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. 2011 May 10;151(3):256-62.
doi: 10.1016/j.jconrel.2011.01.014. Epub 2011 Jan 22.

Electro-gene transfer to skin using a noninvasive multielectrode array

Siqi Guo  1 , Amy Donate, Gaurav Basu, Cathryn Lundberg, Loree Heller, Richard Heller
Affiliations

Electro-gene transfer to skin using a noninvasive multielectrode array

Siqi Guo et al. J Control Release. .

Abstract

Because of its large surface area and easy access for both delivery and monitoring, the skin is an attractive target for gene therapy for cutaneous diseases, vaccinations and several metabolic disorders. The critical factors for DNA delivery to the skin by electroporation (EP) are effective expression levels and minimal or no tissue damage. Here, we evaluated the non-invasive multielectrode array (MEA) for gene electrotransfer. For these studies we utilized a guinea pig model, which has been shown to have a similar thickness and structure to human skin. Our results demonstrate significantly increased gene expression 2 to 3 logs above injection of plasmid DNA alone over 15 days. Furthermore, gene expression could be enhanced by increasing the size of the treatment area. Transgene-expressing cells were observed exclusively in the epidermal layer of the skin. In contrast to caliper or plate electrodes, skin EP with the MEA greatly reduced muscle twitching and resulted in minimal and completely recoverable skin damage. These results suggest that EP with MEA can be an efficient and non-invasive skin delivery method with less adverse side effects than other EP delivery systems and promising clinical applications.

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Conflict of interest statement

Conflict of interest

With respect to duality of interest and financial disclosures, Dr. R. Heller is an inventor on patents which cover the technology that was used in the work reported in this manuscript. In addition, Dr. R. Heller owns stock and stock options in Inovio Pharmaceutical Corporation and has an ownership interest in RMR Technologies.

