The long term objective of this research is to achieve successful gene therapy of human diseases of the hematopoietic system, specifically of the beta thalassemias and sickle cell anemia.
Our specific aims are: to design new retroviral vectors for efficient selection of infected totipotent hematopoietic stem cells so as to achieve 100 percent long-term reconstitution of transplanted animals and complete elimination of non- infected cells; to infect human and murine totipotent hematopoietic stem cells from normal and beta-thalassemic subjects with optimized retroviral vectors for transferring human beta globin gene and Locus Control Region derivatives and to test their in vivo properties; and to transfer a globin gene by retroviral vectors for gene therapy of sickle cell disease. These pre-clinical studies are designed to provide the scientific and technologic basis for subsequent clinical studies of gene therapy of these hematologic disorders. This proposal represents a collaboration between Irving M. London, Philippe LeBoulch and Dorothy Tuan of the Harvard-MIT Division of Health Sciences and Technology of MIT, Connie Eaves and R. Keith Humphries of the Terry Fox Laboratories of the British Columbia Cancer Agency, and Yves Beuzard of the University of Paris XII.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL048374-04
Application #
2224441
Study Section
Special Emphasis Panel (SRC (FJ))
Project Start
1992-05-01
Project End
1997-02-28
Budget Start
1995-03-01
Budget End
1996-02-29
Support Year
4
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Massachusetts Institute of Technology
Department
Type
Organized Research Units
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139
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Migliaccio, A R; Bengra, C; Ling, J et al. (2000) Stable and unstable transgene integration sites in the human genome: extinction of the Green Fluorescent Protein transgene in K562 cells. Gene 256:197-214
Pawliuk, R; Bachelot, T; Raftopoulos, H et al. (1998) Retroviral vectors aimed at the gene therapy of human beta-globin gene disorders. Ann N Y Acad Sci 850:151-62
Pawliuk, R; Eaves, C J; Humphries, R K (1997) Sustained high-level reconstitution of the hematopoietic system by preselected hematopoietic cells expressing a transduced cell-surface antigen. Hum Gene Ther 8:1595-604
Bouhassira, E E; Westerman, K; Leboulch, P (1997) Transcriptional behavior of LCR enhancer elements integrated at the same chromosomal locus by recombinase-mediated cassette exchange. Blood 90:3332-44
Kong, S; Bohl, D; Li, C et al. (1997) Transcription of the HS2 enhancer toward a cis-linked gene is independent of the orientation, position, and distance of the enhancer relative to the gene. Mol Cell Biol 17:3955-65
Westerman, K A; Leboulch, P (1996) Reversible immortalization of mammalian cells mediated by retroviral transfer and site-specific recombination. Proc Natl Acad Sci U S A 93:8971-6
Takekoshi, K J; Oh, Y H; Westerman, K W et al. (1995) Retroviral transfer of a human beta-globin/delta-globin hybrid gene linked to beta locus control region hypersensitive site 2 aimed at the gene therapy of sickle cell disease. Proc Natl Acad Sci U S A 92:3014-8
Leboulch, P; Huang, G M; Humphries, R K et al. (1994) Mutagenesis of retroviral vectors transducing human beta-globin gene and beta-globin locus control region derivatives results in stable transmission of an active transcriptional structure. EMBO J 13:3065-76