Lentivirus gene therapy for purine nucleoside phosphorylase deficiency.

BACKGROUND: Purine nucleoside phosphorylase (PNP) deficiency causes the accumulation of toxic purine metabolites and lethal T cell immune defects, which might be corrected by expressing PNP by transplanting bone marrow (BM) cells transduced with lentiviral vectors containing the human PNP gene (lentiPNP).

METHODS: Lymphocytes from a single PNP-deficient patient as well as lymphocytes, fibroblasts and BM from PNP-deficient (PNP (-/-)) mice were transduced with lentiPNP. Female PNP (-/-) mice were transplanted with lentiPNP transduced BM cells from male PNP (-/-) mice or normal BM.

RESULTS: LentiPNP transduction significantly increased PNP expression in PNP-deficient human lymphocytes, murine lymphocytes, fibroblasts and BM cells. LentiPNP transduction also significantly improved the proliferation of PNP (-/-) murine lymphocyte and survival of irradiated PNP (-/-) fibroblasts. Polymerase chain reaction analysis demonstrated efficient transduction of lentiPNP into total and lineage-depleted BM cells grown ex vivo. LentiPNP transduced PNP (-/-) BM cells transplanted into PNP (-/-) mice expressed PNP in vivo, partially restored urinary uric acid secretion, improved thymocytes maturation, increased weight gain and extended survival of the mice. However, 12 weeks after transplant, the benefit of lentiPNP transduced cells and normal BM diminished and the percentage of engrafted donor cells decreased.

CONCLUSIONS: This short-term observational study provides the first in vivo proof that gene therapy may correct some of the abnormalities associated with PNP deficiency. Better gene transduction and expression, as well as improved cell engraftment, are required to further advance PNP gene therapy.