Antisense inhibition of chondrocyte CD44 expression leading to cartilage chondrolysis

Chow, Geraldine; Nietfeld, J. J.; Knudson, Cheryl B.; Knudson, Warren

doi:10.1002/1529-0131(199808)41:8<1411::aid-art10>3.0.co;2-z

Cited by 63 publications

(49 citation statements)

References 37 publications

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“…In adult cartilage HA oligo activate a "matrix repair" response that includes both a massive loss of extracellular matrix components because of the activation of gelatinolytic enzymes and aggrecanase as well as the induction of new matrix biosynthesis. Similar results are obtained when cartilage is treated with antisense oligonucleotides directed against CD44 (33) or chondrocytes are treated with hyaluronidase (26). Thus interfering with CD44-HA interactions by perturbing either the HA or the CD44, both induce metabolic changes away from matrix homeostasis.…”

supporting

confidence: 77%

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Ohno

Knudson

et al. 2006

Journal of Biological Chemistry

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114

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Hyaluronan exerts a variety of biological effects on cells including changes in cell migration, proliferation, and matrix metabolism. However, the signaling pathways associated with the action of hyaluronan on cells have not been clearly defined. In some cells, signaling is induced by the loss of cell-hyaluronan interactions. The goal of this study was to use hyaluronan oligosaccharides as a molecular tool to explore the effects of changes in cell-hyaluronan interactions and determine the underlying molecular events that become activated. In this study, hyaluronan oligosaccharides induced the loss of extracellular matrix proteoglycan and collagen from cultured slices of normal adult human articular cartilage. This loss was coincident with an increased expression of matrix metalloproteinase (MMP)-13. MMP-13 expression was also induced in articular chondrocytes by hyaluronan (HA) hexasaccharides but not by HA tetrasaccharides nor high molecular weight hyaluronan. MMP-13 promoterreporter constructs in CD44-null COS-7 cells revealed that both CD44-dependent and CD44-independent events mediate the induction of MMP-13 by hyaluronan oligosaccharides. Electromobility gel shift assays demonstrated the activation of chondrocyte NFB by hyaluronan oligosaccharides. NFB activation was also documented in C-28/I2 immortalized human chondrocytes by luciferase promoter assays and phosphorylation of IKK-␣/␤. The link between activation of NFB and MMP-13 induction by HA oligosaccharides was further confirmed through the use of the NFB inhibitor helenalin. Inhibition of MAP kinases also demonstrated the involvement of p38 MAP kinase in the hyaluronan oligosaccharide induction of MMP-13. Our findings suggest that hyaluronan-CD44 interactions affect matrix metabolism via activation of NFB and p38 MAP kinase.In many tissues cell-matrix interactions serve to regulate cellular homeostasis (1, 2). Interference with these associations typically signal cells to initiate matrix repair, cell proliferation, or even cell migration. Such interactions are especially important in tissues such as articular cartilage, a tissue rich in extracellular matrix but with limited vascular access for systemic control of homeostasis. Chondrocytes are thus highly dependent on cell-matrix interactions as a primary means to "sense" changes in the extracellular environment (3). Most investigations in this area focus on cell signaling induced through the interaction of integrin receptors with collagens, fibronectin, laminin, matrilins, etc.(3-7). However, cell-matrix interactions involving hyaluronan (HA), 2 once thought to be predominately structural in nature, are now beginning to receive increasing attention as initiators of cell signaling.HA is a high molecular weight polysaccharide consisting of repeating disaccharide units glucuronic acid and N-acetylglucosamine (8). In cartilage and many other connective tissues, HA serves as a central filamentous scaffold to which proteoglycans such as aggrecan and link protein become bound (9). This complex matrix networ...

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supporting

confidence: 77%

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Ohno

Knudson

et al. 2006

Journal of Biological Chemistry

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114

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“…These results suggest that whatever mechanism was responsible for the activation, the pathway differed significantly from pathways activated by IL-1. A similar catabolic response was also obtained in bovine articular cartilage slices when CD44 expression was reduced through the use of small phosphorothioate antisense oligonucleotides (11). Thus, it would appear that interference with cell-matrix interactions, by both perturbation of ligand binding and receptor-protein biosynthesis lead to similar changes in chondrocyte metabolism.…”

