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Potential therapeutic applications of recombinant, invasive E. coli

Abstract

An invasive Escherichia coli expressing the inv gene from Yersinia pseudotuberculosis was used as a vector for protein delivery to mammalian epithelial cells. Upon incubation with β1-integrin-expressing mammalian cells, the bacteria are internalized, allowing bacteria-encoded proteins to function from within the mammalian cell. These bacteria are eventually processed in the host phagosome where they are destroyed. Expression of listeriolysin O from Listeria monocytogenes in the bacterium and its subsequent release into the phagosome triggers the breakdown of the membrane, allowing the release of the bacterial content into the cytosol of host cells. Using this vector, we demonstrate delivery of a gene and intact, functional proteins into mammalian cells in which β1-integrin is expressed and accessible. At a ratio of bacteria/mammalian cells compatible with the survival of the mammalian cells, protein delivery can be observed in the entire cell population in vitro, while gene transfer is far less efficient. Protein delivery can also be achieved in vivo in mouse tumour models and can be detected at least 96 h after inoculation. Functional, natural E. coli proteins are delivered in the process and can provide therapeutic benefit in vivo, when associated with prodrugs. This therapeutic effect is associated with infiltration of neutrophils, eosinophils, macrophages and to a lesser extent dendritic cells in the tumour mass.

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Acknowledgements

We thank Dr GW Both (University of New South Wales, Australia) for providing 6-MPDR used in this study. Panc-O2 cells were kindly provided by Dr Hollingsworth (University of Nebraska Medical Center, Omaha, USA) and the cell line GD25 and its β1-integrin-expressing counterpart were provided by Dr R Fassler (Max Planck-Institute for Biochemistry, Germany). Special thanks are due to Dr Y Wang for his help with IHC staining analysis. This work was supported by Cancer Research UK and ‘Help Hammer Cancer’.

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Critchley, R., Jezzard, S., Radford, K. et al. Potential therapeutic applications of recombinant, invasive E. coli. Gene Ther 11, 1224–1233 (2004). https://doi.org/10.1038/sj.gt.3302281

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