Therapeutic ultrasound optimization for gene delivery: A key factor achieving nuclear DNA localization

Duvshani-Eshet, Maayan; Machluf, Marcelle

doi:10.1016/j.jconrel.2005.08.025

Cited by 93 publications

(80 citation statements)

References 43 publications

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“…The green fluorescence appeared to be in the cytoplasm and excluded from the nucleus. Rapid uptake of naked DNA following sonication has been observed previously [37,38]. When examined after 10 min, sham samples exhibited fluorescence from DNA associated with the plasma membrane (Fig.…”

Section: Effect Of Ultrasound On Intracellular Uptake Of Dna and Pei:supporting

confidence: 61%

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Deshpande

Prausnitz

2007

Journal of Controlled Release

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Ultrasound and poly(ethylenimine) (PEI) have each separately been shown to increase DNA transfection efficiency. This study tested the hypothesis that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection. This in vitro study assessed transfection efficiency of two different DNA plasmids encoding green fluorescent protein and firefly luciferase in two different cells types, a primary culture of human aortic smooth muscle cells and an immortal line of human prostrate cancer cells. We found that ultrasound sonication increased transfection up to 18-fold, DNA complexation with PEI increased transfection up to 90-fold, and the combination of ultrasound and PEI synergistically increased transfection up to 200-fold, which resulted in reporter gene expression by 34% of cells. Kinetic measurements found that the effects of ultrasound alone acted quickly, whereas increased transfection by PEI either alone or in combination with ultrasound strongly benefited from a 4-h incubation with the DNA plasmid after sonication. Although serum reduced absolute expression levels, it did not affect the relative increase in transfection when ultrasound was added to PEI enhancement. Flow cytometry measurements showed that sonication increased intracellular uptake of labelled DNA complexed to PEI by 55% relative to PEI complexation alone. Electrophoresis assay showed no damage to DNA or PEI-DNA complexes after sonication. Overall, these results suggest that the combination of ultrasound and PEI can have a synergistic effect to increase DNA transfection.

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Section: Effect Of Ultrasound On Intracellular Uptake Of Dna and Pei:supporting

confidence: 61%

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Deshpande

Prausnitz

2007

Journal of Controlled Release

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“…7,19 In brief, cells seeded in six-well plates were treated with pDNA coding for luciferase or GFP (7.5 mg/ml) and then exposed to 1 MHz TUS (UltraMax, XLTEK, Canada) at 30% DC, 2 W/cm 2 (correspond to 0.159 MPa or mechanical index (MI) 7 ) for total exposure time of 10, 20 or 30 min. Control cells received only DNA without the application of TUS, or TUS alone.…”

Section: In Vitro Gene Transfectionmentioning

confidence: 99%

“…1 Ultrasound (US) is a relatively new non-viral approach used to deliver genes into cells and tissue. [3][4][5][6][7][8] US waves can be directed to a specific tissue or organ, and can thus localize the DNA of interest and overcome one of the major problems associated with other non-viral vectors for cancer gene therapy. 9,10 Several US modalities are used for gene therapy applications.…”

Section: Introductionmentioning

confidence: 99%

“…9,12,13 Recently, a third US modality, therapeutic ultrasound (TUS), which operates at moderate frequencies of 1-3 MHz and intensities of 0.5-3 W/cm 2 , is being utilized for gene therapy. 7,14 This TUS modality is used mainly for physiotherapy and is approved for clinical applications. 15 The mechanism attributed for US-mediated gene delivery is not fully clear, although cavitation (the creation and oscillation of gas bubbles in a liquid) is suggested to play a major role in cell membrane permeabilization.…”

Section: Introductionmentioning

confidence: 99%

“…Previously, we have shown that long-term TUS (20-30 min) resulted in high transfection in vitro. 7,19 Most importantly, the long-term TUS application was found to enhance DNA trafficking to the nucleus, accelerating its expression to protein. Yet, no study has evaluated the effect and contribution of long-term TUS application with USCA, on expression level and efficiency in vivo in general, and in tumors in particular.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Efficient transfection of tumors facilitated by long-term therapeutic ultrasound in combination with contrast agent: from in vitro to in vivo setting

Duvshani-Eshet

Machluf

2007

Cancer Gene Ther

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Therapeutic ultrasound (TUS) is a promising non-viral clinical approach for the delivery of genes. This study demonstrates the efficient delivery and localization of DNA in subcutaneous tumors facilitated by TUS application and examines the contribution of ultrasound contrast-agent (USCA) addition on transfection. The study addresses the importance of in vivo optimization when using long-term TUS and USCA based on data achieved in vitro. In vitro results showed that transfection of TrampC2 prostate cancer (Pca) cells using genes encoding for luciferase and green fluorescent protein was enhanced when DNA and Optison were added together and TUS was applied for 20 or 30 min. In vivo results showed that the highest transfection was achieved when Optison and DNA were co-injected intratumoraly, and TUS was applied for 20 min. Using Optison significantly increased protein distribution in the tumor. However, in vivo expression level was decreased by two and four fold at 7 and 14 days, respectively, post-TUS. The study establishes the potential of intratumoral delivery of DNA-Optison, followed by TUS as an effective, non-toxic, gene delivery method that could provide a safe, clinical alternative to current viral gene delivery approaches where short-term gene expression is needed.

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Safety and Therapeutic Applications

Azhari¹

2010

Basics of Biomedical Ultrasound for Engineers

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Therapeutic ultrasound optimization for gene delivery: A key factor achieving nuclear DNA localization

Cited by 93 publications

References 43 publications

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Synergistic effect of ultrasound and PEI on DNA transfection in vitro

Efficient transfection of tumors facilitated by long-term therapeutic ultrasound in combination with contrast agent: from in vitro to in vivo setting

Safety and Therapeutic Applications

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