Aggrecanolysis in human osteoarthritis: confocal localization and biochemical characterization of ADAMTS5–hyaluronan complexes in articular cartilages

Plaas, Anna; Osborn, Barbara; Yoshihara, Yasuo; Bai, Yun; Bloom, Timothy J.; Nelson, Fred R.T.; Mikecz, Katalin; Sandy, John D.

doi:10.1016/j.joca.2006.12.008

Cited by 101 publications

(99 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Fivemicron sections were cut and stained with Masson's trichrome staining (Electron Microscopy Sciences, Hatfield, PA) according to the manufacturer's instructions. Confocal immunohistochemistry was performed essentially as described previously (33). Briefly, sections were deparaffinized and incubated with 1 g/ml of affinity-purified antibody, ␣-DLS, to detect aggrecan (33), rabbit IgG for nonimmune control (supplemental Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Confocal immunohistochemistry was performed essentially as described previously (33). Briefly, sections were deparaffinized and incubated with 1 g/ml of affinity-purified antibody, ␣-DLS, to detect aggrecan (33), rabbit IgG for nonimmune control (supplemental Fig. S2), ab1033 for versican ␤-GAG domain (Abcam), or biotinylated aggrecan-G1 domain for hyaluronan (supplemental Fig.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, by degrading pericellularly localized aggrecan, it appears to maintain pro-fibrotic TGF␤1 signaling in fibroblastic cells. It is likely that this unique function of TS5 is related to its confinement to the HA-rich pericellular matrix of cells in nonhealing ligaments (31) and in the superficial layer of articular cartilage (33).…”

Section: Finally In Both Cd44mentioning

confidence: 99%

See 2 more Smart Citations

Adamts5 Deletion Blocks Murine Dermal Repair through CD44-mediated Aggrecan Accumulation and Modulation of Transforming Growth Factor β1 (TGFβ1) Signaling

Velasco

DiPietro

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

ADAMTS5 has been implicated in the degradation of cartilage aggrecan in human osteoarthritis. Here, we describe a novel role for the enzyme in the regulation of TGF␤1 signaling in dermal fibroblasts both in vivo and in vitro. Dermal wound healing involves a series of cellular responses that depend on the coordinated temporal and spatial expression of inflammatory cytokines, proteases, growth factors, and extracellular matrix molecules. The early phases of hemostasis, inflammation, and re-epithelialization are followed by granulation tissue formation and finally dermal remodeling (1, 2). The granulation tissue is a loosely organized collagenous matrix that serves as a template for cellular regeneration (3) of the collagen (4) and proteoglycan (5-7) structures of the dermis proper.There has been a long held consensus that proteinases, such as plasmin and matrix metalloproteinases, are major players in dermal repair; however, many questions remain regarding substrate and product identification and to what extent cleavage of particular proteins is destructive or reparative (reviewed in Ref. 8). Although much interest has focused on matrix metalloproteinases (9), the ADAMTSs have now also been found to play a major role in a range of repair and regeneration processes. For example, evidence of a role for ADAMTS activity in dermal repair comes from the finding that human skin contains abundant aggrecanase-generated catabolic fragments of versican V0 and V1 (5) and that both aggrecan and versican are associated with the dermal region of the hair follicle (7). Moreover, to place dermal matrix turnover in a broader context, cleavage of the hyalectans aggrecan, versican, and brevican by one or more aggrecanases (ADAMTS1, -4, -5, -8, -9, and -15) occurs in the remodeling of cartilage (10), aorta (11, 12), spinal cord (13), meniscus (14, 15) intervertebral disc (16), adipose tissue (17), and brain (18). HA 2 in the skin is most abundant in the epidermal layer (6), and it undergoes partial fragmentation in both the dermis and epidermis of human skin explants (19). Its role in skin biology has largely focused on its binding to the cell-surface receptor CD44, an interaction that prevents PDGF-BB receptor activation and human fibroblast migration (20). It also appears to be required for MMP9-mediated activation of TGF␤1 (21). HA has been shown to ligate RHAMM to promote the association of CD44 and p-ERK, an interaction that directly activates cell migration and thereby is required for efficient dermal repair (22,23). HA also regulates the proliferative response of dermal fibroblasts to TGF␤1 (24), and HA12 oligosaccharides stimulate Col3 and TGF␤1 expression (25), a finding that is very relevant to the data described here.TGF␤1 is a key regulator of multiple processes in dermal wound healing (26). In the context of this study, TGF␤1 stimulates col1 and col3 production by fibroblasts and is required for terminal differentiation of myofibroblasts during wound * This work was supported, in whole or in part, by National Institutes o...

