Long-term production of erythropoietin after electroporation-mediated transfer of plasmid DNA into the muscles of normal and uremic rats

Maruyama, Haruhiko; Ataka, Ken; Gejyo, Fumitake; Higuchi, Noboru; Ito, Yumi; Hirahara, Hiroyuki; Imazeki, Ikuo; Hirata, Michinori; Ichikawa, Fumihiko; Neichi, Tomohiro; Kikuchi, Hiroshi; Sugawa, Makoto; Miyazaki, Jun‐ichi

doi:10.1038/sj.gt.3301412

Cited by 72 publications

(69 citation statements)

References 32 publications

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“…The peak serum Epo levels following the tail vein injection of 800 mg of pCAGGS-Epo were 100-fold greater than the levels reached following the muscle-targeted gene transfer of 400 mg of pCAGGS-Epo by in vivo electroporation. 28,29 In the present study, the peak serum vIL-10 levels following the tail vein injection of 800 mg of pCAGGS-vIL-10 were at least three-fold greater than the levels reached following intravenous gene transfer with an adenovirus vector, peaking at 165.47101 ng/ml 1 day after the gene transfer. 30 Thus, our previous and present studies demonstrate that gene transfer into the liver by the hydrodynamics-based transfection method is easily applicable to the rat, which is more than 10 times larger than the mouse.…”

Section: Discussioncontrasting

confidence: 42%

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

et al. 2003

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Gene therapy is expected to revolutionize the treatment of kidney diseases. Viral interleukin (vIL)-10 has a variety of immunomodulatory properties. We examined the applicability of vIL-10 gene transfer to the treatment of rats with crescentic glomerulonephritis, a T helper 1 (Th 1) predominant disease. To produce the disease, Wistar-Kyoto rats were injected with a rabbit polyclonal anti-rat glomerular basement membrane antibody. After 3 h, a large volume of plasmid DNA expressing vIL-10 (pCAGGS-vIL-10) solution was rapidly injected into the tail vein. pCAGGS solution was similarly injected into control rats (pCAGGS rats). We confirmed the presence of vector-derived vIL-10 mainly in the liver and observed high serum vIL-10 levels in pCAGGSvIL-10-injected rats. Compared with the pCAGGS rats, the pCAGGS-vIL-10 rats showed significant therapeutic effects: reduced frequency of crescent formation, decrease in the number of total cells, macrophages, and CD4 + T cells in the glomeruli, decrease in urine protein, and attenuation of kidney dysfunction. Using quantitative real-time polymerase chain reaction, we also observed that this model was Th1-predominant in the glomeruli and that the ratio of the transcripts of CD4, interferon-g, tumor necrosis factor-a, and monocyte chemotactic protein-1 to the transcripts of glucose-6-phosphate dehydrogenase in the glomeruli were all significantly lower in the pCAGGS-vIL-10 rats than in the pCAGGS rats. These results demonstrate that pCAGGS-vIL-10 gene transfer by hydrodynamics-based transfection suppresses crescentic glomerulonephritis.

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Section: Discussioncontrasting

confidence: 42%

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

et al. 2003

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“…Applications of this technique have been extended from treating muscular dystrophy to using muscle for systemic delivery of therapeutic proteins. [97][98][99][100][101] The mechanisms by which this method enhances transgene expression appear to be relatively simple. Electroporation is effective only when plasmid DNA is injected into the muscle prior to, not after, electroporation.…”

Section: Electroporationmentioning

confidence: 99%

Non-viral gene delivery in skeletal muscle: a protein factory

Lu¹,

Bou‐Gharios²,

Partridge³

2003

Gene Ther

168

109

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Ever since the publication of the first reports in 1990 using skeletal muscle as a direct target for expressing foreign transgenes, an avalanche of papers has identified a variety of proteins that can be synthesized and correctly processed by skeletal muscle. The impetus to the development of such applications is not only amelioration of muscle diseases, but also a range of therapeutic applications, from immunization to delivery of therapeutic proteins, such as clotting factors and hormones. Although the most efficient way of introducing transgenes into muscle fibres has been by a variety of recombinant viral vectors, there are potential benefits in the use of non-viral vectors. In this review we assess the recent advances in construction and delivery of naked plasmid DNA to skeletal muscle and highlight the options available for further improvements to raise efficiency to therapeutic levels.

