Liposome-mediated, nonviral gene transfer induces a systemic inflammatory response which can exacerbate pre-existing inflammation

Norman, James; Denham, Woody; Denham, Daphne W.; Yang, Jie; Carter, G.; Abouhamze, Amer; Tannahill, Cynthia L.; MacKay, Sally L. D.; Moldawer, Lyle L.

doi:10.1038/sj.gt.3301240

Cited by 43 publications

(16 citation statements)

References 26 publications

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“…This is in accordance with data on pBAD expression plasmids for which no significant gene induction effects have been observed under similar low glucose conditions (26). By changing media type to modified M9 minimal media, glucose 3 J.-M. Collombet, unpublished. levels could be kept low (0.2%) and still effectively down-regulate cre expression using pMEV8, although this richer media type permitted increased plasmid yields over that of minimal media alone.…”

Section: Creation Of a Bacterial Strain Expressing Cre Recombinase Unsupporting

confidence: 75%

“…This expression construct is difficult to modify due to its instability (21) and presents problems for introduction into mitochondria by electroporation, due to its large size (28). 3 In addition, the bacterial vector falls within the mitochondrial gene COXIII, is not easily removable, and is likely to abolish mitochondrial gene function.…”

Section: Creation Of a Bacterial Strain Expressing Cre Recombinase Unmentioning

confidence: 99%

“…There is increasing evidence to suggest that plasmid DNA used for non-viral gene delivery can cause unacceptable inflammatory responses in eukaryotes (1)(2)(3)(4)(5). These immunotoxic responses are largely due to the presence of unmethylated CpG motifs and their associated stimulatory sequences on plasmids, following bacterial propagation of plasmid DNA.…”

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confidence: 99%

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An araC-controlled Bacterialcre Expression System to Produce DNA Minicircle Vectors for Nuclear and Mitochondrial Gene Therapy

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Tolmachov

Collombet

et al. 2001

Journal of Biological Chemistry

146

113

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The presence of CpG motifs and their associated sequences in bacterial DNA causes an immunotoxic response following the delivery of these plasmid vectors into mammalian hosts. We describe a biotechnological approach to the elimination of this problem by the creation of a bacterial cre recombinase expression system, tightly controlled by the arabinose regulon. This permits the Cre-mediated and -directed excision of the entire bacterial vector sequences from plasmid constructs to create supercoiled gene expression minicircles for gene therapy. Minicircle yields using standard culture volumes are sufficient for most in vitro and in vivo applications whereas minicircle expression in vitro is significantly increased over standard plasmid transfection. By the simple expedient of removing the bacterial DNA complement, we significantly reduce the size and CpG content of these expression vectors, which should also reduce DNA-induced inflammatory responses in a dose-dependent manner. We further describe the generation of minicircle expression vectors for mammalian mitochondrial gene therapy, for which no other vector systems currently exist. The removal of bacterial vector sequences should permit appropriate transcription and correct transcriptional cleavage from the mitochondrial minicircle constructs in a mitochondrial environment and brings the realization of mitochondrial gene therapy a step closer.There is increasing evidence to suggest that plasmid DNA used for non-viral gene delivery can cause unacceptable inflammatory responses in eukaryotes (1-5). These immunotoxic responses are largely due to the presence of unmethylated CpG motifs and their associated stimulatory sequences on plasmids, following bacterial propagation of plasmid DNA. Simple methylation of DNA in vitro may be enough to reduce an inflammatory response but is likely to result in severely depressed gene expression (6). The removal of CpG islands by cloning out, or elimination of non-essential sequences is more successful in reducing inflammatory responses but is time-consuming and tedious (7).Because bacterial DNA contains on average four times more CpG islands than does mammalian DNA (8), a good solution is to entirely eliminate the bacterial control regions from gene delivery vectors during the process of plasmid production. Removal of bacterial sequences needs to be efficient, using the smallest possible excision site, while creating supercoiled DNA minicircles, consisting solely of gene expression elements under appropriate mammalian control regions.This can be achieved by the use of Cre recombinase, a bacteriophage P1-derived integrase (9 -11), catalyzing site-specific recombination between direct repeats of 34 base pairs (loxP sites).In the case of a supercoiled plasmid containing DNA flanked by two loxP sites in the same orientation, Cre recombination produces two DNA molecules that are topologically unlinked, circular, and mainly supercoiled (10), each containing a single 34-bp 1 loxP site. Efficient minicircle production requires the use of...

