Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin

Marc G. Jeschke, Robert E. Barrow, Hal K. Hawkins, Ziming Tao, J. Regino Perez-Polo, David Herndon

Research output: Contribution to journalArticle

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Abstract

Gene therapy using cationic liposomes containing cDNA is a relatively new approach with great potential; however, little is known about the mechanisms of dermal gene transfer, its biodistribution, systemic transfection, and cellular uptake. This study identifies mechanisms, transfection rates, and biodistribution of liposomal gene transfers in the skin of thermally injured rats using cDNA gene constructs coding for insulin- like growth factor-I (IGF-I) and Lac Z. Male Sprague-Dawley rats (350 to 375 g) were given a 60% total body surface area full-thickness scald burn that was followed by weekly subcutaneous injections of normal saline (control, n = 10), liposomes plus 0.2 μg Lac Z cDNA construct driven by a cytomegalovirus (CMV) promoter (vehicle, n = 10), or liposomes containing 2.2 μg cDNA coding for IGF-I plus 0.2 μg Lac Z cDNA construct driven by a CMV promoter (IGF-I cDNA, n = 10). Gene transfection was determined by histochemical and luminescent β-galactosidase assays of blood, skin, liver, spleen, and kidney. Transcription of IGF-I cDNA to IGF-I mRNA was determined in skin cells by Northern blot analyses. Levels of IGF-I protein in blood, skin, liver, spleen, and kidney were measured by radioimmunoassay. The biological activity of the translated IGF-I was evaluated by the mitogenic activity in dermal cells and the rate of re-epithelization. Gene transfection was observed only in skin cells. The expression of IGF-I mRNA increased in skin cells of burned rats receiving liposomes containing the IGF-I cDNA construct compared with liposomes without the construct or normal saline. IGF-I protein levels in the skin of rats receiving the IGF-I cDNA was 176 ± 4 ng/ml compared with 105 ± 6 ng/ml for liposomes alone or 90 ± 3 ng/ml for saline (p < 0.05). The translated IGF-I protein was found biologically active in the skin by increasing skin cell proliferation and accelerating re-epithelization 33 days after thermal injury (p < 0.05). No systemic transfection could be detected. Skin cells transfected with liposomes encapsulating the IGF-I cDNA constructs increased the expression of IGF-I mRNA transcript and the expression of a biologically active IGF-I protein. Liposomes containing the cDNA coding for IGF-I present an effective approach to gene therapy in the skin.

Original languageEnglish (US)
Pages (from-to)151-158
Number of pages8
JournalLaboratory Investigation
Volume80
Issue number2
StatePublished - Feb 2000

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Insulin-Like Growth Factor I
Skin
Complementary DNA
Liposomes
Genes
Transfection
Cytomegalovirus
Genetic Therapy
Messenger RNA
Spleen
Galactosidases
Luminescent Measurements
Kidney
Proteins
Body Surface Area
Liver
Subcutaneous Injections
Northern Blotting
Radioimmunoassay
Sprague Dawley Rats

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Jeschke, M. G., Barrow, R. E., Hawkins, H. K., Tao, Z., Perez-Polo, J. R., & Herndon, D. (2000). Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin. Laboratory Investigation, 80(2), 151-158.

Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin. / Jeschke, Marc G.; Barrow, Robert E.; Hawkins, Hal K.; Tao, Ziming; Perez-Polo, J. Regino; Herndon, David.

In: Laboratory Investigation, Vol. 80, No. 2, 02.2000, p. 151-158.

Research output: Contribution to journalArticle

Jeschke, MG, Barrow, RE, Hawkins, HK, Tao, Z, Perez-Polo, JR & Herndon, D 2000, 'Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin', Laboratory Investigation, vol. 80, no. 2, pp. 151-158.
Jeschke MG, Barrow RE, Hawkins HK, Tao Z, Perez-Polo JR, Herndon D. Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin. Laboratory Investigation. 2000 Feb;80(2):151-158.
Jeschke, Marc G. ; Barrow, Robert E. ; Hawkins, Hal K. ; Tao, Ziming ; Perez-Polo, J. Regino ; Herndon, David. / Biodistribution and feasibility of non-viral IGF-I gene transfers in thermally injured skin. In: Laboratory Investigation. 2000 ; Vol. 80, No. 2. pp. 151-158.
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