Therapeutic success and efficacy of nonviral liposomal cDNA gene transfer to the skin in vivo is dose dependent

Jeschke, Marc G.; Richter, Günther; Herndon, David N.; Geissler, Edward K.; Hartl, Michael; Hofstätter, F.; Jauch, Karl Walter; Perez‐Polo, J. Regino

doi:10.1038/sj.gt.3301589

Cited by 28 publications

(15 citation statements)

References 44 publications

(80 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…There are several possible candidate genes. The majority of these may be involved in wound healing, such as keratinocyte growth factor [59], epidermal growth factor, or members of the fibroblast growth factor family [60]. Genes involved in immune regulation may also be investigated, after increasing evidence of an immune aetiology for PG [3,61,62,63,64,65,66].…”

Section: Discussionmentioning

confidence: 99%

Gene Therapy for Pyoderma Gangrenosum: Optimal Transfection Conditions and Effect of Drugs on Gene Delivery in the HaCaT Cell Line Using Cationic Liposomes

Teagle¹,

Birchall

Hargest³

2016

Skin Pharmacol Physiol

View full text Add to dashboard Cite

Background/Aims: Pyoderma gangrenosum (PG) is a rare ulcerative skin disease, currently treated empirically with immunosuppression. PG is a good target for gene therapy since the skin is easily accessible. This study used the FDA-approved vector Lipofectamine® 2000 to investigate in vitro transfection of skin keratinocytes. The aim was to determine an optimum transfection protocol, including the effect of drugs currently used to treat PG on the efficiency of gene transfer, since gene therapy is unlikely to be used as monotherapy. Methods: Cells of the HaCaT line were transfected with the lacZ reporter gene, and transgene expression was measured after a given time period. Conditions tested were: relative concentrations of DNA and Lipofectamine®, time from transfection to measurement of expression, pH, and exposure to clinically relevant drugs (hydrocortisone, methotrexate, infliximab). Results: The greatest levels of β-galactosidase expression were observed using a DNA:Lipofectamine® ratio of 1:5 (μg/μl) on day 3 after transfection, using culture medium at pH 7, and in the presence of hydrocortisone. Transfection efficiency was reduced by the presence of methotrexate and not significantly affected by infliximab. Conclusion: Gene therapy is a potential future strategy for the management of PG; this study is a step towards the development of a topical gene-based agent.

show abstract

Section: Discussionmentioning

confidence: 99%

Gene Therapy for Pyoderma Gangrenosum: Optimal Transfection Conditions and Effect of Drugs on Gene Delivery in the HaCaT Cell Line Using Cationic Liposomes

Teagle¹,

Birchall

Hargest³

2016

Skin Pharmacol Physiol

View full text Add to dashboard Cite

show abstract

“…In a study using two plasmids coding for keratinocyte growth factor and insulin-like growth factor, three different doses of 0.22 µg, 2.2 µg and 22 µg DNA were injected into the skin. Therapeutic parameters characteristic for wound healing were monitored, and the highest therapeutic effect for both plasmids was shown for the 2.2 µg doses [103]. This type of dose dependency was not shown for topically applied DNA yet.…”

Section: Formulations and Delivery Systemsmentioning

confidence: 91%

DNA Delivery for Vaccination and Therapeutics Through the Skin

Földvári¹,

Babiuk²,

Badea

2006

CDD

View full text Add to dashboard Cite

Cutaneous gene therapy and DNA vaccination are potential applications of plasmid delivery methods where a gene for an antigen or a therapeutic protein is inserted in the plasmid and applied to the skin. However, the delivery of the DNA plasmid is a major challenge due to the unusual physicochemical properties of the DNA, the tissue and cellular barriers and expression difficulties. Even though the skin is the most accessible organ of the body and it is an ideal target for gene therapy, the delivery of plasmid DNA across the skin is very difficult due to the specific barrier function of the stratum corneum and the inconsistent transfection rate of keratinocytes and other epidermal cells. To date there is no gene delivery system that was shown to be optimal for cutaneous gene therapy. In order to develop an efficient non-viral delivery vehicle we need to design a system that provides the combined properties of effective DNA condensation, cutaneous permeation, cellular transfection and sufficiently sustained expression. This paper reviews the formulation approaches and delivery methods for DNA through the skin in the context of the barriers both at the tissue and cellular levels for both vaccine and gene therapy applications.

show abstract

“…This reagent interacts spontaneously with IGF-I cDNA to form the lipid cDNA complex. The dose of 2.2 g for the IGF-I cDNA was defined in a dose-response study (11).…”

Section: Methodsmentioning

confidence: 99%

Exogenous liposomal IGF-I cDNA gene transfer leads to endogenous cellular and physiological responses in an acute wound

Jeschke¹,

Schubert²,

Klein³

2004

American Journal of Physiology-Regulatory, Integrative and Comp

View full text Add to dashboard Cite

The purpose of the present study was to examine whether exogenous liposomal cDNA gene transfer is recognized by the cell and causes endogenous cellular and physiological responses. When administered as a protein, IGF-I is known to cause adverse side effects due to lack of cellular responses. Therefore, we used IGF-I cDNA as a vector to study cellular and physiological effects after liposomal administration to wounded skin. Sprague-Dawley rats were given a scald burn to inflict an acute wound and were divided into two groups to receive weekly subcutaneous injections of liposomes plus the Lac-Z gene (0.2 μg vehicle) or liposomes plus the IGF-I cDNA (2.2 μg) and Lac Z gene (0.22 μg). Transfection was confirmed by histochemical assays for β-galactosidase. Planimetry, immunological assays, and histological and immunohistochemical techniques were used to determine molecular mechanisms after gene transfer, protein expression, and dermal and epidermal regeneration. IGF-I cDNA transfer increased IGF-I protein expression and caused concomitant cellular responses by increasing IGF binding protein (IGFBP)-3 and decreasing IGFBP-1. IGF-I cDNA gene transfer increased keratinocyte growth factor expression and exerted promitogenic antiapoptotic effects on basal keratinocytes, thus improving epidermal regeneration. IGF-I cDNA improved dermal regeneration by an increased collagen deposition and morphology. IGF-I cDNA increased VEGF concentrations and thus neovascularization. Exogenous-administered IGF-I cDNA is recognized by the cell and leads to similar intracellular responses as the endogenous gene. Liposomal IGF-I gene transfer further leads to improved dermal and epidermal regeneration by interacting with other growth factors.

show abstract

Therapeutic success and efficacy of nonviral liposomal cDNA gene transfer to the skin in vivo is dose dependent

Abstract: It is well documented that responses to growth factor treat-

Cited by 28 publications

References 44 publications

Gene Therapy for Pyoderma Gangrenosum: Optimal Transfection Conditions and Effect of Drugs on Gene Delivery in the HaCaT Cell Line Using Cationic Liposomes

Gene Therapy for Pyoderma Gangrenosum: Optimal Transfection Conditions and Effect of Drugs on Gene Delivery in the HaCaT Cell Line Using Cationic Liposomes

DNA Delivery for Vaccination and Therapeutics Through the Skin

Exogenous liposomal IGF-I cDNA gene transfer leads to endogenous cellular and physiological responses in an acute wound

Contact Info

Product

Resources

About