Molecular basis for the dystrophic forms of epidermolysis bullosa: mutations in the type VII collagen gene

Uitto, Jouni; Christiano, Angela M.

doi:10.1007/bf00370713

Cited by 111 publications

(50 citation statements)

References 34 publications

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“…The pathogenesis of DEB involves a basement membrane zone (BMZ)-specific collagen called type VII collagen, the major component of structures within the BMZ called anchoring fibrils. DEB patients have defects in the gene, designated COL7A1, that encodes for type VII (anchoring fibril) collagen (2). The BMZ of patients with DEB is characterized by a paucity or diminutive size of anchoring fibrils (3).…”

Section: Dystrophic Epidermolysis Bullosa (Deb)mentioning

confidence: 99%

See 1 more Smart Citation

Development and Characterization of A Recombinant Truncated Type VII Collagen “Minigene”

Chen¹,

O’Toole²,

Muellenhoff

et al. 2000

Journal of Biological Chemistry

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Dystrophic epidermolysis bullosa (DEB) is an inherited mechano-bullous disorder of skin caused by mutations in the type VII collagen gene. The lack of therapy for DEB provides an impetus to develop gene therapy strategies. However, the full-length 9-kilobase type VII collagen cDNA exceeds the cloning capacity of current viral delivery vectors. In this study, we produced a recombinant type VII minicollagen containing the intact noncollagenous domains, NC1 and NC2, and part of the central collagenous domain using stably transfected human 293 cell clones and purified large quantities of the recombinant minicollagen VII from culture media. Minicollagen VII was secreted as correctly-folded, disulfidebonded, helical trimers resistant to protease degradation. Purified minicollagen VII bound to fibronectin, laminin-5, type I collagen, and type IV collagen. Furthermore, retroviral-mediated transduction of the minigene construct into DEB keratinocytes (in which type VII collagen was absent) resulted in persistent synthesis and secretion of a 230-kDa recombinant minicollagen VII. In comparison with parent DEB keratinocytes, the gene-corrected DEB keratinocytes demonstrated enhanced cell-substratum adhesion, increased proliferative potential, and reduced cell motility, features that reversed the DEB phenotype toward normal. We conclude that the use of the minicollagen VII may provide a strategy to correct the cellular manifestations of gene defects in DEB. Dystrophic epidermolysis bullosa (DEB)1 is a group of heritable mechano-bullous skin diseases characterized by skin fragility, separation of the epidermis from the dermis (blister formation), milia and scarring (1). The pathogenesis of DEB involves a basement membrane zone (BMZ)-specific collagen called type VII collagen, the major component of structures within the BMZ called anchoring fibrils. DEB patients have defects in the gene, designated COL7A1, that encodes for type VII (anchoring fibril) collagen (2). The BMZ of patients with DEB is characterized by a paucity or diminutive size of anchoring fibrils (3). Earlier genetic linkage studies identified the human COL7A1 as the gene responsible for DEB (2, 4 -6), and then numerous COL7A1 gene defects were directly identified in families of patients with DEB (2).Type VII collagen is composed of three identical ␣ chains, each consisting of a 145-kDa central collagenous triple-helical segment characterized by repeating Gly-X-Y amino acid sequences, flanked by a large 145-kDa amino-terminal, noncollagenous domain (NC1), and a small 34-kDa carboxyl-terminal non-collagenous domain (NC2) (7,8). Within the extracellular space, type VII collagen molecules form antiparallel, tail-to-tail dimers stabilized by disulfide bonding through a small carboxyl-terminal NC2 overlap between two type VII collagen molecules. A portion of the NC2 domain is then proteolytically removed (9). The antiparallel dimers then aggregate laterally to form anchoring fibrils with large globular NC1 domains at both ends of the structure. NC1 domains have been...

