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Formation of LID vector complexes in water alters physicochemical properties and enhances pulmonary gene expression in vivo

Gene Therapy volume 10, pages 1026–1034 (2003)Cite this article

Abstract

There is currently an urgent need to develop efficient gene-delivery systems for the lung that are free of inflammatory effects. The LID vector is a synthetic gene delivery system, comprised of lipofectin (L), an integrin-targeting peptide (I) and DNA (D) that has previously been shown to have high transfection efficiency in the lung. We have assessed the effect of alternative methods of complex preparation on structural features of the complex, levels and duration of reporter gene expression and the host response to the LID vector. We have demonstrated that making the complex in water affects the structure of the LID complexes making them smaller and more stable with a more cationic surface charge than complexes prepared in phosphate-buffered saline (PBS). When the LID vector was constituted in water and instilled intratracheally into the lungs of mice there was a 10-fold increase in luciferase activity compared with preparation in PBS. Furthermore, luciferase activity was still evident 1 week following vector instillation. This enhancement may be because of altered complex structure, although effects of the hypotonic vector solution on the lung cannot be excluded. The inflammatory effects of instilling the LID vector in water were minimal, even after three administrations of the LID vector, with only mild alterations in cytokine and broncho-alveolar lavage fluid (BALF) cell profiles. These results demonstrate that the LID vector can generate high, and prolonged, levels of gene expression in the lung from small quantities of DNA and that careful attention to synthetic polyplex structure may be important to optimize efficiency of gene expression in vivo.

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Acknowledgements

The liposome DOTMA/DOPE in a 1:1 molar ratio was generously provided by Valentis Inc. (Burlingame, CA, USA). RGJ is an ARC Clinical Research Fellow. The study was supported by the Arthritis Research Campaign, the Wellcome Trust, and the British Lung Foundation.

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Author notes

  1. The last two authors contributed equally to this study

Authors and Affiliations

  1. Centre for Cardiopulmonary Biochemistry and Respiratory Medicine, Royal Free and University College Medical School, University College London, Rayne Institute, London, UK

    R G Jenkins, R J Hodges, S E W Bottoms, G J Laurent & R J McAnulty

  2. Molecular Immunology Unit, Institute of Child Health, University College London, London, UK

    Q-H Meng & S L Hart

  3. Department of Biomechanical Engineering, University College London, Torrington Place, London, UK

    L K Lee & P Ayazi Shamlou

  4. Department of Pharmacology, Royal Free and University College Medical School, London, UK

    D Willis

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  1. R G Jenkins
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  10. S L Hart
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Jenkins, R., Meng, QH., Hodges, R. et al. Formation of LID vector complexes in water alters physicochemical properties and enhances pulmonary gene expression in vivo. Gene Ther 10, 1026–1034 (2003). https://doi.org/10.1038/sj.gt.3301963

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