Abstract

The long range objective of this study is to utilize """"""""gene therapy"""""""" for treatment of bleeding disorders that are the result of missing or defective clotting factors. Gene therapy would involve genetic modification of some of a patient's somatic cells to secrete a constant supply of clotting factor to effect long-term cure of the disease. This project will develop the methodology in an animal model by using factor IX as a model gene, retroviral vectors for gene transfer, and skin fibroblasts as gene transfer recipients. The goal of the project is to demonstrate production of circulating levels of active human factor IX in animals that would be curative if achieved in human patients. Of particular concern is that the techniques developed here would be suitable for use in humans, and that there be no deleterious effects that would limit their use in humans.
The specific aims i nvolve the development of retroviral vectors for efficient gene expression, testing of various methods for reintroduction of the genetically modified fibroblasts, examination of the persistence of and continued factor IX synthesis from the reintroduced fibroblasts, and study of possible adverse effects of the procedures. It is hoped the techniques developed here will have applications to treatment of genetic diseases in general.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL041212-01
Application #
3358807
Study Section
(SRC)
Project Start
1988-12-01
Project End
1991-11-30
Budget Start
1988-12-01
Budget End
1989-11-30
Support Year
1
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109

  Publications

Koeberl, D D; Alexander, I E; Halbert, C L et al. (1997) Persistent expression of human clotting factor IX from mouse liver after intravenous injection of adeno-associated virus vectors. Proc Natl Acad Sci U S A 94:1426-31
Adam, M A; Osborne, W R; Miller, A D (1995) R-region cDNA inserts in retroviral vectors are compatible with virus replication and high-level protein synthesis from the insert. Hum Gene Ther 6:1169-76
Koeberl, D D; Halbert, C L; Krumm, A et al. (1995) Sequences within the coding regions of clotting factor VIII and CFTR block transcriptional elongation. Hum Gene Ther 6:469-79
Russell, D W; Miller, A D; Alexander, I E (1994) Adeno-associated virus vectors preferentially transduce cells in S phase. Proc Natl Acad Sci U S A 91:8915-9
Alexander, I E; Russell, D W; Miller, A D (1994) DNA-damaging agents greatly increase the transduction of nondividing cells by adeno-associated virus vectors. J Virol 68:8282-7
Lynch, C M; Israel, D I; Kaufman, R J et al. (1993) Sequences in the coding region of clotting factor VIII act as dominant inhibitors of RNA accumulation and protein production. Hum Gene Ther 4:259-72
Palmer, T D; Miller, A D; Reeder, R H et al. (1993) Efficient expression of a protein coding gene under the control of an RNA polymerase I promoter. Nucleic Acids Res 21:3451-7
Palmer, T D; Rosman, G J; Osborne, W R et al. (1991) Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc Natl Acad Sci U S A 88:1330-4
Kaleko, M; Garcia, J V; Miller, A D (1991) Persistent gene expression after retroviral gene transfer into liver cells in vivo. Hum Gene Ther 2:27-32
Adam, M A; Ramesh, N; Miller, A D et al. (1991) Internal initiation of translation in retroviral vectors carrying picornavirus 5' nontranslated regions. J Virol 65:4985-90

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