We propose to utilize the knowledge and expertise we have acquired in our ongoing studies of viral-keratinocyte interactions to explore the use of keratinocytes as vehicles for gene therapy. Many of the potential complications associated with the use of marrow cells in gene therapy would be reduced or eliminated with the use of genetically altered keratinocytes. What is needed for a proper consideration of this concept is an understanding of exogenous gene expression in keratinocytes. Preliminary studies have shown that high level transient expression of the gene for chloramphenicol acetyltransferase is possible in cultured epidermal keratinocytes following DNA-mediated gene transfer. In this proposal, we have set forth experiments with cloned genes and cultured human epidermal keratinocytes that are designed to (1) explore the use of calcium phosphate cotransfection and an adenovirus vector system as methods to achieve stable gene transfer and expression, (2) examine the effect of keratinocyte differentiation on exogenous gene expression, (3) investigate the effect of exogenous gene expression upon the growth and differentiation of the genetically-transformed keratinocyte, (4) explore the possibility of using a segment of adenovirus in the construction of a differentiation-activated expression vector, and (5) determine, with an experimental animal model, if genetically altered keratinocytes are a feasible means of administering gene therapy. In addition to providing vital information on the possibility of using keratinocytes for gene therapy, these studies will continue to help us gain an understanding of how keratinization in oral mucosa and epidermis is regulated.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE004511-12
Application #
3219086
Study Section
Oral Biology and Medicine Study Section (OBM)
Project Start
1979-05-01
Project End
1989-04-30
Budget Start
1987-05-01
Budget End
1988-04-30
Support Year
12
Fiscal Year
1987
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Type
Schools of Dentistry/Oral Hygn
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Ghazizadeh, Soosan; Taichman, Lorne B (2005) Organization of stem cells and their progeny in human epidermis. J Invest Dermatol 124:367-72
Ghazizadeh, Soosan; Katz, Anne B; Harrington, Robin et al. (2004) Lentivirus-mediated gene transfer to human epidermis. J Investig Dermatol Symp Proc 9:269-75
Ghazizadeh, Soosan; Kalish, Richard S; Taichman, Lorne B (2003) Immune-mediated loss of transgene expression in skin: implications for cutaneous gene therapy. Mol Ther 7:296-303
Ghazizadeh, S; Doumeng, C; Taichman, L B (2002) Durable and stratum-specific gene expression in epidermis. Gene Ther 9:1278-85
Albers, K M; Greif, F; Setzer, R W et al. (1987) Cell-cycle withdrawal in cultured keratinocytes. Differentiation 34:236-40