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A model for the analysis of nonviral gene therapy

Gene Therapy volume 10, pages 1766–1775 (2003)Cite this article

Abstract

Further understanding of the mechanisms involved in cellular and intracellular delivery of transgene is needed to produce clinical applications of gene therapy. The compartmental and computational model designed in this work is integrated with data from previous experiments to quantitatively estimate rate constants of plasmid translocation across cellular barriers in transgene delivery in vitro. The experimental conditions between two cellular studies were held constant, varying only the cell type, to investigate how the rates differed between cell lines. Two rate constants were estimated per barrier for active transport and passive diffusion. Translocation rates of intact plasmid across the cytoplasmic and nuclear barriers varied between cell lines. CV1 cells were defined by slower rates (0.23 h−1 cytoplasmic, 0.08 h–1 nuclear) than those of the HeLa cells (1.87 h−1 cytoplasmic, 0.45 h−1 nuclear). The nuclear envelope was identified as a rate-limiting barrier by comparing the rate of intact plasmid translocation at each barrier. Slower intact plasmid translocation in CV1 cells was correlated with a reduced absolute capacity for transgene efficiency in comparison with HeLa cells. HeLa cells were three times more efficient than CV1 cells at producing green fluorescent protein per intact plasmid delivered to the nucleus. Mathematical modeling coordinated with experimental studies can provide detailed, quantitative understanding of nonviral gene therapy.

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Acknowledgements

This work was supported by the Bioengineering and Environmental Systems Division of the National Science Foundation under Award BES-9902697. We would also like to acknowledge MB James for use of her experimental data on nonviral gene delivery and Dr MI Miga for reviewing this work.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Vanderbilt University, Nashville, TN, USA

    G A Banks, R J Roselli, R Chen & T D Giorgio

  2. Department of Chemical Engineering, Vanderbilt University, Nashville, TN, USA

    T D Giorgio

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  1. G A Banks
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  2. R J Roselli
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  4. T D Giorgio
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Banks, G., Roselli, R., Chen, R. et al. A model for the analysis of nonviral gene therapy. Gene Ther 10, 1766–1775 (2003). https://doi.org/10.1038/sj.gt.3302076

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