The long term goal of this proposal is to develop methods for efficient delivery and expression of transgenes into keratinocytes for possible treatment of human diseases by somatic gene-based therapies. Ex vivo gene therapy involves removal of the target cells from body, transduction in vitro with viral vector and subsequent transplantation of the modified autologous cells back to the patient. This method requires surgical grafting, is time consuming and costly. Direct gene transfer to keratinocytes in epidermis would circumvent these difficulties. The objective of this research proposal is to develop a method to transduce basal keratinocytes in situ with retroviral vectors and achieve persistant presence of transgene in epidermal keratinocytes. This will be achieved by 1)induction of hyperplasia in epidermis to allow replication of basal keratinocytes required for retroviral integration, 2)determination of accessibility of basal cells to viral transduction using adenoviruses as a marker, 3)generation of very titer pseudotype retroviral vectors, 4)introduction of retroviral vectors to basal cells and determining persistency of the transgene in epidermis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32AR008390-02
Application #
2005987
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1996-10-01
Project End
Budget Start
1996-10-01
Budget End
1997-09-30
Support Year
2
Fiscal Year
1997
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
804878247
City
Stony Brook
State
NY
Country
United States
Zip Code
11794
Ghazizadeh, S; Taichman, L B (2001) Multiple classes of stem cells in cutaneous epithelium: a lineage analysis of adult mouse skin. EMBO J 20:1215-22
Ghazizadeh, S; Harrington, R; Taichman, L (1999) In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy. Gene Ther 6:1267-75
Ghazizadeh, S; Harrington, R; Garfield, J et al. (1998) Retrovirus-mediated transduction of porcine keratinocytes in organ culture. J Invest Dermatol 111:492-6