Long-term persistence of plasmid DNA and foreign gone expression in mouse muscle

Wolff, Jon A.; Ludtke, James J.; Acsádi, Gyula; Williams, Philip M.; Jáni, Ágnes

doi:10.1093/hmg/1.6.363

Cited by 703 publications

(343 citation statements)

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“…28 In another study, plasmid DNA persisted for at least 19 months after injection into skeletal muscle. 29 The PCR assay is a highly efficient gene amplification procedure that has been increasingly applied to the safety assessment of gene therapy. However, there have been no consistent requirements for assay sensitivity in the development of intratumoral gene therapy approaches.…”

Section: Discussionmentioning

confidence: 99%

Tissue distribution of liposome-mediated epidermal growth factor receptor antisense gene therapy

et al. 2003

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Despite the widespread use of liposome-mediated gene transfer in cancer therapy protocols, little is known about the tissue distribution of intralesionally administered DNA. We have previously shown that antisense gene therapy targeting the epidermal growth factor receptor (EGFR) inhibited tumor growth in a human head and neck squamous cell carcinoma (HNSCC) xenograft model. Further investigation demonstrated lack of systemic toxicity with intramuscular or intratumoral administration of this liposomal-DNA complex. In the present study, we compared two approaches to determine the presence of exogenous DNA in the plasma and tissues of mice treated with intramuscular injection of EGFR antisense gene therapy. PCR analysis using genomic DNA plus plasmid DNA as template was 83-fold more sensitive than PCR using a mixture of total RNA and plasmid DNA as template. With the more sensitive method (able to detect fewer than 500 molecules of EGFR antisense DNA in 1 mg of genomic DNA), foreign DNA was detected in all organs up to 1 month following a single injection. In contrast, using RNA plus plasmid DNA as template, exogenous DNA was only detected at the injection site at 1 week, and was undetectable at 1 month. Optical imaging studies demonstrated plasmid DNA only at the injection site. Although less sensitive than PCCR, Southern blot hybridization showed no evidence of integration of foreign DNA into the host genome in vitro or in vivo. These results emphasize the importance of defining the assays used to detect foreign DNA and suggest that the ability to detect intralesionally administered liposomal gene therapy, in organs distant from the injection site, is directly correlated with the sensitivity of the method employed.

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Section: Discussionmentioning

confidence: 99%

Tissue distribution of liposome-mediated epidermal growth factor receptor antisense gene therapy

et al. 2003

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“…With the exception of intramuscular injection of plasmid DNA, 29 it has been postulated that naked DNA, once entering the target cells either by nonviral vector-mediated procedure or other physical methods, is quickly degraded, 28 The kinetic pattern of hAAT in serum shown in Figure 3 probably results from an equilibrium between the rate of hAAT elimination and the rate of production by the transfected liver cells. Considering that the blood halflife of hAAT in mouse is about 15 h, 33 which has been confirmed by our own measurements, it is likely that hAAT production dominates the serum level at an early time-point when the amount of hAAT gene in the liver is maximal.…”

Section: Discussionmentioning

confidence: 99%

Long-term expression of human alpha1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure

2000

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The liver is an important target organ for gene transfer due to its large capacity for synthesizing serum proteins and its involvement in numerous genetic and acquired diseases. Previously, we and others have shown that an efficient gene transfer to liver cells in vivo can be achieved by an intravenous injection of plasmid DNA using a hydrodynamics-based procedure. In this study, we systematically characterized the expression of transgene encoding a secretory protein in mouse. Using human ␣1-antitrypsin (hAAT) gene as a reporter, we demonstrate that the serum level of hAAT can reach as high as 0.5 mg/ml by a simple

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“…27 ± 29 However, plasmid DNA has been also shown to persist for as long as 19 months. 29 We could detect the plasmid for at least 3 months after injection, and during this period, the plasmids were actively transcribed ( Fig 7) . Translation of the plasmid into protein also took place for at least 7 days (Fig 8 ), a time interval adequate for invoking humoral and cellular immunity in mice.…”

Section: Discussionmentioning

confidence: 87%

Construction and characterization of DNA vaccines encoding the single-chain variable fragment of the anti-idiotype antibody 1A7 mimicking the tumor-associated antigen disialoganglioside GD2

Zeytin

Tripathi

Bhattacharya‐Chatterjee

et al. 2000

Cancer Gene Ther

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Anti -idiotype antibody, 1A7, functionally mimics the tumor -associated antigen disialoganglioside GD2, which is overexpressed on the surface of a number of neuroectodermal tumors such as melanoma, neuroblastoma, soft tissue sarcoma, and small cell carcinoma of the lung. Immunization of mice with 1A7 generated the production of anti -GD2 antibodies. In a phase I clinical trial, immunization of patients with 1A7, mixed with the adjuvant QS21, demonstrated that 1A7 could act as a surrogate antigen for GD2 and induce strong humoral immune responses in advanced stage melanoma patients. DNA vaccines have recently been shown to invoke humoral as well as cellular responses in injected hosts against the transgene product. To evaluate the efficiency of DNA vaccines encoding anti -idiotype antibodies, we constructed expression plasmids encoding the variable heavy ( VH ) and variable light ( VL ) chains of 1A7. The plasmids were made in two configurations, expressing either the VH ( pc1A7VHLnVL ) or the VL ( pc1A7VLLnVH ) chain of 1A7 at the amino terminus, linked together by a 15 -amino acid linker ( Ln ) . In vitro transcription / translation assays and transfection of CHO -K1 cells with the plasmids demonstrated that a $30 -kDa protein was expressed by both configurations of the single -chain variable fragment. This protein can be specifically precipitated by monoclonal anti -GD2 antibody, 14G2a. Following intramuscular injection in mice, the plasmids were detectable in the injected tissues for at least 3 months and the injected plasmids actively transcribed the single -chain variable fragment 1A7 gene at the injected site. A single, intramuscular immunization of a group of C57BL / 6 mice with pc1A7VLLnVH in phosphate -buffered saline induced humoral immune responses against 1A7 as well as GD2, the nominal antigen. Multiple immunizations, however, were required to elicit stronger immune responses.

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Long-term persistence of plasmid DNA and foreign gone expression in mouse muscle

Cited by 703 publications

References 0 publications

Tissue distribution of liposome-mediated epidermal growth factor receptor antisense gene therapy

Tissue distribution of liposome-mediated epidermal growth factor receptor antisense gene therapy

Long-term expression of human alpha1-antitrypsin gene in mouse liver achieved by intravenous administration of plasmid DNA using a hydrodynamics-based procedure

Construction and characterization of DNA vaccines encoding the single-chain variable fragment of the anti-idiotype antibody 1A7 mimicking the tumor-associated antigen disialoganglioside GD2

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