Abstract

The long range objective of this study is the development of gene therapy for treatment of hereditary disorders due to missing or defective clotting factors. Gene therapy would involve genetic modification of some of a patient's somatic cells so that they secrete a continuing supply of clotting factor and thus effect long-term cure of the disease. This project will develop the methodology in animal models using factors VIII and IX as model genes, retroviral vectors for gene transfer, and skin fibroblasts, vascular smooth muscle cells, and skeletal muscle cells as gene transfer targets. The goal of the project is to demonstrate production of levels of functional human clotting factors in the animal's circulation that would be curative if achieved in human patients. Initial testing of the techniques will be performed in mice and rats, with subsequent studies in hemophiliac dogs. Of particular concern is the development of techniques that would be suitable for use in humans, and efforts to be sure that there be no deleterious effects that would limit their use in humans.
The specific aims i nclude the development of retroviral vectors for efficient gene expression, testing of various somatic cell types and methods for gene introduction, examination of the duration of clotting factor synthesis from the genetically-modified cells, and the study of possible adverse effects of the procedures. It is suggested that the techniques developed in this project might have applications in the 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 #
2R01HL041212-04
Application #
3358808
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1988-12-01
Project End
1995-11-30
Budget Start
1992-02-01
Budget End
1992-11-30
Support Year
4
Fiscal Year
1992
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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