supporting

confidence: 55%

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Ohno¹,

Im²,

Knudson³

et al. 2005

Arthritis & Rheumatism

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Objective. To document the activity profile of transcription factors following chondrocyte stimulation with hyaluronan (HA) hexasaccharides (HA 6 ) and to determine the expression of genes whose transcriptional activation is tightly associated with the transcription factors.Methods. Nuclear extracts from bovine articular chondrocytes treated with HA 6 were subjected to transcription factor protein-DNA array analysis. Electrophoretic mobility shift assay (EMSA) analyses were performed to confirm the results of protein-DNA array. The gene expressions of matrix metalloproteinase 3 (MMP-3), type II collagen, and cartilage oligomeric matrix protein (COMP) were examined by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR), and protease activity was assessed by casein zymography.Results. In the protein-DNA array analysis, 12 transcription factors were up-regulated and 2 transcription factors were down-regulated in the chondrocytes treated with HA 6 . The transcription factors retinoic acid receptor (RAR), retinoid X receptor (RXR), and Sp-1 exhibited >2-fold increased activity by HA 6 treatment, as confirmed by EMSA. RT-PCR analysis showed that the expression levels of MMP-3, type II collagen, and COMP messenger RNA, which are tightly associated with the activation of RAR, RXR, or Sp-1, were upregulated by treatment with HA 6 . Addition of high molecular mass HA after HA 6 treatment resulted in abrogation of the MMP-3 induction. Conclusion.These results suggest that HA 6 increase the activity of multiple transcription factors in chondrocytes and signal the enhanced expression of key genes involved in cartilage-matrix remodeling and turnover. The data also demonstrate that high molecular mass HA has a potential to suppress the signaling activated by HA 6 .The glycosaminoglycan hyaluronan (HA) is a critical component of the extracellular matrix of articular cartilage. HA serves as a scaffold for the aggregation of cartilage proteoglycan (1) and, in this manner, forms a continuum of structure throughout the extracellular matrix. In addition, we have shown that HA facilitates the anchorage of these proteoglycan aggregates to the chondrocyte cell surface via its interaction with the membrane HA receptor CD44 (2-4). Thus, it is, in part, the interaction of chondrocytes directly with HA that constitutes the cell-matrix interaction that serves to regulate many aspects of chondrocyte metabolism (5). We have shown previously that one method of interference with the cell-matrix interactions of chondrocytes is through the use of excess small HA oligosaccharides, such as HA hexasaccharides (HA 6 ) (2,4,6). HA 6 compete with high molecular mass HA for CD44 binding; yet, they are too small in size to interfere with HA aggrecan or HA-link protein interactions (1,7,8).The consequences of this loss of cell-matrix interactions can be dramatic. In a previous study, we demonstrated that the addition of small HA oligosaccharides to human or bovine articular cartilage slices resulted in the apparent activation o...

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“…The interaction between HA and CD44 has been shown to reduce anti-Fas-induced apoptosis of chondrocytes (36), which is further evidence of the direct involvement of CD44 in the mechanism of HA action. Antisense inhibition of chondrocyte CD44 expression results in proteoglycan degradation with NITEGE expression in cartilage (37), indicating that CD44 may be a key player in the maintenance of the cartilage structure. Of interest, monovalent ligation of the anti-CD44 antibody IM7 with CD44 also caused significant inhibitory effects on IL-1␤-stimulated MMPs (Figures 5, 6, and 7), which reflects a minor role of the barrier effect of HA through pericellular accumulation around chondrocytes.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of interleukin‐1β‐stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage

Julovi

Yasuda

Shimizu

et al. 2004

Arthritis & Rheumatism

156

122

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Objective. To investigate the mechanism of the inhibitory action of hyaluronan (HA) on interleukin-1␤ (IL-1␤)-stimulated production of matrix metalloproteinases (MMPs) in human articular cartilage.Methods. IL-1␤ was added to normal and osteoarthritic (OA) human articular cartilage in explant culture to stimulate MMP production. Articular cartilage was incubated or preincubated with a clinically used form of 800-kd HA to assess its effect on IL-1␤-induced MMPs. Levels of secreted MMPs 1, 3, and 13 in conditioned media were detected by immunoblotting; intracellular MMP synthesis in chondrocytes was evaluated by immunofluorescence microscopy. Penetration of HA into cartilage tissue and its binding to CD44 were analyzed by fluorescence microscopy using fluoresceinated HA. Blocking experiments with anti-CD44 antibody were performed to investigate the mechanism of action of HA.Results. Treatment and pretreatment with 800-kd HA at 1 mg/ml resulted in significant suppression of IL-1␤-stimulated production of MMPs 1, 3, and 13 in normal and OA cartilage explant culture. Fluorescence histocytochemistry revealed that HA penetrated cartilage tissue and localized in the pericellular matrix around chondrocytes. HA-binding blocking experiments using anti-CD44 antibody demonstrated that the association of HA with chondrocytes was mediated by CD44. Preincubation with anti-CD44 antibody, which suppressed IL-1␤-stimulated MMPs, reversed the inhibitory effect of HA on MMP production that was induced by IL-1␤ in normal and OA cartilage.Conclusion. This study demonstrates that HA effectively inhibits IL-1␤-stimulated production of MMP-1, MMP-3, and MMP-13, which supports the clinical use of HA in the treatment of OA. The action of HA on IL-1␤ may involve direct interaction between HA and CD44 on chondrocytes.Osteoarthritis (OA) is the most prevalent disease of articular joints and is the major cause of disability in the elderly. Pathophysiologic changes occur in OA cartilage due to the excessive expression of cartilagedegrading proteinases, the resultant progressive breakdown of collagen fibers, and the degradation of proteoglycan, mainly aggrecan (1).Matrix metalloproteinases (MMPs) are zinccontaining, calcium-dependent proteinases, which collectively degrade all components of the extracellular matrix. MMPs are considered to be important in the chondrolytic processes that contribute to the degenerative changes in OA cartilage (2-4). Recent studies have identified the messenger RNA (mRNA) for some MMPs, such as MMP-1, in human OA cartilage (4,5), and other investigators have reported specific MMP proteins and collagenasemediated type II collagen degradation products (6,7). There is a consensus that these enzymes play a critical role in intrinsic chondrocyte-mediated degenerative changes of the cartilage matrix in OA. Proinflammatory cytokines such as interleukin-1 (IL-1) strongly stimulate the expression of MMPs by chondrocytes in arthritis (8).Hyaluronan (HA) is a major component of synovial fluid and cartilage matrix, and it play...

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Antisense inhibition of chondrocyte CD44 expression leading to cartilage chondrolysis

Abstract: CD44 expression is needed for maintenance of cartilage homeostasis.

Cited by 63 publications

References 37 publications

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Hyaluronan Oligosaccharides Induce Matrix Metalloproteinase 13 via Transcriptional Activation of NFκB and p38 MAP Kinase in Articular Chondrocytes

Hyaluronan oligosaccharide–induced activation of transcription factors in bovine articular chondrocytes

Inhibition of interleukin‐1β‐stimulated production of matrix metalloproteinases by hyaluronan via CD44 in human articular cartilage

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