Retrovirus-mediated transfer of human alpha-galactosidase A gene to human CD34+ hematopoietic progenitor cells.

Fabry disease, caused by a deficiency of lysosomal enzyme alpha-galactosidase A (alpha-gal A), is one of the inherited disorders potentially treatable by gene transfer to hematopoietic stem cells. In this study, a high-titer amphotropic retroviral producer cell line, MFG-alpha-gal A, was established. CD34+ cells from normal umbilical cord blood were transduced by centrifugal enhancement. The alpha-gal A activity in transduced cells increased 3.6-fold above the activity in nontransduced cells. Transduction efficiency measured by PCR for the integrated alpha-gal A cDNA in CFU-GM colonies was in the range of 42-88% (average, 63%). The expression of functional enzyme in TFI erythroleukemia was sustained for as long as cells remained in culture (84 days) and for 28 days in LTC-IC cultures of CD34+ cells. The ability of the transduced CD34+ cells to secrete the enzyme and to correct enzyme-deficient Fabry fibroblasts was assessed by cocultivation of these cells. The enzyme was secreted into the medium from transduced CD34+ cells and taken up by Fabry fibroblasts through mannose 6-phosphate receptors. These findings suggest that genetically corrected hematopoietic stem/progenitor cells can be an enzymatic source for neighboring enzyme-deficient cells, and can potentially be useful for gene therapy of Fabry disease.

[1]  W. Swaney,et al.  Comparison of methods for retroviral mediated transfer of glucocerebrosidase gene to CD34+ hematopoietic progenitor cells , 1998, European journal of haematology.

[2]  K. Macdermot,et al.  Gene transfer and expression of humanα-galactosidase from mouse muscle in vitro andin vivo , 1997, Gene Therapy.

[3]  R. Brady,et al.  Correction in trans for Fabry disease: expression, secretion and uptake of alpha-galactosidase A in patient-derived cells driven by a high-titer recombinant retroviral vector. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[4]  T. Dexter,et al.  Long-term in vitro correction of alpha-L-iduronidase deficiency (Hurler syndrome) in human bone marrow. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[5]  A. Bahnson,et al.  Efficient retroviral mediated transfer of the glucocerebrosidase gene in CD34+ enriched umbilical cord blood human hematopoietic progenitors. , 1995, Experimental hematology.

[6]  A. Bahnson,et al.  Cytokine mobilization of peripheral blood stem cells in patients with Gaucher disease with a view to gene therapy. , 1995, Experimental hematology.

[7]  A. Bahnson,et al.  Centrifugal enhancement of retroviral mediated gene transfer. , 1995, Journal of virological methods.

[8]  I. Pastan,et al.  Retroviral coexpression of a multidrug resistance gene (MDR1) and human alpha-galactosidase A for gene therapy of Fabry disease. , 1995, Human gene therapy.

[9]  C. Tisher,et al.  Recurrent diseases in the kidney transplant. , 1994, American journal of kidney diseases : the official journal of the National Kidney Foundation.

[10]  J. Li,et al.  Transduction of CD34+ enriched cord blood and Gaucher bone marrow cells by a retroviral vector carrying the glucocerebrosidase gene. , 1994, Gene therapy.

[11]  S. Ishii,et al.  Human alpha-galactosidase gene expression: significance of two peptide regions encoded by exons 1-2 and 6. , 1994, Biochimica et biophysica acta.

[12]  R Kase,et al.  Characterization of a mutant alpha-galactosidase gene product for the late-onset cardiac form of Fabry disease. , 1993, Biochemical and biophysical research communications.

[13]  K. Cornetta,et al.  Infection of human cells with murine amphotropic replication-competent retroviruses. , 1993, Human gene therapy.

[14]  M. Sands,et al.  Reversal of pathology in murine mucopolysaccharidosis type VII by somatic cell gene transfer , 1992, Nature.

[15]  J. Li,et al.  Efficient transfer and sustained high expression of the human glucocerebrosidase gene in mice and their functional macrophages following transplantation of bone marrow transduced by a retroviral vector. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[16]  R. Mulligan,et al.  Safe and efficient generation of recombinant retroviruses with amphotropic and ecotropic host ranges. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Scheerer,et al.  Endocytosis of lysosomal alpha-galactosidase A by cultured fibroblasts from patients with Fabry disease. , 1982, American journal of human genetics.

[18]  J. Scheerer,et al.  Differential assay for lysosomal alpha-galactosidases in human tissues and its application to Fabry's disease. , 1981, Clinica chimica acta; international journal of clinical chemistry.

[19]  K. J. Dean,et al.  Enzyme therapy in Fabry disease: differential in vivo plasma clearance and metabolic effectiveness of plasma and splenic alpha-galactosidase A isozymes. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Brady,et al.  Replacement therapy for inherited enzyme deficiency. Use of purified ceramidetrihexosidase in Fabry's disease. , 1973, The New England journal of medicine.