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Finally In Both Cd44mentioning

confidence: 99%

See 1 more Smart Citation

Adamts5 Deletion Blocks Murine Dermal Repair through CD44-mediated Aggrecan Accumulation and Modulation of Transforming Growth Factor β1 (TGFβ1) Signaling

Velasco

DiPietro

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…are known to mediate binding to cell surface glycosaminoglycans (19,20), as well as play an important role in substrate recognition and binding (18,19,23). Hyaluronan binding to the Dis domain (20) and binding of heparan sulfate proteoglycans to the CysR and Sp domains (19,20) might also help to facilitate localization of TS5 to the extracellular matrix.…”

Section: Discussionmentioning

confidence: 99%

“…The results of studies in HTB-94 chondrosarcoma cells suggest that the CysR domain helps localize TS5 in the extracellular matrix (19), whereas in TS4, the spacer domain is thought to mediate matrix binding (26 -29). Hyaluronan binding to the Dis domain (20) and binding of heparan sulfate proteoglycans to the CysR * This work was supported by the National Health & Medical Research Council and Sp domains, and possibly TS1 domain (19,20) might also help to facilitate matrix localization.…”

mentioning

confidence: 99%

A Disintegrin and Metalloproteinase with Thrombospondin Motifs-5 (ADAMTS-5) Forms Catalytically Active Oligomers

Kosasih

Last

Rogerson

et al. 2016

Journal of Biological Chemistry

View full text Add to dashboard Cite

The metalloproteinase ADAMTS-5 (A disintegrin and metalloproteinase with thrombospondin motifs) degrades aggrecan, a proteoglycan essential for cartilage structure and function. ADAMTS-5 is the major aggrecanase in mouse cartilage, and is also likely to be the major aggrecanase in humans. ADAMTS-5 is a multidomain enzyme, but the function of the C-terminal ancillary domains is poorly understood. We show that mutant ADAMTS-5 lacking the catalytic domain, but with a full suite of ancillary domains inhibits wild type ADAMTS activity, in vitro and in vivo, in a dominant-negative manner. The data suggest that mutant ADAMTS-5 binds to wild type ADAMTS-5; thus we tested the hypothesis that ADAMTS-5 associates to form oligomers. Co-elution, competition, and in situ PLA experiments using full-length and truncated recombinant ADAMTS-5 confirmed that ADAMTS-5 molecules interact, and showed that the catalytic and disintegrin-like domains support these intermolecular interactions. Cross-linking experiments revealed that recombinantADAMTS-5formedlarge,reduction-sensitiveoligomers with a nominal molecular mass of ϳ400 kDa. The oligomers were unimolecular and proteolytically active. ADAMTS-5 truncates comprising the disintegrin and/or catalytic domains were able to competitively block full-length ADAMTS-5-mediated aggrecan cleavage, measured by production of the G1-EGE 373 neoepitope. These results show that ADAMTS-5 oligomerization is required for full aggrecanase activity, and they provide evidence that blocking oligomerization inhibits ADAMTS-5 activity. The data identify the surface provided by the catalytic and disintegrin-like domains of ADAMTS-5 as a legitimate target for the design of aggrecanase inhibitors.ADAMTS-5 (TS5) is one member of a family of secreted, multidomain, zinc-dependent proteinases known as A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS).2 TS5 is a major aggrecan-degrading enzyme (aggrecanase) (1) mediating proteolysis and loss of aggrecan from articular cartilage in arthritic diseases. TS5 inhibitors are currently in development for arthritis therapies (2, 3), although ADAMTS-4 (TS4) might also have a role in human joint disease (4 -6). LGHG (7, 8). Fragments arising from proteolysis at these sites have been detected in cartilage and synovial fluids from patients with arthritic diseases including rheumatoid arthritis, osteoarthritis, and juvenile idiopathic arthritis (9 -12).TS5 is a multidomain enzyme comprising an N-terminal pro (Pro)-domain, a catalytic (Cat) domain, a disintegrin (Dis)-like domain, two thrombospondin (TS)-type 1 motifs, a cysteinerich (CysR) region, and a spacer (Sp) domain as reviewed previously (13-15). It is expressed as a zymogen that requires cleavage by pro-protein convertases (16, 17) at a polyarginine consensus sequence to remove the pro-domain. Further processing at the C terminus by autocatalysis gives rise to multiple forms of TS5, and recombinant C-terminal truncates have altered enzyme activity and substrate specificity (18,19). The active and ...