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Section: Creation Of a Bacterial Strain Expressing Cre Recombinase Unsupporting

confidence: 75%

Section: Creation Of a Bacterial Strain Expressing Cre Recombinase Unmentioning

confidence: 99%

See 1 more Smart Citation

An araC-controlled Bacterialcre Expression System to Produce DNA Minicircle Vectors for Nuclear and Mitochondrial Gene Therapy

Bigger

Tolmachov

Collombet

et al. 2001

Journal of Biological Chemistry

146

113

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“…3), whereas we saw no evidence of lung inflammation in mice after CDE-induced pancreatitis, as determined histologically or by lung myeloperoxidase content. In previous studies with the CDE model, we observed increased expression of the proinflammatory cytokines TNF-␣ and IL-1 not only in the lung, but also in the liver and pancreas (29). Because Ad has such a propensity for the liver, and to a lesser extent, the pancreas, we repeated the studies with the i.v.…”

Section: Discussionmentioning

confidence: 99%

Adenoviral Delivery of Human and Viral IL-10 in Murine Sepsis

Minter

Ferry

Murday

et al. 2001

The Journal of Immunology

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Adenovirus (Ad) gene therapy has been proposed as a drug-delivery system for the targeted administration of protein-based therapies, including growth factors and biological response modifiers. However, inflammation associated with Ad transduction has raised concern about its safety and efficacy in acute inflammatory diseases. In the present report, intratracheal and i.v. administration of a first-generation adenoviral recombinant (E1,E3 deleted) either containing an empty cassette or expressing the anti-inflammatory cytokines viral or human IL-10 (IL-10) was administered to mice subjected to zymosan-induced multisystem organ failure or to acute necrotizing pancreatitis. Pretreatment of mice with the intratracheal instillation of Ad expressing human IL-10 or viral IL-10 reduced weight loss, attenuated the proinflammatory cytokine response, and reduced mortality in the zymosan-induced model, whereas pretreatment with a control adenoviral recombinant did not significantly exacerbate the response. Pretreatment of mice with pancreatitis using adenoviral vectors expressing IL-10 significantly reduced the degree of pancreatic and liver injury and liver inflammation when administered systemically, but not intratracheally. We conclude that adenoviral vectors can be administered prophylactically in acute inflammatory syndromes, and expression of the anti-inflammatory protein IL-10 can be used to suppress the underlying inflammatory process.

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“…While the inflammatory response to adenoviral vectors is well described, both plasmid DNA and liposomal complexes also potentially induce inflammation. 8,9 Despite the ease of production and scale-up of plasmid and liposomal complexes, low transmission efficiency and transgene expression are limiting. Transmission efficiency of plasmid DNA may be improved by the use of ultrasound.…”

Section: Viral and Nonviral Gene Transfer Agents Have Been Successfulmentioning

confidence: 99%

Gene therapy progress and prospects: therapeutic angiogenesis for limb and myocardial ischemia

2003

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Liposome-mediated, nonviral gene transfer induces a systemic inflammatory response which can exacerbate pre-existing inflammation

Cited by 43 publications

References 26 publications

An araC-controlled Bacterialcre Expression System to Produce DNA Minicircle Vectors for Nuclear and Mitochondrial Gene Therapy

An araC-controlled Bacterialcre Expression System to Produce DNA Minicircle Vectors for Nuclear and Mitochondrial Gene Therapy

Adenoviral Delivery of Human and Viral IL-10 in Murine Sepsis

Gene therapy progress and prospects: therapeutic angiogenesis for limb and myocardial ischemia

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