The aim of the research is to identify a human beta globin gene which can be regulated in a normal fashion after introduction into hematopoietic cells by retroviral transduction. Such research could provide the basis for a protocol of somatic gene therapy for patients with the severe inherited hemolytic anemias thalassemia major and sickle cell disease. Human globin genes in various configurations will be added to replication-defective retroviruses which also carry the gene for a dominant selectable marker such as resistance to the neomycin analogue G418 or the gene for a mutant dihydrofolate reductase which confers resistance to methotrexate. Human marrow cells as well as marrow from two animal model systems (mouse and dog) will be infected with globin containing retroviruses and the developmental activation, tissue or cell specificity, ultimate level, and persistence of globin expression will be studied. Assays will focus on the structure of the transduced globin gene, its transcripts, the amount and quality of the globin chains produced, and the effects on erythroid and nonerythroid cells. Assays for helper virus will be performed to help assess the risk of viral-induced disease. Experiments in the mouse will test whether pluripotential stem cells can accept a human globin gene and the stringency with which the human gene is activated in different hematopoietic lineages. Studies in the dog will extend these results in a larger animal which is a realistic model for human bone marrow transplantation. Studies with cultured human marrow may reveal if a therapeutic transduced globin gene can be expressed appropriately, and help refine infection protocols and efficient assays which could become the basis for a somatic therapy scheme centered on viral infection of bone marrow as an autologous transplant.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
7R01HL037073-05
Application #
3352626
Study Section
Special Emphasis Panel (SRC (29))
Project Start
1986-09-30
Project End
1991-12-31
Budget Start
1990-09-30
Budget End
1991-12-31
Support Year
5
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Icos Corporation
Department
Type
DUNS #
City
Bothell
State
WA
Country
United States
Zip Code
98021
Gelinas, R; Frazier, A; Harris, E (1992) A normal level of beta-globin expression in erythroid cells after retroviral cells transfer. Bone Marrow Transplant 9 Suppl 1:154-7
Walters, M; Kim, C; Gelinas, R (1991) Characterization of a DNA binding activity in DNAse I hypersensitive site 4 of the human globin locus control region. Nucleic Acids Res 19:5385-93
Osborne, W R; Hock, R A; Kaleko, M et al. (1990) Long-term expression of human adenosine deaminase in mice after transplantation of bone marrow infected with amphotropic retroviral vectors. Hum Gene Ther 1:31-41
Rixon, M W; Harris, E A; Gelinas, R E (1990) Expression of the human gamma-globin gene after retroviral transfer to transformed erythroid cells. Biochemistry 29:4393-400
Gelinas, R; Novak, U (1990) Retroviral vectors for the beta-globin gene that demonstrate improved titer and expression. Ann N Y Acad Sci 612:427-41
Novak, U; Harris, E A; Forrester, W et al. (1990) High-level beta-globin expression after retroviral transfer of locus activation region-containing human beta-globin gene derivatives into murine erythroleukemia cells. Proc Natl Acad Sci U S A 87:3386-90
Forrester, W C; Novak, U; Gelinas, R et al. (1989) Molecular analysis of the human beta-globin locus activation region. Proc Natl Acad Sci U S A 86:5439-43
Gelinas, R E; Bender, M A; Miller, A D et al. (1989) Long-term expression of the human beta-globin gene after retroviral transfer into pluripotent hematopoietic stem cells of the mouse. Adv Exp Med Biol 271:135-48
Gelinas, R E; Bender, M A; Miller, A D (1989) Regulated expression of the human beta-globin gene after retroviral transfer into murine and human hematopoietic cells. Prog Clin Biol Res 316B:235-49
Bender, M A; Gelinas, R E; Miller, A D (1989) A majority of mice show long-term expression of a human beta-globin gene after retrovirus transfer into hematopoietic stem cells. Mol Cell Biol 9:1426-34

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