Long-term expression of the human beta-globin gene after retroviral transfer into pluripotent hematopoietic stem cells of the mouse.

Publication Type:

Journal Article

Source:

Advances in experimental medicine and biology, Volume 271, p.135-48 (1989)

Keywords:

1989, Animals, Bone Marrow Transplantation, Cells, Cultured, Chimera, Erythroid Precursor Cells, Gene Expression Regulation, GENES, Genetic Vectors, Globins, Graft Survival, Hematopoietic Stem Cells, Humans, Introns, Leukemia, Erythroblastic, Acute, MICE, Neoplastic Stem Cells, Recombinant Fusion Proteins, Retroviridae, Transfection, Tumor Cells, Cultured

Abstract:

We have studied the regulation of the human beta-globin gene after retroviral transfer into a variety of transformed and normal hematopoietic cells. After transfer into murine erythroleukemia cells (MEL) expression from the human beta-globin gene responds to inducers of erythroid maturation in parallel to the endogenous murine globin genes. After infection of human BFU-E, RNA expression from the virally-transferred beta-globin gene was measured at 2.5%-5% of the endogenous beta-globin level. The most improved globin vectors can transfer the human beta-globin gene into pluripotent hematopoietic stem cells in mouse bone marrow. Mice reconstituted with infected marrow show human beta-globin RNA and protein expression in peripheral blood cells for over 4 months. In these animals, both myeloid and lymphoid cells carry the integrated provirus at a level of about 1 copy per cell. In serial transplantation experiments, bone marrow from these animals is capable of repopulating secondary and tertiary recipient animals which go on to show long-term human beta-globin expression. Retroviral vectors thus provide a practical way to refine models of globin gene regulation through in vivo tests and to evaluate the feasibility of protocols for gene addition therapy.