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

Authors: Rovira, A.; De Angioletti, M.; Camacho Vanegas, O.; Liu, D.; Rosti, V.; Gallardo, H. F.; Notaro, R.; Sadelain, M.; Luzzatto, L.
Article Title: Stable in vivo expression of glucose-6-phosphate dehydrogenase (G6PD) and rescue of G6PD deficiency in stem cells by gene transfer
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. © 2000 by The American Society of Hematology.
Keywords: genetics; mouse; phenotype; animal; cytology; metabolism; mouse mutant; animals; mice; bone marrow cells; genetic transcription; transcription, genetic; enzymology; pathology; mice, scid; transfection; mice, inbred c57bl; c57bl mouse; gene vector; genetic vectors; stem cell; genetic transfection; xenograft; virus rna; membrane glycoproteins; membrane protein; transgene; hematopoietic stem cells; transplantation, heterologous; stem cells; bone marrow cell; hematopoietic stem cell; graft survival; mice, inbred nod; bone marrow transplantation; nonobese diabetic mouse; isolation and purification; moloney leukemia oncovirus; moloney murine leukemia virus; cell strain 3t3; transgenes; complementary dna; rna, viral; 3t3 cells; enzyme induction; dna, complementary; genetic complementation; genetic complementation test; vesicular stomatitis virus; glucose 6 phosphate dehydrogenase deficiency; glucose 6 phosphate dehydrogenase; glucosephosphate dehydrogenase; viral envelope proteins; virus envelope protein; humans; human; male; article; radiation chimera; vesicular stomatitis-indiana virus; provirus; proviruses; g protein, vesicular stomatitis virus; genetic reassortment; glucosephosphate dehydrogenase deficiency; reassortant viruses
Journal Title: Blood
Volume: 96
Issue: 13
ISSN: 0006-4971
Publisher: American Society of Hematology  
Date Published: 2000-12-15
Start Page: 4111
End Page: 4117
Language: English
PUBMED: 11110681
PROVIDER: scopus
Notes: Export Date: 18 November 2015 -- Source: Scopus