Gene delivery with optimized electroporation parameters shows potential for treatment of gliomas.

Yoshizato, Kimio; Nishi, Toru; Goto, Takehiko; Dev, Sukhendu B.; Takeshima, Hideo; Kino, Takeshi; Tada, Kohei; Kimura, Tooru; Shiraishi, Shoji; Kochi, Masato; Kuratsu, Jun Ichi; Hofmann, G.; Ushio, Yukitaka

doi:10.3892/ijo.16.5.899

Cited by 19 publications

(25 citation statements)

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“…This method has been used to introduce DNA into mouse skin (Titomirov et al, 1991), chicken embryos (Muramatsu et al, 1997), rat liver (Heller et al, 1996;Suzuki et al, 1998), mouse tumors (Rols et al, 1998;Goto et al, 2000), rat brain (Nishi et al, 1996), and rat corneal epithelium (Oshima et al, 1998). We and others have in addition demonstrated that the efficiency of transduction on plasmid delivery can be dramatically increased by applying an electric field to the muscle fibers subsequent to local DNA injection (Aihara and Miyazaki, 1998;Mir et al, 1998Mir et al, , 1999Mathiesen, 1999;Rizzuto et al, 1999;Vicat et al, 2000).…”

Section: Introductionmentioning

confidence: 91%

“…Electrical parameters, as well as appropriate electrodes, are likely to contribute to enhanced EGT. In particular, variations of voltage, duration of pulse, and rotation of the electric field have resulted in increased EGT of solid tumors (Yoshizato et al, 2000). Last, an in-depth analysis of the structural characteristics of the injected plasmid, ranging from the choice of a suitable promoter to the appropriate untranslated regions, will probably contribute to the establishment of efficient and long-lasting gene expression (Montgomery et al, 1993;Hartikka et al, 1996).…”

Section: Figmentioning

confidence: 99%

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Hyaluronidase Increases Electrogene Transfer Efficiency in Skeletal Muscle

et al. 2002

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Electrogene transfer (EGT) of plasmid DNA into skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic secretion of therapeutic proteins. We report here that preinjecting hyaluronidase (HYAse) significantly increases the gene transfer efficiency of muscle EGT. Three constructs encoding mouse erythropoietin (pCMV/mEPO), secreted alkaline phosphatase (pCMV/SeAP), and luciferase (pGGluc) were electroinjected intramuscularly in BALB/c mice and rabbits with and without HYAse pretreatment. Preinjection 1 or 4 hr before EGT increased EPO gene expression by about 5-fold in mice and maintained higher gene expression than plasmid EGT alone. A similar increment in gene expression was observed on pretreatment with HYAse and electroinjection of pCMV/mEPO into rabbit tibialis muscle. The increment of gene expression in rabbits reached 17-fold on injection of plasmid pCMV/SeAP and 24-fold with plasmid pGGluc. Injection of a plasmid encoding beta-galactosidase (pCMV/beta gal/NLS) and subsequent staining with 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside indicated that HYAse increased the tissue area involved in gene expression. No irreversible tissue damage was observed on histological analysis of treated muscles. HYAse is used in a variety of clinical applications, and thus the combination of HYAse pretreatment and muscle EGT may constitute an efficient gene transfer method to achieve therapeutic levels of gene expression.

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Section: Introductionmentioning

confidence: 91%

Section: Figmentioning

confidence: 99%

Hyaluronidase Increases Electrogene Transfer Efficiency in Skeletal Muscle

et al. 2002

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“…3 Marker genes were successfully transferred into normal tissues of animals including skeletal muscle, 4,5 liver, 6 skin 7 and cornea. 8 Tumors implanted in animals were also revealed to be good targets for in vivo electroporation; melanoma, 9 hepatocellular carcinoma, 10 and glioma 11 resulting in remarkable therapeutic outcome. The herpes simplex virus-type I thymidine kinase gene was transferred into tumors derived from the CT26 colon carcinoma cell line.…”

Section: Introductionmentioning

confidence: 99%

In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice

et al. 2001

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“…Recent reports have demonstrated optimized electroporation conditions including voltage, pulse duration, number of pulse, frequency, DNA concentration, and the amount of injected reporter gene in the tibialis anterior muscle (Golzio et al, 1998;Muramatsu et al, 1998;Gehl and Mir, 1999;Hofmann et al, 1999;Mir et al, 1999;Mekid and Mir, 2000;Miklavcic et al, 2000;Yoshizato et al, 2000;Lucas and Heller, 2001). It was also shown that square-wave electric pulses improve pulsed DNA delivery to living tissues .…”

Section: Introductionmentioning

confidence: 99%

Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation

Lee

Cho²,

Jang³

et al. 2002

Exp Mol Med

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In vivo electroporation has emerged as a leading technology for developing nonviral gene therapies, and the various technical parameters governing electroporation efficiency have been optimized by both theoretical and experimental analysis. However, most electroporation parameters focused on the electric conditions and the preferred vehicle for plasmid DNA injections has been normal saline. We hypothesized that salts in vehicle for plasmid DNA must affect the efficiency of DNA transfer because cations would alter ionic atmosphere, ionic strength, and conductivity of their medium. Here, we show that half saline (71 mM) is an optimal vehicle for in vivo electroporation of naked DNA in skeletal muscle. With various salt concentrations, two reporter genes, luciferase and β β β β-galactosidase were injected intramuscularly under our optimal electric condition (125 V/cm, 4 pulses x 2 times, 50 ms, 1 Hz). Exact salt concentrations of DNA vehicle were measured by the inductively coupled plasma-atomic emission spectrometer (ICP-AES) and the conductivity change in the tissue induced by the salt in the medium was measured by Low-Frequency (LF) Impedance Analyzer. Luciferase expression increased as cation concentration of vehicle decreased and this result can be visualized by X-Gal staining. However, at lower salt concentration, transfection efficiency was diminished because the hypoosmotic stress and electrical injury by low conductivity induced myofiber damage. At optimal salt concentration (71 mM), we observed a 3-fold average increase in luciferase expression in comparison with the normal saline condition (p < 0.01). These results provide a valuable experimental parameter for in vivo gene therapy mediated by electroporation.

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Gene delivery with optimized electroporation parameters shows potential for treatment of gliomas.

Cited by 19 publications

References 0 publications

Hyaluronidase Increases Electrogene Transfer Efficiency in Skeletal Muscle

Hyaluronidase Increases Electrogene Transfer Efficiency in Skeletal Muscle

In vivo electroporation-mediated transfer of interleukin-12 and interleukin-18 genes induces significant antitumor effects against melanoma in mice

Optimal salt concentration of vehicle for plasmid DNA enhances gene transfer mediated by electroporation

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