The proposed program project will study various aspects of the molecular and cellular biology of various basement membrane zones of the skin, including the epidermal-dermal interface and the vascular basement membrane zone. The goals of this project focus on characterizing expression and regulation of basement membrane zone proteins of importance in pathophysiologic events involved in both normal physiology and epidermolysis bullosa (EB). We shall attempt to define structures and proteins involved in the functional integrity of the basement membrane zone and to characterize mechanisms governing their synthesis, secretion and degradation. The specific projects are as follows. I. """"""""Candidate Protein Clones in EB"""""""" will develop and use probes to map genes and establish polymorphisms in hereditary EB and define possible relationships between acquired EB and HLA. II. """"""""Keratinocyte Locomotion and Protease Expression"""""""" will examine the attachment and migratory behavior of keratinocytes and characterize the repertoire of metalloproteases expressed by the migrating cells. III. """"""""Metabolism of the Dermal Microvascular Basement Membrane"""""""" will examine matrix protein synthesis by normal and activated endothelial cells and compare the behavior of these cells with that of endothelial cells from patients with EB. IV. """"""""Regulation of Expression of Extracellular Matrix Molecules and Degradative Enzymes in Normal and RDEB Fibroblasts in Monolayer Culture and in a Reconstituted Skin Model"""""""" is predicated on the notion that excessive proteolysis is fundamental to understanding the connective tissue destruction that characterizes RDEB. It will explore the mechanisms of regulation of metalloprotease expression in a 3-dimensional model of reconstituted skin. V. """"""""Biology of the Interaction of Keratinocytes with Basement Membranes"""""""" will determine what role moesin, a heparin-binding receptor protein, may play in processes of cell-cell and cell-matrix interactions in cell spreading, growth and motility of the epidermis during development and wound healing in both normal and EB- derived cells. Ultimately, we hope that these studies will lead to better understanding of the function of the basement membrane zone and to more effective therapeutic approaches to EB.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Program Projects (P01)
Project #
5P01AR041045-04
Application #
2080432
Study Section
Arthritis and Musculoskeletal and Skin Diseases Special Grants Review Committee (AMS)
Project Start
1992-09-30
Project End
1997-08-31
Budget Start
1995-09-01
Budget End
1996-08-31
Support Year
4
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Stanford University
Department
Dermatology
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
Chen, Mei; O'Toole, Edel A; Sanghavi, Jigisha et al. (2002) The epidermolysis bullosa acquisita antigen (type VII collagen) is present in human colon and patients with crohn's disease have autoantibodies to type VII collagen. J Invest Dermatol 118:1059-64
Lara-Pezzi, E; Serrador, J M; Montoya, M C et al. (2001) The hepatitis B virus X protein (HBx) induces a migratory phenotype in a CD44-dependent manner: possible role of HBx in invasion and metastasis. Hepatology 33:1270-81
Yurko, M A; O'Toole, E A; Woodley, D T (2001) Phosphorylation of focal adhesion kinase (pp125(FAK)) is increased in human keratinocytes induced to migrate by extracellular matrices. J Cell Physiol 188:24-32
Wang, C K; Nelson, C F; Brinkman, A M et al. (2000) Spontaneous cell sorting of fibroblasts and keratinocytes creates an organotypic human skin equivalent. J Invest Dermatol 114:674-80
Nasca, M R; O'Toole, E A; Palicharla, P et al. (1999) Thalidomide increases human keratinocyte migration and proliferation. J Invest Dermatol 113:720-4
Chen, M; O'Toole, E A; Li, Y Y et al. (1999) Alpha 2 beta 1 integrin mediates dermal fibroblast attachment to type VII collagen via a 158-amino-acid segment of the NC1 domain. Exp Cell Res 249:231-9
Amieva, M R; Litman, P; Huang, L et al. (1999) Disruption of dynamic cell surface architecture of NIH3T3 fibroblasts by the N-terminal domains of moesin and ezrin: in vivo imaging with GFP fusion proteins. J Cell Sci 112 ( Pt 1):111-25
Chen, M; Marinkovich, M P; Jones, J C et al. (1999) NC1 domain of type VII collagen binds to the beta3 chain of laminin 5 via a unique subdomain within the fibronectin-like repeats. J Invest Dermatol 112:177-83
Nakamura, F; Huang, L; Pestonjamasp, K et al. (1999) Regulation of F-actin binding to platelet moesin in vitro by both phosphorylation of threonine 558 and polyphosphatidylinositides. Mol Biol Cell 10:2669-85
Matsui, C; Pereira, P; Wang, C K et al. (1998) Extent of laminin-5 assembly and secretion effect junctional epidermolysis bullosa phenotype. J Exp Med 187:1273-83

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