Correction of purine nucleoside phosphorylase deficiency by retroviral-mediated gene transfer in mouse S49 T cell lymphoma: a model for gene therapy of T cell immunodeficiency.
To determine the effectiveness of retroviral-mediated purine nucleoside phosphorylase (PNP) gene transfer and expression for metabolic correction of PNP deficiency, we used as a gene transfer target the NSU-1 subline of murine S49 T lymphoma cells, an in vitro genetic model of PNP deficiency. NSU-1 cells were transduced with recombinant retroviruses that express either the murine or human PNP coding sequences under transcriptional regulation of the Moloney murine leukemia virus (Mo-MLV) long terminal repeat (LTR), resulting in expression of substantial levels of PNP activity. Untransduced or control virus-transduced NSU-1 cells were extremely sensitive to deoxyguanosine, a PNP substrate that is toxic for lymphoid cells. However, PNP-virus transduction of NSU-1 cells metabolically corrected the sensitivity of these cells to deoxyguanosine, resulting in near wild-type levels of growth inhibition. These results demonstrate that retroviral-mediated PNP gene transfer and expression corrects the metabolic defect observed in PNP-deficient murine lymphoid cells, suggesting that PNP gene transfer and expression in human lymphoid cells might similarly correct substrate-mediated toxicity and provide an effective genetic therapy.
Foresman
M D
MD
Institute of Human Genetics, University of Minnesota, Minneapolis 55455.
Nelson
D M
DM
McIvor
R S
RS
D000818:Animals; D003849:Deoxyguanosine / Q000494:pharmacology; D004351:Drug Resistance; D004790:Enzyme Induction; D015316:Genetic Therapy; D005822:Genetic Vectors; D007153:Immunologic Deficiency Syndromes / Q000235:genetics / Q000628:therapy*; D016399:Lymphoma, T-Cell / Q000473:pathology*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000235:genetics; D011401:Promoter Regions, Genetic; D011683:Purine-Nucleoside Phosphorylase / Q000172:deficiency*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000201:enzymology*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000201:enzymology
D011993:Recombinant Fusion Proteins; D011683:Purine-Nucleoside Phosphorylase; D003849:Deoxyguanosine