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A multifunctional PEI-based cationic polyplex for enhanced systemic p53-mediated gene therapy

Gene Therapy volume 13, pages 1512–1523 (2006)Cite this article

Abstract

We recently reported a novel coupling strategy involving salicylhydroxamic acid and phenyl(di)boronic acid molecules to attach the CNGRC peptide to PEI/DNA for CD13 targeting in tumors. Here, we doubly coupled Simian Virus (SV) 40 peptide-(nuclear localization signal)) and oligonucleotide-based (DNA nuclear targeting signal) nuclear signals to the same vector using peptide nucleic acid chemistry. This vector, CNGRC/PEG/PEI/DNA-βgal/NLS/DNTS, was predominantly localized in the cell nucleus, yielding about 200-fold higher βgal gene expression in vitro, more than 20-fold increase in tumor-specific gene delivery, and a robust βgal gene expression as demonstrated in stained tumor sections. For gene therapy purposes, we further engineered a similar targeting polyplex, CNGRC/PEG/PEI/DNA-p53/NLS/DNTS, with EBV-based episomal vector for sustained p53 gene expression. A distribution of vector DNA and apoptosis in p53-containing tumors was observed, yielding a significant tumor regression and 95% animal survival after 60 days. This multicomponent vector also co-targeted tumor and tumor-associated endothelial cells but not normal cells, and had more efficient therapeutic index than each vector administered as a single modality. The use of an efficient coupling strategy without compromising the vector's integrity for DNA condensation and endosomal escape; nuclear import; tumor-specific and persistent p53 gene expression clearly provides a basis for developing a single combinatorial approach for non-viral gene therapy.

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Acknowledgements

We are grateful to Dr Randall Evans (Department of Molecular Hematology and Therapy) of the Flow Cytometry/Cell Sorting and Confocal Microscope/Image Analysis Core of the University of Texas MD Anderson Cancer Center for his invaluable technical assistance in confocal microscopy. This work was supported in part by NIH grant #5P30CA016672–29 supporting the core facility.

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  1. Department of Genitourinary Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA

    S Moffatt, S Wiehle & R J Cristiano

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  1. S Moffatt
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Correspondence to S Moffatt.

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Moffatt, S., Wiehle, S. & Cristiano, R. A multifunctional PEI-based cationic polyplex for enhanced systemic p53-mediated gene therapy. Gene Ther 13, 1512–1523 (2006). https://doi.org/10.1038/sj.gt.3302773

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