The long-term objective of this proposal is to understand the molecular mechanisms which regulate the growth and differentiation of human epidermis. The health-relatedness of this objective lies in the use of the information acquired to design rational therapy for skin disease and to increase our understanding of skin disease pathogenesis. The main purpose of the proposed studies is to test crucial elements of two related hypotheses: A) that transforming growth factor-alpha (TGF-a) regulates the expression of keratinocyte protooncogenes, cytokines, and other mediators of proliferation and inflammation via signal transduction pathways known to be abnormal in psoriasis, and B) that TGF-alpha regulates a hyperproliferative mode of keratinocyte differentiation observed in cultured keratinocytes, wound healing, psoriasis and other epidermal hyperproliferative states.
The Specific Aims of this proposal are: 1) to define the role of the EGF receptor tyrosine kinase activity in the autoregulation of TGF-alpha, 2) to evaluate the effects of the KC antiproliferative agents transforming growth factor-Beta (TGF-Beta) and cyclosporin A (CsA) on TGF-alpha- induced KC phospholipid metabolism and gene expression, 3) to analyze the transcription and chromatin structure of the TGF-alpha gene, 4) To analyze the effects of transfected v-erb-B tyrosine kinase activity and antisense oligonucleotides to TGF-alpha on TGF-alpha gene expression and KC proliferation and 5) to analyze TGF-alpha gene expression in early lesions of psoriasis. These studies will be conducted in the Dermatology Research Unit of the University of Michigan on samples of normal and psoriatic human epidermis obtained by punch or keratome biopsy, on keratinocyte cultures obtained from these biopsies. The methods to be used include: RNA analysis by blot hybridization, nuclear runoff transcription, and in situ hybridization; lipid analysis by radiolabelling, organic solvent extraction, and immunofluorescence; chromatin structure analysis by nuclease digestion and blot hybridization, and gene transfer into keratinocytes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
1R29AR040016-01
Application #
3457297
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1989-07-01
Project End
1994-06-30
Budget Start
1989-07-01
Budget End
1990-06-30
Support Year
1
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Kansra, Sanjay; Stoll, Stefan W; Elder, James T (2002) Differential cytoskeletal association of ErbB1 and ErbB2 during keratinocyte differentiation. Biochem Biophys Res Commun 295:1108-17
Varani, J; Zeigler, M; Dame, M K et al. (2001) Heparin-binding epidermal-growth-factor-like growth factor activation of keratinocyte ErbB receptors Mediates epidermal hyperplasia, a prominent side-effect of retinoid therapy. J Invest Dermatol 117:1335-41
Stoll, S W; Kansra, S; Peshick, S et al. (2001) Differential utilization and localization of ErbB receptor tyrosine kinases in skin compared to normal and malignant keratinocytes. Neoplasia 3:339-50
Stoll, S W; Elder, J T (1999) Differential regulation of EGF-like growth factor genes in human keratinocytes. Biochem Biophys Res Commun 265:214-21
Stoll, S W; Elder, J T (1998) Retinoid regulation of heparin-binding EGF-like growth factor gene expression in human keratinocytes and skin. Exp Dermatol 7:391-7
Varani, J; Kang, S; Stoll, S et al. (1998) Human psoriatic skin in organ culture: comparison with normal skin exposed to exogenous growth factors and effects of an antibody to the EGF receptor. Pathobiology 66:253-9
Stoll, S W; Zhao, X; Elder, J T (1998) EGF stimulates transcription of CaN19 (S100A2) in HaCaT keratinocytes. J Invest Dermatol 111:1092-7
Stoll, S W; Benedict, M; Mitra, R et al. (1998) EGF receptor signaling inhibits keratinocyte apoptosis: evidence for mediation by Bcl-XL. Oncogene 16:1493-9
Xia, L; Stoll, S W; Liebert, M et al. (1997) CaN19 expression in benign and malignant hyperplasias of the skin and oral mucosa: evidence for a role in regenerative differentiation. Cancer Res 57:3055-62
Stoll, S; Garner, W; Elder, J (1997) Heparin-binding ligands mediate autocrine epidermal growth factor receptor activation In skin organ culture. J Clin Invest 100:1271-81

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