Optimization of cutaneous electrically mediated plasmid DNA delivery using novel electrode

Heller, Loree C.; Jaroszeski, MJ; Coppola, Domenico; McCray, AN; Hickey, Joseph; Heller, Richard

doi:10.1038/sj.gt.3302867

Cited by 95 publications

(90 citation statements)

References 31 publications

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“…29,36 On the other hand, the combination of electroporation with i.d. delivery results in enhanced expression (unpublished results and Heller and Drabick 20,31 ), further implicating the involvement of membrane disruption in skin.…”

Section: Nuclei Are Visualized With Dapi (Blue)mentioning

confidence: 76%

“…47 Intradermal injection, with or without other enhancing methodologies such as electroporation, appears to be the most commonly used and most effective method identified to date to introduce nucleic acids into keratinocytes of the skin. 8,9,15,19,20,31 Previous studies using i.d. injection suggested that delivery of plasmid or siRNA across the stratum corneum barrier to the vicinity of the keratinocyte allowed uptake by unknown mechanisms.…”

Section: Discussionmentioning

confidence: 99%

“…injection [8][9][10][11] gene gun, 12,13 magnetofection, 14 electroporation [15][16][17][18][19][20] and topical formulations. [21][22][23][24][25][26] However, only limited success with sustainable gene expression has been observed over wide areas of the skin.…”

Section: Introductionmentioning

confidence: 99%

“…30 Although i.d. injection of nucleic acid appears to be the most efficient delivery technique for gene transfer to skin (particularly when combined with other technologies such as electroporation), 19,20,31 the localized and painful nature of hypodermic needle injection precludes its repeated use in treating wide areas of the skin. Therefore, development of novel, efficient and 'patientfriendly' delivery technologies is needed.…”

Section: Introductionmentioning

confidence: 99%

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Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

González-González

Spitler

et al. 2010

Gene Ther

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Nucleic acid-based therapies hold great promise for treatment of skin disorders if delivery challenges can be overcome. To investigate one mechanism of nucleic acid delivery to keratinocytes, a fixed mass of expression plasmid was intradermally injected into mouse footpads in different volumes, and reporter expression was monitored by intravital imaging or skin sectioning. Reporter gene expression increased with higher delivery volumes, suggesting that pressure drives nucleic acid uptake into cells after intradermal injections similar to previously published studies for muscle and liver. For spatiotemporal analysis of reporter gene expression, a dual-axis confocal (DAC) fluorescence microscope was used for intravital imaging following intradermal injections. Individual keratinocytes expressing hMGFP were readily visualized in vivo and initially appeared to preferentially express in the stratum granulosum and subsequently migrate to the stratum corneum over time. Fluorescence microscopy of frozen skin sections confirmed the patterns observed by intravital imaging. Intravital imaging with the DAC microscope is a noninvasive method for probing spatiotemporal control of gene expression and should facilitate development and testing of new nucleic acid delivery technologies.

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Section: Nuclei Are Visualized With Dapi (Blue)mentioning

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

González-González

Spitler

et al. 2010

Gene Ther

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“…The excellent immunogenicity of dermal DNA vaccination is probably related to the high prevalence of antigen-presenting cells (APCs) in the skin, in the form of Langerhans cells (LCs) in the epidermis and dendritic cells in the dermis (Kanitakis, 2002;Mathers and Larregina, 2006). Several techniques have been developed for the intradermal administration and cellular uptake of naked DNA, such as gene gun and particle injection systems (Klinman et al, 1998;Mitragotri, 2005;Steitz et al, 2006); jet injectors (Mitragotri, 2005;Bahloul et al, 2006); electroporators (Maruyama et al, 2001;Heller et al, 2007;Hooper et al, 2007;Hirao et al, 2008); and a technique, developed at our institute, that has been called ''DNA tattooing'' (Bins et al, 2005;Pokorna et al, 2008). DNA tattooing delivers naked plasmid DNA into the skin through thousands of punctures made with a multiple-needle tattoo device.…”

mentioning

confidence: 99%

Optimization of Intradermal Vaccination by DNA Tattooing in Human Skin

et al. 2009

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The intradermal administration of DNA vaccines by tattooing is a promising delivery technique for genetic immunization, with proven high immunogenicity in mice and in nonhuman primates. However, the parameters that result in optimal expression of DNA vaccines that are applied by this strategy to human skin are currently unknown. To address this issue we set up an ex vivo human skin model in which DNA vaccine-induced expression of reporter proteins could be monitored longitudinally. Using this model we demonstrate the following: First, the vast majority of cells that express DNA vaccine-encoded antigen in human skin are formed by epidermal keratinocytes, with only a small fraction (about 1%) of antigen-positive epidermal Langerhans cells. Second, using full randomization of DNA tattoo variables we show that an increase in DNA concentration, needle depth, and tattoo time all significantly increase antigen expression ( p < 0.001), with DNA concentration forming the most critical variable influencing the level of antigen expression. Finally, in spite of the marked immunogenicity of this vaccination method in animal models, transfection efficiency of the technique is shown to be extremely low, estimated at approximately 2 to 2000 out of 1Â10 10 copies of plasmid applied. This finding, coupled with the observed dependency of antigen expression on DNA concentration, suggests that the development of strategies that can enhance in vivo transfection efficacy would be highly valuable. Collectively, this study shows that an ex vivo human skin model can be used to determine the factors that control vaccine-induced antigen expression and define the optimal parameters for the evaluation of DNA tattoo or other dermal delivery techniques in phase 1 clinical trials.

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Transfection by Electroporation

Potter

Heller

2011

CP Neuroscience

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Electroporation--the use of high-voltage electric shocks to introduce DNA into cells--can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit describes electroporation of mammalian cells, including ES cells for the preparation of knock-out, knock-in, and transgenic mice. Protocols are described for the use of electroporation in vivo to perform gene therapy for cancer therapy and DNA vaccination. Also described are modifications for preparation and transfection of plant protoplasts.

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Optimization of cutaneous electrically mediated plasmid DNA delivery using novel electrode

Cited by 95 publications

References 31 publications

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

Increased interstitial pressure improves nucleic acid delivery to skin enabling a comparative analysis of constitutive promoters

Optimization of Intradermal Vaccination by DNA Tattooing in Human Skin

Transfection by Electroporation

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