The goal of this proposal is to use genetically engineered myoblasts as vehicles for gene delivery for the treatment of immune deficiency disorders and cancer. Our approach capitalizes on a unique characteristic of muscle precursor cells. Myoblasts introduced into muscle by injection cross the basal lamina and fuse into pre-existing mature myofibers that are innervated and vascularized. As a result, injected myoblasts are not merely tolerated but nurtured, leading to longterm retention. We have previously demonstrated that myoblasts genetically engineered to express human growth hormone (hGH) stably deliver the hormone to the circulation for a period of three months. We now propose to use myoblast-mediated gene transfer to deliver two recombinant hematopoietic colony stimulating factors, M-CSF and GM-CSF. To date, therapeutic effects have not been reported using cell-mediated gene therapy. Initial studies will focus on gene transfer to correct a mouse model with a genetic defect. The osteopetrotic (op) mouse has a mutation within the M-CSF gene abrogating expression of the M-CSF gene and protein, leading to a severe deficiency in cells of the osteoclast and macrophage lineage. Implantation of genetically engineered myogenic cells into muscle of genetically defective animals will determine whether adequate levels of functional cytokine result from M-CSF gene transfer. Subsequent studies will determine whether myoblast-mediated delivery of CSFs is useful in overcoming dose-limiting toxicity resulting from chemotherapy in a mouse tumor model. Currently the major limitation to using higher doses of chemotherapy to kill cancer cells is the toxicity to bone marrow. The ability to achieve long-term systemic expression of modulators of immune and hematopoietic systems by gene therapy should have a profound impact on the clinical management of acquired diseases such as cancer and AIDS.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA059717-01
Application #
3203488
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1993-04-13
Project End
1998-03-31
Budget Start
1993-04-13
Budget End
1994-03-31
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Ozawa, Clare R; Banfi, Andrea; Glazer, Nicole L et al. (2004) Microenvironmental VEGF concentration, not total dose, determines a threshold between normal and aberrant angiogenesis. J Clin Invest 113:516-27
von Degenfeld, Georges; Banfi, Andrea; Springer, Matthew L et al. (2003) Myoblast-mediated gene transfer for therapeutic angiogenesis and arteriogenesis. Br J Pharmacol 140:620-6
Springer, Matthew L; Ozawa, Clare R; Banfi, Andrea et al. (2003) Localized arteriole formation directly adjacent to the site of VEGF-induced angiogenesis in muscle. Mol Ther 7:441-9
Brazelton, Timothy R; Nystrom, Michael; Blau, Helen M (2003) Significant differences among skeletal muscles in the incorporation of bone marrow-derived cells. Dev Biol 262:64-74
Wehrman, Tom; Kleaveland, Benjamin; Her, Jeng-Horng et al. (2002) Protein-protein interactions monitored in mammalian cells via complementation of beta -lactamase enzyme fragments. Proc Natl Acad Sci U S A 99:3469-74
Springer, Matthew L; Ozawa, Clare R; Blau, Helen M (2002) Transient production of alpha-smooth muscle actin by skeletal myoblasts during differentiation in culture and following intramuscular implantation. Cell Motil Cytoskeleton 51:177-86
Munz, Barbara; Hildt, Eberhard; Springer, Matthew L et al. (2002) RIP2, a checkpoint in myogenic differentiation. Mol Cell Biol 22:5879-86
Blau, H M; Brazelton, T R; Weimann, J M (2001) The evolving concept of a stem cell: entity or function? Cell 105:829-41
Blanco-Bose, W E; Blau, H M (2001) Laminin-induced change in conformation of preexisting alpha7beta1 integrin signals secondary myofiber formation. Dev Biol 233:148-60
Blanco-Bose, W E; Yao, C C; Kramer, R H et al. (2001) Purification of mouse primary myoblasts based on alpha 7 integrin expression. Exp Cell Res 265:212-20

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