted cells by selective excitation of microbubbles. Finally, we utilized sonoporation to deliver calcein, a membrane-impermeant substrate of multidrug resistance protein-1 (MRP1), into HEK-MRP1 cells, which overexpress MRP1, and monitored the calcein efflux by MRP1. This approach made it possible to measure the efflux rate in individual cells and to compare it directly to the efflux rate in parental control cells that do not express MRP1.D espite the development of various approaches for transporting membrane-impermeant compounds (such as fluorescent markers, DNA, RNA, siRNA, proteins, peptides, and amino acids) into living cells (1-3), efficient intracellular delivery of bioactive agents for biomedical applications with minimal adverse effects remains challenging. In addition, it is desirable yet difficult to achieve local perturbation of intracellular processes, which requires subcellular molecular localization inside the living cell (4, 5).Sonoporation uses ultrasound to induce transient disruption of cell membranes (6-8), thereby enabling transport of membraneimpermeant compounds into the cytoplasm of living cells (6,(9)(10)(11). Without the need to use viral vectors, sonoporation enables the delivery of a wide range of bioactive agents with minimal inflammatory and immunological responses for both in vitro studies and in vivo applications (12-16). In addition, ultrasound application can be targeted to a specific volume of tissue in vivo noninvasively. These unique characteristics make sonoporation a compelling and versatile technology for nonviral drug and gene delivery.Sonoporation is typically performed for bulk treatment of a tissue volume in vivo or a large number of cells in vitro, often facilitated by microbubbles that are either injected in the vasculature or mixed in solution with suspended or attached cells. Ultrasound application induces cavitation of the microbubbles (17), signified by rapid volume expansion/contraction and/or collapse (18). These effects generate localized fluid flow, shear stress, and other mechanical or physical impact capable of affecting cells and structures nearby (7,19,20).However, the detailed processes supporting sonoporationmediated transmembrane and ...

show abstract

Section: Resultsmentioning

confidence: 99%

“…Without the need to use viral vectors, sonoporation enables the delivery of a wide range of bioactive agents with minimal inflammatory and immunological responses for both in vitro studies and in vivo applications (12)(13)(14)(15)(16). In addition, ultrasound application can be targeted to a specific volume of tissue in vivo noninvasively.…”

mentioning

confidence: 99%

Spatiotemporally controlled single cell sonoporation

Fan

Liu

Mayer

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

216

215

View full text Add to dashboard Cite

show abstract

“…[22][23][24][25][26][27] Shohet et al 23 demonstrated a successful transfection of rat myocardium in vivo by UTMD of MBs containing an adenovirus encoding a beta-galactosidase reporter gene. Transfection efficiencies by US with retrovirus were enhanced 6.6-fold, 4.8-fold, 2.3-fold and 3.2-fold in 293T cells, BAECs, RASMCs and L6 cells, respectively.…”

Section: Discussionmentioning

confidence: 99%

Ultrasound-targeted microbubble destruction enhances AAV-mediated gene transfection in human RPE cells in vitro and rat retina in vivo

et al. 2009

View full text Add to dashboard Cite

This study was conducted to investigate the efficacy and safety of ultrasound (US)-targeted microbubble (MB) destruction (UTMD)-mediated rAAV2-CMV-EGFP transfection to cultured human retinal pigment epithelium (RPE) cells in vitro and to the rat retina in vivo. In the in vitro study, cultured human RPE cells were exposed to US under different conditions with or without MBs. Furthermore, the effect of UTMD on rAAV2-CMV-EGFP itself and on cells was evaluated. In the in vivo study, gene transfer was examined by injecting rAAV2-CMV-EGFP into the subretinal space of rats with or without MBs and then exposed to US. We investigated enhanced green fluorescent protein (EGFP) expression in vivo by stereomicroscopy and performed quantitative analysis using Axiovision 3.1 software.Hematoxylin and eosin staining and frozen sections were used to observe tissue damage and location of the EGFP gene expression. In the in vitro study, the transfection efficiency of rAAV2-CMV-EGFP under optimal UTMD was significantly higher than that of the control group (P ¼ 0.000). Furthermore, there was almost no cytotoxicity to the cells and to rAAV2-CMV-EGFP itself. In the in vivo study, UTMD could be used safely to enhance and accelerate the transgene expression of the retina. Fluorescence expression was mainly located in the retinal layer. UTMD is a promising method for gene delivery to the retina.

show abstract

“…[29][30][31] Briefly, signal from a waveform generator (model 33220A, Agilent Technologies Inc., Loveland, CO, USA) was fed into an RF power amplifier (58 dB gain, model BT00500 BETA, Tomco Technologies, Norwood, Australia) and thence to a custom-made 1 MHz ultrasound transducer (Imasonics, Besancon, France). The transducer was spherically focused, of 20 mm diameter and 68% bandwidth, with the focal point of the ultrasound beam 67 mm from the transducer face.…”

Section: Ultrasonic Exposure Of the Sie Flapmentioning

confidence: 99%

“…SonoVue is an ultrasound contrast agent consisting of phospholipid-encapsulated microbubbles (1-12 mm) filled with SF 6 gas. 30,31 SonoVue was prepared according to the manufacturer's protocol to yield a solution containing 2 Â 10 8 -5 Â 10 8 microbubbles per ml.…”

Section: Ultrasonic Exposure Of the Sie Flapmentioning

confidence: 99%

Microvascular free tissue transfer for gene delivery: in vivo evaluation of different routes of plasmid and adenoviral delivery

et al. 2008

View full text Add to dashboard Cite

Transfer of healthy autologous tissue as a microvascular free flap facilitates reconstruction during ablative cancer surgery. In addition to filling surgical defects, free flaps might concentrate viral vectors at the tumour bed and mediate local therapeutic effects. We evaluated the magnitude, topography and duration of luciferase gene expression after plasmid and adenoviral delivery in rat superficial inferior epigastric (SIE) flaps. For plasmid delivery, luciferase expression was significantly increased by all transduction routes (topical, intraflap injection, intravascular) ( Po0.01) at day 1, but not at day 7. The spread of luciferase expression was significantly different between the 4 groups at 1 day ( P ¼ 0.026) and was greatest for flaps transduced by intravascular injection. For adenoviral transduction, total radiance was significantly different between the transduced groups at 1, 14 and 28 days (Po0.05 for all comparisons). The highest levels of radiance were seen in the intravascular group. There was a statistically significant difference in the spread of light emission between the 3 groups at 1 ( P ¼ 0.009) and 14 (P ¼ 0.013) days, but this was no longer evident at 28 days. Intravascular adenoviral delivery yields high-level, diffuse and durable gene expression in rat SIE flaps and is suitable for examination in therapeutic models.

show abstract

Targeted retroviral gene delivery using ultrasound

Cited by 42 publications

References 37 publications

Spatiotemporally controlled single cell sonoporation

Spatiotemporally controlled single cell sonoporation

Ultrasound-targeted microbubble destruction enhances AAV-mediated gene transfection in human RPE cells in vitro and rat retina in vivo

Microvascular free tissue transfer for gene delivery: in vivo evaluation of different routes of plasmid and adenoviral delivery

Contact Info

Product

Resources

About