Gentamicin induces functional type VII collagen in recessive dystrophic epidermolysis bullosa patients
David T. Woodley, … , Douglas Keene, Mei Chen
David T. Woodley, … , Douglas Keene, Mei Chen
Published July 10, 2017
Citation Information: J Clin Invest. 2017;127(8):3028-3038. https://doi.org/10.1172/JCI92707.
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Clinical Medicine Dermatology

Gentamicin induces functional type VII collagen in recessive dystrophic epidermolysis bullosa patients

Abstract

BACKGROUND. Recessive dystrophic epidermolysis bullosa (RDEB) is an incurable disease caused by mutations in the gene encoding type VII collagen, the major component of anchoring fibrils (AF). We previously demonstrated that gentamicin produced functional type VII collagen in RDEB cells harboring nonsense mutations. Herein, we determined whether topical or intradermal gentamicin administration induces type VII collagen and AFs in RDEB patients. METHODS. A double-blind, placebo-controlled pilot trial assessed safety and efficacy of topical and intradermal gentamicin in 5 RDEB patients with nonsense mutations. The topical arm tested 0.1% gentamicin ointment or placebo application 3 times daily at 2 open erosion sites for 2 weeks. The intradermal arm tested daily intradermal injection of gentamicin solution (8 mg) or placebo into 2 intact skin sites for 2 days in 4 of 5 patients. Primary outcomes were induction of type VII collagen and AFs at the test sites and safety assessment. A secondary outcome assessed wound closure of topically treated erosions. RESULTS. Both topical and intradermal gentamicin administration induced type VII collagen and AFs at the dermal-epidermal junction of treatment sites. Newly created type VII collagen varied from 20% to 165% of that expressed in normal human skin and persisted for 3 months. Topical gentamicin corrected dermal-epidermal separation, improved wound closure, and reduced blister formation. There were no untoward side effects from gentamicin treatments. Type VII collagen induction did not generate anti–type VII collagen autoantibodies in patients’ blood or skin. CONCLUSION. Topical and intradermal gentamicin suppresses nonsense mutations and induces type VII collagen and AFs in RDEB patients. Gentamicin therapy may provide a readily available treatment for RDEB patients with nonsense mutations. TRIAL REGISTRATION. ClinicalTrials.gov NCT02698735. FUNDING. Epidermolysis Bullosa Research Partnership, Epidermolysis Bullosa Medical Research Foundation, NIH, and VA Merit Award.

Authors

David T. Woodley, Jon Cogan, Yingping Hou, Chao Lyu, M. Peter Marinkovich, Douglas Keene, Mei Chen

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Figure 5

Topical and intradermal gentamicin do not induce anti–type VII collagen antibodies.

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Topical and intradermal gentamicin do not induce anti–type VII collagen ...
(A) Sera were obtained from RDEB patients before and 1 month after receiving gentamicin treatments and subjected to an ELISA using a commercially available ELISA kit against type VII collagen. Note that low-level anti–type VII collagen IgG antibodies were detected in 2 of the RDEB patients at day 0. However, none of the patients’ sera exhibited any increases in anti–type VII collagen IgG antibodies at 1 month after gentamicin treatment. Serum from a patient with EBA was used as a positive control, while serum from a normal human subject (NHS) was used as a negative control. Data represent the mean ± SD. (B) Skin biopsies from the gentamicin-treated test sites were obtained 1 month and 3 months after treatment and subjected to direct IF using an FITC-conjugated goat anti-human IgG antibody. Note that no IgG deposits of human anti–type VII collagen IgG were detected in any of the test sites. Scale bar: 50 μm.