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Fatty acid–spermine conjugates as DNA carriers for nonviral in vivo gene delivery

Gene Therapy volume 16, pages 1429–1440 (2009)Cite this article

Abstract

The lack of efficient in vivo gene delivery is a well-known shortcoming of nonviral delivery vectors, in particular of chemical vectors. We developed a series of novel nonviral carriers for plasmid-based in vivo gene delivery. This new transport device is based on the assembly of DNA plasmids with synthetic derivatives of naturally occurring molecules—fatty acid–spermine conjugates (or lipospermines). We tested the ability of these fatty acid conjugates to interact with plasmid DNA (pDNA) and found that they formed DNA nanocomplexes, which are protected from DNase I degradation. This protection was shown to directly correlate with the length of the aliphatic component. However, this increase in the length of the hydrocarbon chain resulted in increased toxicity. The cationic lipids used for transfection typically have a C16 and C18 hydrocarbon chain. Interestingly, toxicity studies, together with further characterization studies, suggested that the two most suitable candidates for in vivo delivery are those with the shortest hydrocarbon chain, butanoyl- and decanoylspermine. Morphological characterization of DNA nanocomplexes resulting from these lipospermines showed the formation of a homogenous population, with the diameter ranging approximately from 40 to 200 nm. Butanoylspermine was found to be the most promising carrier from this series, resulting in a significantly increased gene expression, in relation to naked plasmid, in both tissues herein targeted (dermis and M. tibialis anterior). Thus, we established a correlation between the in vitro properties of the ensuing DNA nanocarriers and their efficient in vivo gene expression.

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Acknowledgements

We thank Alex Perálvarez-Marín for his invaluable help in dynamic light scatter measurements. We also thank the Organic Chemistry Department and Professor Astrid Gräslund from the Department of Biochemistry and Byophysics at Stockholm University for providing access to the zetasizer equipment. This project was financially supported by the Swedish Research Council, The Swedish Foundation for Strategic Research Bio-Xgrant, KI Faculty funds for funding of postgraduate students, Marie Curie fellowship programme—Eurogendis training site, Portuguese Foundation for Science and Technology (SFRH-BD-16757-2004), the European Union FP6 grant EuroPharmacoGene (FP6-2005-037283) and Synthe Gene Delivery (LSHB-CT-2005-018716).

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Authors and Affiliations

  1. Department of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, Karolinska University Hospital Huddinge, Stockholm, Sweden

    J R Viola, O E Simonson, I I Oprea, P M D Moreno, K E Lundin & C I E Smith

  2. Department of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden

    H Leijonmarck, P Purhonen & R Strömberg

  3. Department of Physiology and Pharmacology, Karolinska Institutet, Stockholm, Sweden

    R Frithiof

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  1. J R Viola
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Correspondence to J R Viola, R Strömberg or C I E Smith.

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Supplementary Information accompanies the paper on Gene Therapy website (http://www.nature.com/gt)

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Viola, J., Leijonmarck, H., Simonson, O. et al. Fatty acid–spermine conjugates as DNA carriers for nonviral in vivo gene delivery. Gene Ther 16, 1429–1440 (2009). https://doi.org/10.1038/gt.2009.108

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