Adenoviral gene transfer restores lysyl hydroxylase activity in type VI Ehlers-Danlos syndrome.
Type VI Ehlers-Danlos syndrome is a disease characterized by disturbed lysine hydroxylation of collagen. The disease is caused by mutations in lysyl hydroxylase 1 gene and it affects several organs including the cardiovascular system, the joint and musculoskeletal system, and the skin. The skin of type VI Ehlers-Danlos syndrome patients is hyperelastic, scars easily, and heals slowly and poorly. We hypothesized that providing functional lysyl hydroxylase 1 gene to the fibroblasts in and around wounds in these patients would improve healing. In this study we tested the feasibility of transfer of the lysyl hydroxylase 1 gene into fibroblasts derived from rats and a type VI Ehlers-Danlos syndrome patient (in vitro) and into rat skin (in vivo). We first cloned human lysyl hydroxylase 1 cDNA into a recombinant adenoviral vector (Ad5RSV-LH). Transfection of human type VI Ehlers-Danlos syndrome fibroblasts (about 20% of normal lysyl hydroxylase 1 activity) with the vector increased lysyl hydroxylase 1 activity in these cells to near or greater levels than that of wild type, unaffected fibroblasts. The adenoviral vector successfully transfected rat fibroblasts producing both beta-galactosidase and lysyl hydroxylase 1 gene activity. We next expanded our studies to a rodent model. Intradermal injections of the vector to the abdominal skin of rats produced lysyl hydroxylase 1 mRNA and elevated lysyl hydroxylase 1 activity, in vivo. These data suggest the feasibility of gene replacement therapy to modify skin wound healing in type VI Ehlers-Danlos syndrome patients.
Rauma
T
T
Department of Pharmacology and Toxicology, University of Oulu, Finland.
Kumpumäki
S
S
Anderson
R
R
Davidson
B L
BL
Ruotsalainen
H
H
Myllylä
R
R
Hautala
T
T
D000256:Adenoviridae / Q000235:genetics*; D004535:Ehlers-Danlos Syndrome / Q000145:classification* / Q000201:enzymology* / Q000473:pathology; D005347:Fibroblasts / Q000201:enzymology; D005696:Galactosidases / Q000235:genetics; D018014:Gene Transfer Techniques; D006801:Humans; D006901:Hydroxylysine / Q000378:metabolism; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012867:Skin / Q000201:enzymology / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology
D012333:RNA, Messenger; D006901:Hydroxylysine; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase; D005696:Galactosidases