Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue
Susana Ortiz-Urda, … , M. Peter Marinkovich, Paul A. Khavari
Susana Ortiz-Urda, … , M. Peter Marinkovich, Paul A. Khavari
Published January 15, 2003
Citation Information: J Clin Invest. 2003;111(2):251-255. https://doi.org/10.1172/JCI17193.
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Article Dermatology

Injection of genetically engineered fibroblasts corrects regenerated human epidermolysis bullosa skin tissue

Abstract

Current therapeutic strategies for genetic skin disorders rely on the complex process of grafting genetically engineered tissue to recipient wound beds. Because fibroblasts synthesize and secrete extracellular matrix, we explored their utility in recessive dystrophic epidermolysis bullosa (RDEB), a blistering disease due to defective extracellular type VII collagen. Intradermal injection of RDEB fibroblasts overexpressing type VII collagen into intact RDEB skin stably restored correctly localized type VII collagen expression in vivo and normalized hallmark RDEB disease features, including subepidermal blistering and anchoring fibril defects.

Authors

Susana Ortiz-Urda, Qun Lin, Cheryl L. Green, Douglas R. Keene, M. Peter Marinkovich, Paul A. Khavari

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Figure 1

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Type VII collagen expression in primary skin cells. (a) Immunoblots of e...
Type VII collagen expression in primary skin cells. (a) Immunoblots of extracts from primary RDEB (EB) and normal (NL) patient skin fibroblasts and keratinocytes (KCs). The first lane represents RDEB+ fibroblasts engineered to overexpress type VII collagen. pCOL7A1 is a CMV-driven expression plasmid for type VII collagen (3) stably transfected using the φC31 integrase. Optical densitometric quantitation of type VII collagen protein levels are noted below each sample lane, normalized to BRG1 (25), a constitutively expressed control for extract loading, quality, and transfer; normal fibroblasts are assigned a relative value of 1.0. UD, undetectable. (b) Cellular expression of type VII protein expression (green) in engineered (RDEB+), normal (NL), and uncorrected (RDEB–) fibroblasts. Counterstaining with Hoechst 33342 marks all cellular nuclei (blue).