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Section: Dystrophic Epidermolysis Bullosa (Deb)mentioning

confidence: 99%

“…The BMZ of patients with DEB is characterized by a paucity or diminutive size of anchoring fibrils (3). Earlier genetic linkage studies identified the human COL7A1 as the gene responsible for DEB (2, 4 -6), and then numerous COL7A1 gene defects were directly identified in families of patients with DEB (2).…”

Section: Dystrophic Epidermolysis Bullosa (Deb)mentioning

confidence: 99%

Development and Characterization of A Recombinant Truncated Type VII Collagen “Minigene”

Chen¹,

O’Toole²,

Muellenhoff

et al. 2000

Journal of Biological Chemistry

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“…RDEB is caused by mutations in the COL7A1 gene that encodes type VII collagen, the major component of anchoring fibrils (AFs) that tether together the 2 main layers of skin, the epidermis and dermis (2,3). With insufficient type VII collagen and AFs, the adherence between the epidermis and dermis is compromised and subepidermal blistering from minor trauma ensues.…”

Section: Introductionmentioning

confidence: 99%

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

Woodley¹,

Cogan²,

Hou³

et al. 2017

Journal of Clinical Investigation

100

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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.

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“…14 Mutations or deficiencies of collagen type VII as in bullous systemic lupus erythematosus or dystrophic forms of EB 15 result in a paucity of collagen type VII anchoring collagen fibrils 16 leading to skin fragility and extensive blistering. Patients with skin basement membrane disorders constantly suffer from severe blisters and recurring wounds throughout the body and associated infections.…”

Section: Introductionmentioning

confidence: 99%

Superficial Dermal Fibroblasts Enhance Basement Membrane and Epidermal Barrier Formation in Tissue-Engineered Skin: Implications for Treatment of Skin Basement Membrane Disorders

Varkey

Ding

Tredget

2013

Tissue Engineering Part A

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Basement membrane is a highly specialized structure that binds the dermis and the epidermis of the skin, and is mainly composed of laminins, nidogen, collagen types IV and VII, and the proteoglycans, collagen type XVIII and perlecan, all of which play critical roles in the function and resilience of skin. Both dermal fibroblasts and epidermal keratinocytes contribute to the development of the basement membrane, and in turn the basement membrane and underlying dermis influence the development and function of the epidermal barrier. Disruption of the basement membrane results in skin fragility, extensive painful blistering, and severe recurring wounds as seen in skin basement membrane disorders such as epidermolysis bullosa, a family of life-threatening congenital skin disorders. Currently, there are no successful strategies for treatment of these disorders; we propose the use of tissue-engineered skin as a promising approach for effective wound coverage and to enhance healing. Fibroblasts and keratinocytes isolated from superficial and deep dermis and epidermis, respectively, of tissue from abdominoplasty patients were independently cocultured on collagen-glycosaminoglycan matrices, and the resulting tissue-engineered skin was assessed for functional differences based on the underlying specific dermal fibroblast subpopulation. Tissue-engineered skin with superficial fibroblasts and keratinocytes formed a continuous epidermis with increased epidermal barrier function and expressed higher levels of epidermal proteins, keratin-5, and E-cadherin, compared to that with deep fibroblasts and keratinocytes, which had an intermittent epidermis. Further, tissue-engineered skin with superficial fibroblasts and keratinocytes formed better basement membrane, and produced more laminin-5, nidogen, collagen type VII, compared to that with deep fibroblasts and keratinocytes. Overall, our results demonstrate that tissue-engineered skin with superficial fibroblasts and keratinocytes forms significantly better basement membrane with higher expression of dermo-epidermal adhesive and anchoring proteins, and superior epidermis with enhanced barrier function compared to that with deep fibroblasts and keratinocytes, or with superficial fibroblasts, deep fibroblasts, and keratinocytes. The specific use of superficial fibroblasts in tissue-engineered skin may thus be more beneficial to promote adhesion of newly formed skin and wound healing, and is therefore promising for the treatment of patients with basement membrane disorders and other skin blistering diseases.

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Molecular basis for the dystrophic forms of epidermolysis bullosa: mutations in the type VII collagen gene

Cited by 111 publications

References 34 publications

Development and Characterization of A Recombinant Truncated Type VII Collagen “Minigene”

Development and Characterization of A Recombinant Truncated Type VII Collagen “Minigene”

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

Superficial Dermal Fibroblasts Enhance Basement Membrane and Epidermal Barrier Formation in Tissue-Engineered Skin: Implications for Treatment of Skin Basement Membrane Disorders

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