Potential therapeutic applications of recombinant, invasive E. coli

Critchley, R J; Jezzard, S; Radford, Kristen J.; Goussard, Sylvie; Lemoine, Nicholas R.; Grillot‐Courvalin, Catherine; Vassaux, Georges

doi:10.1038/sj.gt.3302281

Cited by 69 publications

(64 citation statements)

References 37 publications

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“…Furthermore, E. coli BM2710/pGB2Oinv-hly has recently been used to mediate in situ proteins delivery in tumor implanted in mice and the functional protein resulted in therapeutic benefits, when associated with systemic prodrugs delivery. 40 Although we did not directly characterize the cell population(s) expressing the transgene in vivo in normal animals, E. coli BM2710/pGB2Oinv-hly clearly invaded and remained confined to the mucosal epithelial layer driving the expression of the transgene throughout the gastrointestinal tract (Figure 4). However during colitis, taking into account the widespread distribution of plasmid-specific transcripts, it is possible that E. coli BM2710/pGB2Oinv-hly transferred the therapeutic gene to different cell populations (eg, phagocytic and endothelial cells) in extra-intestinal districts such as liver and lung, Figure 4).…”

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confidence: 96%

Engineered E. coli delivers therapeutic genes to the colonic mucosa

et al. 2005

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Taking advantage of the proximity of bowel mucosa to luminal bacteria, we have attempetd 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 pGB2Oinv-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 (pC1OTGF-b1) 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 pC1OTGF-b1 by the inflammation-inducible interleukin-8 promoter generating plasmid pC1OTGF-b1 IND . Plasmid-specific TGF-b1 mRNA transcripts were detectable in mouse CMT-93 epithelial cells incubated with E. coli BM2710/pGB2Oinv-hly carrying pC1OTGF-b1 IND 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/ pGB2Oinv-hly+pC1OTGF-b1. 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.

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confidence: 96%

Engineered E. coli delivers therapeutic genes to the colonic mucosa

et al. 2005

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“…Specific receptor clustering seems to be required for other bacterial-ligand receptor interactions (4, 10); therefore, our strategy has the potential to apply to the functional analysis for other bacterial entries. Furthermore, our system is able to be adapted to molecular delivery to mammalian epithelial cells expressing integrin β1, which is a recent topic of therapeutic utilization of bacterial proteins (5,6,8,9). INTRACELLULAR …”

Section: Discussionmentioning

confidence: 99%

“…Cells (2ϫ10 5 per well) were seeded in 6-well culture dishes (Corning, Acton, Mass., U.S.A.) and cultured 2 days in DMEM/F12 containing 10% FBS, which was then replaced by 1 ml of DMEM/F12 containing 3% BSA and 1.6 µg of monovalent or polyvalent 125 I-invasin. After being incubated for the indicated times, cells were washed four times with PBS containing 1% BSA and then lysed with RIPA buffer.…”

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Observation of the Intracellular Behavior of Recombinant Yersinia pseudotuberculosis Invasin Protein

Koga

Izawa

Araki

et al. 2005

Microbiology and Immunology

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In this study, we observed the intracellular behavior of recombinant invasin, a 103‐kDa outer membrane protein of Yersinia pseudotuberculosis. To mimic the in vivo behavior of bacterial invasin, a polyvalent form of invasin was generated by incubation of biotinylated GST‐fused invasin C‐terminal portion protein (GST‐INVS) with avidin. Several experiments confirmed that the recombinant invasin could consistently reproduce the invasin‐mediated entry to mammalian epithelial cells. We analyzed the molecular kinetics of polyvalent INVS by western blotting, 125I‐uptake, and immunofluorescent microscopy. The internalized polyvalent INVS was rapidly translocated to the RIPA‐insoluble (polymerized‐actin enriched) fraction and formed cytoplasmic vesicles, while monovalent invasin did not show such kinetics. From these observations, we concluded that our bacterial‐free system is able to analyze the action of invasin for Yersinia pseudotuberculosis entry.

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“…21 However, bacteria at high multiplicity of infection (MOI) may be toxic to mammalian cells. 22,23 In the present study, because A549 cells were more resistant to S. choleraesuis-induced cytotoxicity than B16F10 cells, and thereby less prone to detachment from culture plates after bacterial infection, we used them to demonstrate S. choleraesuis-mediated gene transfer in vitro. After an 8-hour incubation with S. choleraesuis at the initial inoculum level of approximately 10 MOIs, most, if not all, of the A549 cells remained intact.…”

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confidence: 99%

Systemic administration of attenuated Salmonella choleraesuis carrying thrombospondin-1 gene leads to tumor-specific transgene expression, delayed tumor growth and prolonged survival in the murine melanoma model

Lee

Shiau

2004

Cancer Gene Ther

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Some anaerobic and facultative anaerobic bacteria have been used experimentally as anticancer agents because of their selective growth in the hypoxia regions of solid tumors after systemic administration. We have previously shown the feasibility of using attenuated Salmonella choleraesuis as a gene delivery vector. In this study, we exploited S. choleraesuis carrying thrombospondin-1 (TSP-1) gene for treating primary melanoma and experimental pulmonary metastasis in the syngeneic murine B16F10 melanoma model. Systemic administration of S. choleraesuis allowed targeted gene delivery to tumors. The bacteria accumulated preferentially in tumors over livers and spleens at ratios ranging from 1000:1 to 10,000:1. The level of transgene expression via S. choleraesuis-mediated gene transfer in tumors could reach more than 1800-fold higher than in livers and spleens. Notably, bacterial accumulation was also observed in the lungs with metastatic nodules, but not in healthy lungs. When administered into mice bearing subcutaneous or pulmonary metastatic melanomas, S. choleraesuis carrying TSP-1 gene significantly inhibited tumor growth and enhanced survival of the mice. Immunohistochemical studies in the tumors from these mice displayed decreased intratumoral microvessel density. Taken together, these findings suggest that TSP-1 gene therapy delivered by S. choleraesuis may be effective for the treatment of primary as well as metastatic melanomas.

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Potential therapeutic applications of recombinant, invasive E. coli

Cited by 69 publications

References 37 publications

Engineered E. coli delivers therapeutic genes to the colonic mucosa

Engineered E. coli delivers therapeutic genes to the colonic mucosa

Observation of the Intracellular Behavior of Recombinant Yersinia pseudotuberculosis Invasin Protein

Systemic administration of attenuated Salmonella choleraesuis carrying thrombospondin-1 gene leads to tumor-specific transgene expression, delayed tumor growth and prolonged survival in the murine melanoma model

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