Long-term expression of retroviral-transduced adenosine deaminase in human primitive hematopoietic progenitors.
Adenosine deaminase (ADA) deficiency, a rare autosomal recessive disorder, is an ideal candidate for gene replacement therapy. By means of co-cultivation with a retroviral vector-producing cell line, we have demonstrated efficient transfer and expression of the human ADA gene into human primitive hematopoietic progenitors. At 6 weeks post-transduction in myeloid long-term bone marrow culture, approximately 50% of the clonogenic progenitors were transduced by the provirus, with ADA expression detected in 30% of transduced colonies. The ADA activity increased by 3.7-fold in the nonadherent fraction of transduced bone marrow after 9 weeks. We have also achieved efficient transduction by retroviral supernatant of normal and ADA-deficient bone marrow cells that were allowed to establish a stromal layer in long-term culture, indicating the feasibility of proceeding with attempts to perform stem cell gene therapy on patients with ADA deficiency.
Mitani
K
K
Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030.
Wakamiya
M
M
Caskey
C T
CT
D016475:3T3 Cells; D000243:Adenosine Deaminase / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells; D004277:DNA, Single-Stranded; D015870:Gene Expression; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000201:enzymology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics*; D017154:Stromal Cells / Q000378:metabolism; D014161:Transduction, Genetic
D004277:DNA, Single-Stranded; D012333:RNA, Messenger; D012367:RNA, Viral; D000243:Adenosine Deaminase