Retrovirus-mediated gene transfer of ornithine-delta-aminotransferase into keratinocytes from gyrate atrophy patients.
Gyrate atrophy is a progressive blindness associated with deficiency of ornithine aminotransferase (OAT). The strategy of using an autologous keratinocyte graft, modified to express high levels of OAT as an ornithine-catabolizing skin-based enzyme sink, is investigated. Two OAT-containing retroviral vectors were constructed with or without a resistance gene. When packaged in a retroviral vector particle generated with the gibbon ape leukemia (GALV) virus envelope (PG13), these vectors could readily transduce >50% of target keratinocytes. The transduced keratinocytes in culture expressed up to 75-fold more OAT than normal control keratinocytes and these gene-modified cells extracted [14C]ornithine more efficiently than controls. The vector prepared without neo transduced cells more efficiently and led to higher levels of OAT expression than the neo-containing vector. Ornithine catabolism was maintained at high levels when the transduced patient keratinocytes were differentiated in vitro as a multilayered cutaneous organoid.
Jensen
T G
TG
Clinical Gene Therapy Branch, National Center for Human Genome Research, National Institutes of Health, Bethesda, MD 20892-1851, USA.
Sullivan
D M
DM
Morgan
R A
RA
Taichman
L B
LB
Nussenblatt
R B
RB
Blaese
R M
RM
Csaky
K G
KG
D016475:3T3 Cells; D000818:Animals; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D015799:Gyrate Atrophy / Q000201:enzymology* / Q000628:therapy; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics*; D051379:Mice; D009953:Ornithine-Oxo-Acid Transaminase / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
D009953:Ornithine-Oxo-Acid Transaminase