Perlecan is a large and complex protein composed of multiple modules which have growth-regulatory and cell adhesion among its properties. These studies are designed to characterize the heparan sulfate proteoglycan (HSPG) perlecan in the synovium at the molecular, biochemical and immunochemical levels and determine the gene expression and regulation of perlecan in normal synovial cells and synovial cells derived from rheumatoid arthritis (RA) and osteoarthritis (OA). The tissue-specific nature of synovial cell produced perlecan along with any differences in the gene expression and biochemical characteristics (i.e. size and side chain composition) of perlecan in synovium will be of fundamental importance to understanding the function of the synovium. A unique feature of this research proposal is that the synovium has been a tissue overlooked in regard to this proteoglycan primarily due to the normal and expected distribution of perlecan as part of the basement membrane. Without our recent cloning of the first human perlecan cDNA (and subsequent entire 14.5 kb cDNA), studies of perlecan mRNA using specific human perlecan probes were not possible. In preliminary studies, we have identified that synovial cells express perlecan and that there is significantly less perlecan mRNA expressed in RA synovial cells in comparison to non-arthritic and OA derived synovial cells. Although the complex character of the perlecan core protein has been partly elucidated where specific domains could be involved in important biological functions such as binding growth factors and prolonging their half-life, the precise role perlecan occupies in the synovium is presently unknown. Proinflammatory cytokines, IL-1 and TNF-alpha as well as TGF-beta and FGF will be examined in this proposal for their effects on perlecan gene transcription and biosynthesis. The existence of alternatively spliced perlecan transcripts in synovial cells will be investigated and the effects of these cytokines on their expression determined. Since perlecan is one of the major components of the classic basement membrane and had not been identified in the synovium until these studies, its further characterization in the synovium will provide new and important information. Recent studies show there are shared structural features between synovium and basement membranes, therefore, it would seem likely that many of the functions would also be in common. The presence and character of perlecan in the synovium would be of basis importance to re- defining the structure and function of this specialized tissue. The realized goals of this proposal will substantially add to our understanding of the synovium in normal joint homeostasis and in arthritis-related pathology.

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 #
5R29AR042417-03
Application #
2390525
Study Section
Special Emphasis Panel (ZRG4-GMA-2 (01))
Project Start
1995-04-01
Project End
2000-03-31
Budget Start
1997-04-01
Budget End
1998-03-31
Support Year
3
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Thomas Jefferson University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
061197161
City
Philadelphia
State
PA
Country
United States
Zip Code
19107
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Fang, Q; Sun, Y Y; Cai, W et al. (2000) Cartilage-reactive T cells in rheumatoid synovium. Int Immunol 12:659-69
Richardson, D W; Dodge, G R (2000) Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes. Am J Vet Res 61:624-30
Dodge, G R; Diaz, A; Sanz-Rodriguez, C et al. (1998) Effects of interferon-gamma and tumor necrosis factor alpha on the expression of the genes encoding aggrecan, biglycan, and decorin core proteins in cultured human chondrocytes. Arthritis Rheum 41:274-83
Dodge, G R; Hawkins, D; Boesler, E et al. (1998) Production of cartilage oligomeric matrix protein (COMP) by cultured human dermal and synovial fibroblasts. Osteoarthritis Cartilage 6:435-40
Richardson, D W; Dodge, G R (1998) Molecular characteristics of equine stromelysin and the tissue inhibitor of metalloproteinase 1. Am J Vet Res 59:1557-62
Jimenez, S A; Ala-Kokko, L; Prockop, D J et al. (1997) Characterization of human type II procollagen and collagen-specific antibodies and their application to the study of human type II collagen processing and ultrastructure. Matrix Biol 16:29-39
Richardson, D W; Dodge, G R (1997) Cloning of equine type II procollagen and the modulation of its expression in cultured equine articular chondrocytes. Matrix Biol 16:59-64