Halting angiogenesis by non-viral somatic gene therapy alleviates psoriasis and murine psoriasiform skin lesions
John R. Zibert, … , Lone Skov, Michael P. Schön
John R. Zibert, … , Lone Skov, Michael P. Schön
Published January 4, 2011; First published December 6, 2010
Citation Information: J Clin Invest. 2011;121(1):410-421. https://doi.org/10.1172/JCI41295.
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Research Article Dermatology

Halting angiogenesis by non-viral somatic gene therapy alleviates psoriasis and murine psoriasiform skin lesions

Abstract

Dysregulated angiogenesis is a hallmark of chronic inflammatory diseases, including psoriasis, a common skin disorder that affects approximately 2% of the population. Studying both human psoriasis in 2 complementary xenotransplantation models and psoriasis-like skin lesions in transgenic mice with epidermal expression of human TGF-β1, we have demonstrated that antiangiogenic non-viral somatic gene therapy reduces the cutaneous microvasculature and alleviates chronic inflammatory skin disorders. Transient muscular expression of the recombinant disintegrin domain (RDD) of metargidin (also known as ADAM-15) by in vivo electroporation reduced cutaneous angiogenesis and vascularization in all 3 models. As demonstrated using red fluorescent protein–coupled RDD, the treatment resulted in muscular expression of the gene product and its deposition within the cutaneous hyperangiogenic connective tissue. High-resolution ultrasound revealed reduced cutaneous blood flow in vivo after electroporation with RDD but not with control plasmids. In addition, angiogenesis- and inflammation-related molecular markers, keratinocyte proliferation, epidermal thickness, and clinical disease scores were downregulated in all models. Thus, non-viral antiangiogenic gene therapy can alleviate psoriasis and may do so in other angiogenesis-related inflammatory skin disorders.

Authors

John R. Zibert, Katrin Wallbrecht, Margarete Schön, Lluis M. Mir, Grete K. Jacobsen, Veronique Trochon-Joseph, Céline Bouquet, Louise S. Villadsen, Ruggero Cadossi, Lone Skov, Michael P. Schön

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Figure 1

Expression and functional activity of RDD fusion proteins in vitro.

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Expression and functional activity of RDD fusion proteins in vitro. (A) ...
(A) Human fibroblasts transfected with different constructs were analyzed for RDD expression by RT-PCR (lanes: 1, weight marker; 2, untransfected fibroblasts; 3, transfected with pVAX-mock; 4, transfected with pVAX-RDD; 5, transfected with pVAX-RFP-RDD; 6, negative control; 7, plasmid pVAX-RDD [positive control 1]; 8, plasmid pVAX-RFP-RDD [positive control 2]. (B) Murine LL/2 cells were transfected with the pVAX-RFP-RDD or the pVAX-RFP plasmid as indicated. Protein expression was detected by fluorescence microscopy. Original magnification, ×400. (C) HUVEC cultures were scratched in a standardized fashion (wounding of the monolayer), thus creating a cell-free lane (width, 0.5 mm) in each culture. Cultures were then incubated with supernatant from untransfected human fibroblasts (circles), from human fibroblasts transfected with the pVAX-mock plasmid (squares), or the pVAX-RDD plasmid (triangles). Migration of cells into the cell-free lane was monitored. Each data point represents the mean of 10 measurements (± SD). Units on the y-axis: μm. ***P < 0.001. (D) Representative HUVEC cultures described in C. The controls shown here are cultures incubated with supernatant from pVAX-mock–transfected fibroblasts, which were similar to the other controls. The margins of the cell layers are indicated by black lines. Original magnification, ×100. (E) Expression of the RDD target proteins, integrins αVβ3 and α5β1, on HUVEC as detected by flow cytometry. Strong signals are detected with antibodies directed against CD49e (α5 integrin subunit), CD51 (αV integrin subunit), and a combinatorial epitope of CD51 and CD61 (the αVβ3 integrin heterodimer).