Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer

Ana Rovira, Maria De Angioletti, Olga Camacho-Vanegas, Delong Liu, Vittorio Rosti, Humilidad F. Gallardo, Rosario Notaro, Michel Sadelain, Lucio Luzzatto

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Many mutations of the housekeeping gene encoding glucose-6-phosphate dehydrogenase (G6PD) cause G6PD deficiency in humans. Some underlie severe forms of chronic nonspherocytic hemolytic anemia (CNSHA) for which there is no definitive treatment. By using retroviral vectors pseudotyped with the vesicular stomatitis virus G glycoprotein that harbor the human G6PD (hG6PD) complementary DNA, stable and lifelong expression of hG6PD was obtained in all the hematopoietic tissues of 16 primary bone marrow transplant (BMT) recipient mice and 14 secondary BMT recipients. These findings demonstrate the integration of a functional gene in totipotent stem cells. The average total G6PD in peripheral blood cells of these transplanted mice, measured as enzyme activity, was twice that of untransplanted control mice. This allowed the inference that the amount of G6PD produced by the transduced gene must be therapeutically effective. With the same vectors both the cloning efficiency and the ability to form embryoid bodies were restored in embryonic stem cells, in which the G6PD gene had been inactivated by targeted homologous recombination, thus effectively rescuing their defective phenotype. Finally, expression of normal human G6PD in hG6PD-deficient primary hematopoietic cells and in human hematopoietic cells engrafted in nonobese diabetic/severe combined immuno-deficient mice was obtained. This approach could cure severe CNSHA caused by G6PD deficiency.

Original languageEnglish
Pages (from-to)4111-4117
Number of pages7
JournalBlood
Volume96
Issue number13
Publication statusPublished - 2000

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Gene transfer
Glucosephosphate Dehydrogenase Deficiency
Glucosephosphate Dehydrogenase
Stem cells
Stem Cells
Genes
Hemolytic Anemia
Transplants
Totipotent Stem Cells
Bone Marrow
Embryoid Bodies
Genetic Vectors
Bone
Vesicular Stomatitis
Homologous Recombination
Essential Genes
Embryonic Stem Cells
Gene encoding
Blood Cells
Glycoproteins

ASJC Scopus subject areas

  • Hematology

Cite this

Rovira, A., De Angioletti, M., Camacho-Vanegas, O., Liu, D., Rosti, V., Gallardo, H. F., ... Luzzatto, L. (2000). Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer. Blood, 96(13), 4111-4117.

Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer. / Rovira, Ana; De Angioletti, Maria; Camacho-Vanegas, Olga; Liu, Delong; Rosti, Vittorio; Gallardo, Humilidad F.; Notaro, Rosario; Sadelain, Michel; Luzzatto, Lucio.

In: Blood, Vol. 96, No. 13, 2000, p. 4111-4117.

Research output: Contribution to journalArticle

Rovira, A, De Angioletti, M, Camacho-Vanegas, O, Liu, D, Rosti, V, Gallardo, HF, Notaro, R, Sadelain, M & Luzzatto, L 2000, 'Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer', Blood, vol. 96, no. 13, pp. 4111-4117.
Rovira A, De Angioletti M, Camacho-Vanegas O, Liu D, Rosti V, Gallardo HF et al. Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer. Blood. 2000;96(13):4111-4117.
Rovira, Ana ; De Angioletti, Maria ; Camacho-Vanegas, Olga ; Liu, Delong ; Rosti, Vittorio ; Gallardo, Humilidad F. ; Notaro, Rosario ; Sadelain, Michel ; Luzzatto, Lucio. / Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer. In: Blood. 2000 ; Vol. 96, No. 13. pp. 4111-4117.
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