Our ultimate aim is the use of autotransplantation in combination with gene transfer for the cure of patients with homozygous Beta thalassemia and sickle cell anemia. The specific goal is to define the optimal conditions for the transfer and expression of human Beta globin genes into bone marrow cells. Studies to date have indicated that retroviral vectors can be used to efficiently transfer exogenous genes into mouse bone marrow cells. In addition, it has been shown that it is possible to express human Beta globin genes in erythroid cells in a tissue-specific manner. While other methods for treating patients with Beta thalassemia and sickle cell anemia exist, the potential use of gene transfer to treat these disorders is particularly exciting since it could provide a cure by appropriate expression of the normal Beta globin gene. In our initial studies, retroviral vectors containing the human Beta globin and the neomycin resistance genes will be used to transfer human Beta globin genes into mouse bone marrow cells, and expression assessed after transplantation into irradiated mice. These experiments will also test whether enhancer sequences derived from cloned DNA segments from within the Beta globin gene complex as well as from other sources can increase Beta globin expression. Amplification of integrated human Beta globin genes will be another method for attempting to increase Beta globin production. A vector containing a dihydrofolate reductase (DHFR) gene will be used for this purpose. A mutant mouse strain with Beta thalassemia as well as normal mice will be used to assess the expression of human Beta globin genes in intact mice. The anemia in these Beta thalassemic mice has already been corrected by a human Beta globin gene inserted transgenically in mouse embryos. In further experiments, when safe and appropriate amphotropic retroviral vectors become available, we plan to introduce the human Beta globin gene into monkey bone marrow cells. These experiments in primates will hopefully be the final step toward demonstrating the feasibility of effective gene therapy of human patients with Beta thalassemia and sickle cell anemia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL037069-01
Application #
3352613
Study Section
(SRC)
Project Start
1986-09-30
Project End
1991-09-29
Budget Start
1986-09-30
Budget End
1987-09-29
Support Year
1
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027
O'Neill, D; Kaysen, J; Donovan-Peluso, M et al. (1990) Protein-DNA interactions upstream from the human A gamma globin gene. Nucleic Acids Res 18:1977-82
Beldjord, C; Lapoumeroulie, C; Pagnier, J et al. (1988) A novel beta thalassemia gene with a single base mutation in the conserved polypyrimidine sequence at the 3' end of IVS 2. Nucleic Acids Res 16:4927-35
Markowitz, D; Goff, S; Bank, A (1988) A safe packaging line for gene transfer: separating viral genes on two different plasmids. J Virol 62:1120-4
Markowitz, D; Goff, S; Bank, A (1988) Construction of a safe and efficient retrovirus packaging cell line. Adv Exp Med Biol 241:35-40
Markowitz, D; Goff, S; Bank, A (1988) Construction and use of a safe and efficient amphotropic packaging cell line. Virology 167:400-6
Donovan-Peluso, M; Acuto, S; Swanson, M et al. (1987) Expression of human gamma-globin genes in human erythroleukemia (K562) cells. J Biol Chem 262:17051-7
Lerner, N; Brigham, S; Goff, S et al. (1987) Human beta-globin gene expression after gene transfer using retroviral vectors. DNA 6:573-82
Acuto, S; Donovan-Peluso, M; Giambona, N et al. (1987) The role of human globin gene promoters in the expression of hybrid genes in erythroid and non-erythroid cells. Biochem Biophys Res Commun 143:1099-106
Rund, D; Dobkin, C; Bank, A (1987) Regulated expression of amplified human beta globin genes. Blood 70:733-9
Bank, A; Donovan-Peluso, M; Lerner, N et al. (1987) Human globin gene expression after gene transfer. Blood Cells 13:269-75

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