Collagenase is critical in initiating the degradation of interstitial collagens in a variety of morphogenetically important events. We shall use cultured skin fibroblasts to examine the effects of collagenase-stimulatory proteins, including a 20 kDa epidermal cytokine and platelet-derived growth factor, on the expression of collagenase by these target cells. We shall assess whether the response to these peptide agonists is developmentally modulated by examining (i) collagenase expression as reflected by enzyme activity, synthesis of immunoreactive enzyme protein and alterations in hybridizable mRNA and (ii) the mitogenic response of the target fibroblasts to the cytokines. We shall use cultured fibroblasts from various stages of development including from the genetic """"""""aging syndromes"""""""" to address whether these two responses are reflective of separate mechanistic pathways. These investigations of the mechanisms for the effects of cytokines on collagenase expression will be complemented by defining if the gene product itself, collagenase, can in any way affect its own synthesis, by probing the effects of other agonists on normal cells and cells from various types of epidermolysis bullosa, and by examining the effects at a nucleic acid level of agents such as retinoids and glucocorticoids that inhibit collagenase synthesis. Since there is evidence for a structurally mutant form of collagenase in recessive dystrophic epidermolysis bullosa (RDEB), we shall employ cultured fibroblasts from this disease as a source for collagenase mRNA to clone the putatively mutant collagenase gene. As a probe for the isolation and initial characterization of the RDEB cDNa clones we shall employ a full-length cDNA clone for normal human skin collagenase pCol 185.2. Our goal is to focus intensely on defining the site(s) of mutation leading to the structurally altered protein, so that we can better understand regulatory mechanisms, provide improved diagnosis and genetic counseling, and devise rational therapeutic modalities in this devastating genodermatosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37AR019537-15
Application #
3481480
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1988-06-01
Project End
1991-11-30
Budget Start
1989-12-01
Budget End
1990-11-30
Support Year
15
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Stanford University
Department
Type
Schools of Medicine
DUNS #
800771545
City
Stanford
State
CA
Country
United States
Zip Code
94305
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
Oh, D H; Stanley, R J; Lin, M et al. (1997) Two-photon excitation of 4'-hydroxymethyl-4,5',8-trimethylpsoralen. Photochem Photobiol 65:91-5
Lin, M; Hultquist, K L; Oh, D H et al. (1995) Inhibition of collagenase type I expression by psoralen antisense oligonucleotides in dermal fibroblasts. FASEB J 9:1371-7
Matsui, C; Wang, C K; Nelson, C F et al. (1995) The assembly of laminin-5 subunits. J Biol Chem 270:23496-503
Hoeffler, W K; Levinson, A D; Bauer, E A (1994) Activation of c-Jun transcription factor by substitution of a charged residue in its N-terminal domain. Nucleic Acids Res 22:1305-12
McGowan, K A; Bauer, E A; Smith, L T (1994) Localization of type I human skin collagenase in developing embryonic and fetal skin. J Invest Dermatol 102:951-7
Unemori, E N; Mauch, C; Hoeffler, W et al. (1994) Constitutive activation of the collagenase promoter in recessive dystrophic epidermolysis bullosa fibroblasts: role of endogenously activated AP-1. Exp Cell Res 211:212-8
Ceilley, E; Watanabe, N; Shapiro, D et al. (1993) Labeling of fractured human skin with antibodies to BM 600/nicein, epiligrin, kalinin and other matrix components. J Dermatol Sci 5:97-103
Unemori, E N; Beck, L S; Lee, W P et al. (1993) Human relaxin decreases collagen accumulation in vivo in two rodent models of fibrosis. J Invest Dermatol 101:280-5
Krejci, N C; Knapp, D M; Rudd, R J et al. (1992) Dermal mast cell granules bind interstitial procollagenase and collagenase. J Invest Dermatol 98:748-52

Showing the most recent 10 out of 25 publications