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“…Hematocrit and reticulocytes levels were determined as previously described. 14 serum Epo levels peaked at 35.5 ± 0.9 mU/ml at week 1, and gradually decreased with slight fluctuations; however, they remained more elevated at each time-point tested after DNA injection than did those of the rats given lower doses of the gene. Although skin-targeted gene transfer is less efficient than muscle-targeted gene transfer, these data indicate that skin-targeted gene transfer using in vivo electroporation can elicit the continuous delivery of Epo for 7 weeks in rats.…”

Section: Figure 5 Effects Of Epo Gene Transfer By Intradermal Electromentioning

confidence: 83%

“…Muscle-targeted gene transfer by in vivo electroporation was performed essentially as previously described. 14 A total of 200 g of plasmid DNA was injected into the lateral sides of both lower legs (100 g/each site). Hematocrit and reticulocytes levels were determined as previously described.…”

Section: Figure 5 Effects Of Epo Gene Transfer By Intradermal Electromentioning

confidence: 99%

Skin-targeted gene transfer using in vivo electroporation

et al. 2001

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The skin is an important target for gene transfer because of its easy accessibility. Using plasmid DNA expressing rat erythropoietin (pCAGGS-Epo) Because of its ready accessibility for direct manipulation and clinical monitoring, the skin is an attractive tissue for gene transfer. Direct in vivo plasmid DNA transfer to the skin via injection 1,2 and topical administration 3 have been reported. Intradermally injected plasmid DNA rapidly traverses the dermo-epidermal junction and is taken up and expressed by keratinocytes in the epidermis. 4 Active transgene expression in the epidermis is lost by 3 days after injection, 1 a time period that is too short for clinical applications. Moreover, skin takes up and expresses DNA less efficiently than does muscle. 5 Consequently, skin-targeted transfer of plasmid DNA has focused on two principal therapeutic uses: genetic immunization 6,7 and the expression of biological response modifiers to treat skin disease. 4 Gene gun 8,9 and jet injection 10 techniques have been developed for effective skin-targeted gene transfer. A gene gun technique has been used to successfully deliver naked DNA into the epidermis. 11 Klinman et al 11 also reported that, compared with muscle, skin is a suitable site for the short-term (3 weeks) production of erythropoietin (Epo), due to the rapid turnover of skin cells. Skin-targeted gene transfer by in vivo electroporation has been shown to be effective for introducing plasmid DNA, including the SV40 T antigen gene, into newborn mouse skin, 12 and the lacZ gene has been transferred into the hairless mouse with skin-depth targeting. 13 However, there are no reports on the systemic delivery of therapeutic proteins by skin-targeted gene transfer using electroporation. We demonstrate here that skin-targeted transfer of the Epo gene using electroporation with plate-and-fork-type electrodes produced physiologically significant levels of Epo in the systemic circulation for 7 weeks, and increased erythropoiesis in normal rats for 11 weeks.We tested three types of electrodes for skin-targeted gene transfer (Figure 1). To optimize the voltage of the electric pulses used for in vivo electroporation, we compared the serum Epo levels of rats subjected to electroporation at various voltages that did not cause macroscopic skin damage: 50 V, 24 V and 12 V, for the needletype electrodes, and 24 V, 18 V and 12 V, for the disktype and plate-and-fork-type electrodes. Pre-and postinjection serum Epo levels were measured on day Ϫ7 before and day 7 after intradermal injection with 800 g of either pCAGGS-Epo or pCAGGS. Only electroporation with the plate-and-fork-type electrodes at low voltages significantly increased the serum Epo levels (Figure 2). This result demonstrates that the type of electrode used is of critical importance in skin-targeted Epo gene transfer using in vivo electroporation. The difference in effectiveness of electrode types is probably explained by differences in the direction of the current to the skin. With the needle-type electrodes, the curre...

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Long-term production of erythropoietin after electroporation-mediated transfer of plasmid DNA into the muscles of normal and uremic rats

Cited by 72 publications

References 32 publications

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

Hydrodynamics-based delivery of the viral interleukin-10 gene suppresses experimental crescentic glomerulonephritis in Wistar–Kyoto rats

Non-viral gene delivery in skeletal muscle: a protein factory

Skin-targeted gene transfer using in vivo electroporation

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