BP180 gene delivery in junctional epidermolysis bullosa.
Epidermolysis bullosa (EB) comprises a family of inherited blistering skin diseases for which current therapy is only palliative. Junctional EB (JEB) involves dissociation of the dermal-epidermal junction and results from mutations in a number of genes that encode vital structural proteins, including BP180 (type XVII collagen/BPAG2). In order to develop a model of corrective gene delivery for JEB, we produced a retroviral expression vector for wild-type human BP180 and used it to restore BP180 protein expression to primary keratinocytes from BP180-negative patients with generalized atrophic JEB. Restoration of full-length BP180 protein expression was associated with adhesion parameter normalization of primary JEB keratinocytes in vitro. These cells were then used to regenerate human skin on immune-deficient mice. BP180 gene-transduced tissue demonstrated restoration of BP180 gene expression at the dermal-epidermal junction in vivo while untransduced regenerated JEB skin entirely lacked BP180 expression. These findings provide a basis for future efforts to achieve gene delivery in human EB skin tissue.
Seitz
C S
CS
VA Palo Alto Health Care System, CA 94304, USA.
Giudice
G J
GJ
Balding
S D
SD
Marinkovich
M P
MP
Khavari
P A
PA
D000818:Animals; D001324:Autoantigens / Q000235:genetics* / Q000378:metabolism; D002352:Carrier Proteins; D002448:Cell Adhesion; D002478:Cells, Cultured; D003094:Collagen; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D016109:Epidermolysis Bullosa, Junctional / Q000628:therapy*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D015603:Keratinocytes / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D012190:Retroviridae / Q000235:genetics; D012867:Skin / Q000378:metabolism; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D003094:Collagen; D003598:Cytoskeletal Proteins; D000071821:Dystonin; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D000097805:Collagen Type XVII; C486268:DST protein, human; C095667:Dst protein, mouse