show abstract

Modulation of lubricin biosynthesis and tissue surface properties following cartilage mechanical injury

Jones¹,

Chen²,

Chai³

et al. 2008

Arthritis & Rheumatism

View full text Add to dashboard Cite

Objective. To evaluate the effects of injurious compression on the biosynthesis of lubricin at different depths within articular cartilage and to examine alterations in structure and function of the articular surface following mechanical injury.Methods. Bovine cartilage explants were subdivided into level 1, with intact articular surface, and level 2, containing middle and deep zone cartilage. Following mechanical injury, lubricin messenger RNA (mRNA) levels were monitored by quantitative reverse transcriptase-polymerase chain reaction, and soluble or cartilage-associated lubricin protein was analyzed by Western blotting and immunohistochemistry. Cartilage morphology was assessed by histologic staining, and tissue functionality was assessed by friction testing.Results. Two days after injury, lubricin mRNA expression was up-regulated ϳ3-fold for level 1 explants and was down-regulated for level 2 explants. Lubricin expression in level 1 cartilage returned to control levels after 6 days in culture. Similarly, lubricin protein synthesis and secretion increased in response to injury for level 1 explants and decreased for level 2 cartilage. Histologic staining revealed changes in the articular surface of level 1 explants following injury, with respect to glycosaminoglycan and collagen content. Injured level 1 explants displayed an increased coefficient of friction relative to controls. Conclusion. Our findings indicate that increased lubricin biosynthesis appears to be an early transient response of surface-layer cartilage to injurious compression. However, distinct morphologic changes occur with injury that appear to compromise the frictional properties of the tissue.Osteoarthritis (OA) is characterized by the degeneration of articular cartilage, leading to matrix fibrillation, fissuring, and the development of lesions. In the final stages of the disease, erosion of cartilage leads to painful bone-on-bone contact. The etiology of OA is complex and involves multiple biochemical, biomechanical, and genetic factors in addition to aging (1-3). Cartilage injury in young individuals is a prominent predisposing factor leading to increased risk of the subsequent development of OA (4,5) and, as such, represents a discrete pathologic event. Damage to the meniscus or ligaments sustained during traumatic joint injury causes instability, subjecting articular cartilage to abnormal biomechanical forces and resulting in the release of mediators of inflammation (6). Several animal models of OA are thus based on the observation that joint instability, i.e., via anterior cruciate ligament transaction or perturbation of the meniscus (7), results in the rapid onset of articular cartilage degeneration with an OA-like phenotype. The initial events following joint injury are thought to be crucial, since surgical interventions to restore joint stability do not seem to reduce the risk of developing posttraumatic OA (8).

show abstract

Aggrecanolysis in human osteoarthritis: confocal localization and biochemical characterization of ADAMTS5–hyaluronan complexes in articular cartilages

Cited by 101 publications

References 67 publications

Adamts5 Deletion Blocks Murine Dermal Repair through CD44-mediated Aggrecan Accumulation and Modulation of Transforming Growth Factor β1 (TGFβ1) Signaling

Adamts5 Deletion Blocks Murine Dermal Repair through CD44-mediated Aggrecan Accumulation and Modulation of Transforming Growth Factor β1 (TGFβ1) Signaling

A Disintegrin and Metalloproteinase with Thrombospondin Motifs-5 (ADAMTS-5) Forms Catalytically Active Oligomers

Modulation of lubricin biosynthesis and tissue surface properties following cartilage mechanical injury

Contact Info

Product

Resources

About