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Targeted Endothelial Gene Delivery by Ultrasonic Destruction of Magnetic Microbubbles Carrying Lentiviral Vectors

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ABSTRACT

Purpose

Site specific vascular gene delivery is a promising tool for treatment of cardiovascular diseases. By combining ultrasound mediated microbubble destruction with site specific magnetic targeting of lentiviruses, we aimed to develop a technique suitable for systemic application.

Methods

The magnetic nanoparticle coupling to lipid microbubbles was confirmed by absorbance measurements. Association of fluorescent lentivirus to magnetic microbubbles (MMB) was determined by microscopy and flow cytometry. Functionality and efficiency of GFP-encoding lentiviral MMB transduction was evaluated by endothelial (HMEC) GFP expression and cytotoxicity was measured by MTT reduction.

Results

Microbubbles with a mean diameter of 4.3 ± 0.04 μm were stable for 2 days, readily magnetizable and magnetically steerable in vitro and efficiently associated with lentivirus. Exposure of eGFP-encoding lentiviral MMB to human endothelial cells followed by application of an external static magnetic field (30 min) and ultrasonic destruction of the microbubbles did not markedly affect cellular viability. Finally, this combination led to a 30-fold increase in transduction efficiency compared to application of naked virus alone.

Conclusions

By associating microbubbles with magnetic iron nanoparticles, these function as carriers for lentiviruses achieving tissue specific deposition at the site of interest.

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Abbreviations

DC:

duty cycle

LV:

lentivirus

MF:

magnetic field

MMB:

magnetic microbubbles

MNP:

magnetic nanoparticles

US:

ultrasound

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ACKNOWLEDGMENTS & DISCLOSURES

This work was funded by the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG) within the DFG Research Unit FOR917.

We thank Professor Kräusslich at the Department of Virology, University Clinic Heidelberg for kindly providing the pCHIV.eGFP lentiviral vector.

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Authors and Affiliations

  1. Walter-Brendel-Centre of Experimental Medicine, Schillerstrasse 44, 80336, Munich, Germany

    Hanna Mannell & Ulrich Pohl

  2. Cardiology, Medical Policlinic, Ludwig-Maximilians-University, Ziemssenstrasse 1, 80336, Munich, Germany

    Joachim Pircher, Thomas Räthel, Katharina Schilberg & Florian Krötz

  3. Institute of Pharmacology and Toxicology, Biomedical Center University of Bonn, Sigmund-Freud-Straße 25, 53105, Bonn, Germany

    Katrin Zimmermann & Alexander Pfeifer

  4. Institute of Experimental Oncology and Therapy Research, Technische Universität München, Ismaninger Str. 22, Munich, 81675, Germany

    Olga Mykhaylyk

  5. Central Institute of Medical Engineering (IMETUM), Technische Universität München, Boltzmannstr. 11, 85748, Garching, Germany

    Bernhard Gleich

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  1. Hanna Mannell
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  2. Joachim Pircher
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Correspondence to Florian Krötz.

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Mannell, H., Pircher, J., Räthel, T. et al. Targeted Endothelial Gene Delivery by Ultrasonic Destruction of Magnetic Microbubbles Carrying Lentiviral Vectors. Pharm Res 29, 1282–1294 (2012). https://doi.org/10.1007/s11095-012-0678-8

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  • DOI: https://doi.org/10.1007/s11095-012-0678-8

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