Figures

Figure 1
Figure 1. Kinetic of gene expression in skin after i.d. DNA (gWiz-Luciferase) injection and non-invasive EP
Delivery groups, 50µL-IO: 50µL DNA without EP; 50µL-1EP: 50µL DNA with 1 EP on the injection site; 200µL-IO: 200µL DNA without EP; 200µL-1EP: 200µL DNA and 1 EP; 200µL-4EP: 200µL DNA and 4 EPs; 50µLx4-IO: 4 injections with 50µL DNA without EP; 50µLx4-4EP: 4 injections with 50µL DNA and each EP on the injection site. A, Time course of luciferase expression in guinea pig skin after 1 time delivery at d0. Bars represent mean ± SD. 4–5 sites were analyzed for each delivery. B, Time course of luciferase expression in guinea pig skin with 3 time deliverys, separately at d0, d15 and d29. Bars represent mean ± SD. 5–6 sites were analyzed for each delivery. p/s = photons/second.
Figure 1
Figure 1. Kinetic of gene expression in skin after i.d. DNA (gWiz-Luciferase) injection and non-invasive EP
Delivery groups, 50µL-IO: 50µL DNA without EP; 50µL-1EP: 50µL DNA with 1 EP on the injection site; 200µL-IO: 200µL DNA without EP; 200µL-1EP: 200µL DNA and 1 EP; 200µL-4EP: 200µL DNA and 4 EPs; 50µLx4-IO: 4 injections with 50µL DNA without EP; 50µLx4-4EP: 4 injections with 50µL DNA and each EP on the injection site. A, Time course of luciferase expression in guinea pig skin after 1 time delivery at d0. Bars represent mean ± SD. 4–5 sites were analyzed for each delivery. B, Time course of luciferase expression in guinea pig skin with 3 time deliverys, separately at d0, d15 and d29. Bars represent mean ± SD. 5–6 sites were analyzed for each delivery. p/s = photons/second.
Figure 2
Figure 2. Distribution of gene-expressing cells after i.d. DNA (gWiz-GFP) injection and non-invasive EP
Skin samples were collected post-delivery, 1hour, day 1, day 2, day 7 or day 9. Samples were analyzed by immunoflurescence microscopy. Delivery group, 50µL-IO: 50µL DNA without EP; 50µL-1EP: 50µL DNA with 1 EP on the injection site; 50µLx4-4EP: 4 injection of 50µL DNA and each EP on the injection site; 200µL-1EP: 200µL DNA and 1 EP; 200µL-4EP: 200µL DNA and 4 EPs. A, One representative picture of 3 treated sites. (B, C, D) Total 6 cryosections (2 sections per sample) of each delivery were analyzed. Cell nuclei were blue-stained by DAPI. GFP-expressing cells were shown green. (C, D) Cell nuclei and stratum corneum was shown red-stained by propidium iodide. B, One representative section of each delivery was presented for post-delivery day 2 and day 7 (magnification = 100, scale bar=100µm). C, One representative section from post-delivery day 2 (magnification = 200). D, One representative section from post-delivery day 7 (magnification = 200, scale bar=100µm).
Figure 3
Figure 3. Gross observation and histology of skin after i.d. DNA injection and non-invasive EP
A, Skin observation after delivery. Pictures were taken at post-delivery day 1, day 2 and day 5. One representative picture of 4 to 5 sites was shown here. Delivery group, 50µL-IO: 50µL DNA without EP; 50µL- 1EP: 50µL DNA with 1 EP on the injection site; 200µL-IO: 200µL DNA without EP; 200µL-1EP: 200µL DNA and 1 EP; 200µL-4EP: 200µL DNA and 4 EPs; 50µLx4-IO: 4 injections with 50µL DNA without EP; 50µLx4-4EP: 4 injections with 50µL DNA and each EP on the injection site. B, Hematoxylin & eosin-stained skin samples. One representative of 3 treated sites was presented here for post delivery day 2 or day 7. Arrows are indicated the focal cell vacuolization. (magnification = 200, scale bar=100µm).
Figure 4
Figure 4. DNA distribution in the skin after i.d. DNA injection and non-invasive EP
A, B, C, Skin observation by flurorescence stereoscope after delivery with Fluorescein-labeled plasmid. Pictures were taken at post-delivery 1 hour. One representative picture of 2 sites was shown here. Delivery group: A, control; B, 50µL-IO: 50µL DNA without EP; C, 50µL-1EP: 50µL DNA with 1 EP on the injection site. D, E, total 4 cryosections (2 sections per sample) of each delivery were analyzed. Cell nuclei were blue-stained by DAPI. Cy™3-labeled DNA was shown red as indicated by arrows. D, One representative section of 50µL-IO was presented. E, One representative section of 50µL-EP was presented (magnification = 100, scale bar=100µm).
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References

    1. Heller R, Jaroszeski M, Atkin A, Moradpour D, Gilbert R, Wands J, Nicolau C. In vivo gene electroinjection and expression in rat liver. FEBS Lett. 1996;389(3):225–228. - PubMed
    1. Nishi T, Yoshizato K, Yamashiro S, Takeshima H, Sato K, Hamada K, Kitamura I, Yoshimura T, Saya H, Kuratsu J, Ushio Y. High-efficiency in vivo gene transfer using intraarterial plasmid DNA injection following in vivo electroporation. Cancer Res. 1996;56(5):1050–1055. - PubMed
    1. Sugimura K, Harimoto K, Kishimoto T. [In vivo gene transfer methods into bladder without viral vectors] Hinyokika Kiyo. 1997;43(11):823–827. - PubMed
    1. Rols MP, Delteil C, Golzio M, Dumond P, Cros S, Teissie J. In vivo electrically mediated protein and gene transfer in murine melanoma. Nat Biotechnol. 1998;16(2):168–171. - PubMed
    1. Mir LM. Nucleic acids electrotransfer-based gene therapy (electrogenetherapy): past, current, and future. Mol Biotechnol. 2009;43(2):167–176. - PubMed

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