The long-term goal of this study is to develop systems for foreign gene expression in rat airway epithelia in vivo, which can be applied to the development of gene therapy for cystic fibrosis. Specifically, the E.coli lacZ gene will be used as a reporter gene to design retroviral vectors that obtain the stable, long-term expression of beta-galactosidase in cultured airway epithelia from both rodents and humans. An in vivo epithelial culture system in denuded tracheas will be used to evaluate cell differentiation and longer-term gene expression. Systems will be develop to achieve foreign gene expression in intact airways of neonatal and adult rats. Cells transduced in vitro will be instilled into airways to determine if a grafting approach is feasible. In addition, direct vector infection of the airways will be attempted by instillation of vector stocks and by inhalation of aerosolized virus. Through these studies, we intend to determine the optimal parameters to achieve stable, high-level transgene expression in airway epithelia and to maximize the number of transduced cells in vivo.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL047119-03
Application #
3366321
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Project Start
1991-04-01
Project End
1995-03-31
Budget Start
1993-04-01
Budget End
1994-03-31
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Type
Schools of Medicine
DUNS #
077758407
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Yee, J K; Miyanohara, A; LaPorte, P et al. (1994) A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes. Proc Natl Acad Sci U S A 91:9564-8
Roemer, K; Friedmann, T (1992) Concepts and strategies for human gene therapy. Eur J Biochem 208:211-25