Engineered E. coli delivers therapeutic genes to the colonic mucosa
| Autor: | Ignazio Castagliuolo, Paola Brun, Elisa Beggiao, Catherine Grillot-Courvalin, Sylvie Goussard, Riccardo Manganelli, Giorgio Palù, Luisa Barzon |
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| Rok vydání: | 2005 |
| Předmět: |
Colon
Genetic enhancement Bacterial Toxins Genetic Vectors Administration Oral Biology medicine.disease_cause Intestinal absorption Microbiology Transforming Growth Factor beta1 Hemolysin Proteins Mice Plasmid Intestinal mucosa Transforming Growth Factor beta Genetics medicine Animals Intestinal Mucosa Promoter Regions Genetic Molecular Biology Escherichia coli Heat-Shock Proteins Regulation of gene expression Escherichia coli Proteins Epithelial Cells Genetic Therapy Colitis Listeria monocytogenes Gene Expression Regulation Intestinal Absorption Perforin Yersinia pseudotuberculosis biology.protein Molecular Medicine Cytolysin Genetic Engineering |
| Zdroj: | Gene Therapy. 12:1070-1078 |
| ISSN: | 1476-5462 0969-7128 |
| Popis: | Taking advantage of the proximity of bowel mucosa to luminal bacteria, we have attempted to deliver a therapeutic gene to the colonic mucosa by oral administration of an invasive and non-pathogenic Escherichia coli. E. coli diamenopimelate (dap) auxotroph, harboring plasmid pGB2Omegainv-hly, express the inv gene from Yersinia pseudotubercolosis that confers the ability to invade nonprofessional phagocytic cells and the hly gene from Listeria monocytogenes that allows expression of lystreriolysin O, a perforin cytolysin able to perfore phagosomal membranes. This bacterial vector invades and transfers functional DNA to epithelial cells in vitro. We have shown that this strain carrying a therapeutic gene (pC1OmegaTGF-beta1) can significantly reduce the severity of experimental colitis in mice. However, as a consequence of mucosal barrier disruption during colitis, vector-specific mRNA transcripts could be recovered from the colon and also from extra-colonic tissues. We therefore replaced the constitutive CMV promoter in pC1OmegaTGF-beta1 by the inflammation-inducible interleukin-8 promoter generating plasmid pC1OmegaTGF-beta1IND. Plasmid-specific TGF-beta1 mRNA transcripts were detectable in mouse CMT-93 epithelial cells incubated with E. coli BM2710/pGB2Omegainv-hly carrying pC1OmegaTGF-beta1IND following exposure to inflammatory cytokines. Furthermore, the transcripts were detectable only within inflamed tissues and the therapeutic effects were comparable to those in animals treated with E. coli BM2710/pGB2Omegainv-hly+pC1OmegaTGF-beta1. In summary, engineered enteric bacteria can efficiently deliver in vivo therapeutic genes to the intact intestinal mucosa and regulation expression of the therapeutic gene by an inflammation-inducible promoter prevents its dissemination during colitis. |
| Databáze: | OpenAIRE |
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