Novel Cartilage Oligomeric Matrix Protein (COMP) Neoepitopes Identified in Synovial Fluids from Patients with Joint Diseases Using Affinity Chromatography and Mass Spectrometry

Åhrman, Emma; Lorenzo, Pilar; Holmgren, Kristin; Grodzinsky, Alan J.; Dahlberg, Leif; Saxne, Tore; Önnerfjord, Patrik

doi:10.1074/jbc.m114.554683

Cited by 36 publications

(39 citation statements)

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“…Further, cells responding to IL-1a in our model may be endothelial cells and resident immune cells, notably macrophages. The process of matrix degradation in cartilage explants after cytokine treatment was studied previously with MS, and the focus most often has been on the components released from cartilage (35)(36)(37). Because we were interested in investigating how alterations in the cartilage surface affect how it interacts with complement, we focused on determining the protein content in the residual cartilage explant upon IL-1a stimulation.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Mass Spectrometry To Study Inflammatory Cartilage Degradation and Resulting Interactions with the Complement System

Fürst

Åhrman

Bratteby

et al. 2016

The Journal of Immunology

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Joint diseases are often characterized by inflammatory processes that result in pathological changes in joint tissues, including cartilage degradation and release of components into the synovial fluid. The complement system plays a central role in promoting the inflammation. Because several cartilage proteins are known to interact with complement, causing either activation or inhibition of the system, we aimed to investigate these interactions comprehensively. Bovine cartilage explants were cultured with IL-1α to induce cartilage degradation, followed by incubation with human serum. Label-free selected reaction monitoring mass spectrometry was used to specifically quantify complement proteins interacting with the cartilage explant. In parallel, the time-dependent degradation of cartilage was detected using mass spectrometry analysis (liquid chromatography–tandem mass spectrometry). Complement proteins resulting from activation of the classical, alternative, and terminal pathways were detected on IL-1α–stimulated cartilage at time points when clear alterations in extracellular matrix composition had occurred. Increased levels of the complement activation product C4d, as detected by ELISA in serum after incubation with IL-1α–stimulated cartilage, confirmed the selected reaction monitoring results indicating complement activation. Further, typical activated (cleaved) C3 fragments were detected by Western blotting in extracts of IL-1α–stimulated cartilage. No complement activation was triggered by cartilage cultured in the absence of IL-1α. Components released from IL-1α–stimulated cartilage during culture had an inhibitory effect on complement activation. These were released after a longer incubation period with IL-1α and may represent a feedback reaction to cartilage-triggered complement activation observed after a shorter incubation period.

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Section: Discussionmentioning

confidence: 99%

Quantitative Mass Spectrometry To Study Inflammatory Cartilage Degradation and Resulting Interactions with the Complement System

Fürst

Åhrman

Bratteby

et al. 2016

The Journal of Immunology

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“…The culture medium from such studies (involving treatment of explants with various inflammatory, injurious, or therapeutic factors) has been used as a surrogate for synovial fluid in order to reduce complexity and to identify the response of cartilage to specific treatments. Recent proteomics studies of the cartilage secretome in response to specific cytokines [15–20] have enabled the discovery of new macromolecules and novel cleavage products. This advantage is now further enhanced by obtaining proteomic data to track a broad array of matrix molecules and fragments secreted by explants following mechanical injury and cytokine co-culture.…”

Section: Introductionmentioning

confidence: 99%

Quantitative proteomics analysis of cartilage response to mechanical injury and cytokine treatment

Wang

Khabut

et al. 2017

Matrix Biology

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Mechanical damage at the time of joint injury and the ensuing inflammatory response associated with elevated levels of pro-inflammatory cytokines in the synovial fluid, are reported to contribute to the progression to osteoarthritis after injury. In this exploratory study, we used a targeted proteomics approach to follow the progression of matrix degradation in response to mechanical damage and cytokine treatment of human knee cartilage explants, and thereby to study potential molecular biomarkers. This proteomics approach allowed us to unambiguously identify and quantify multiple peptides and proteins in the cartilage medium and explants upon treatment with +/− injurious compression +/−cytokines, treatments that mimic the earliest events in post-traumatic OA. We followed degradation of different protein domains, e.g., G1/G2/G3 of aggrecan, by measuring representative peptides of matrix proteins released into the medium at 7 time points throughout the 21-day culture period. COMP neo epitopes, which were previously identified in the synovial fluid of knee injury/OA patients, were also released by these human cartilage explants treated with cyt and cyt+inj. The absence of collagen pro-peptides and elevated levels of specific COMP and COL3A1 neo-epitopes after human knee trauma may be relevant as potential biomarkers for post-traumatic OA. This model system thereby enables study of the kinetics of cartilage degradation and the identification of biomarkers within cartilage explants and those released to culture medium. Discovery proteomics revealed that candidate proteases were identified after specific treatment conditions, including MMP1, MMP-3, MMP-10 and MMP-13.

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“…Increased COMP has been associated with fibrogenesis in systemic sclerosis [18], skin keloids [19, 20], vascular atherosclerosis [21], lung fibrosis [22] and chronic pancreatitis [23] as well as with other conditions such as rheumatoid arthritis [24, 25], osteoarthritis [26, 27], pseudoachondroplasia [28], acute trauma [29] and systemic lupus erythematosus [30, 31]. Recently, COMP has been proposed as a novel non-invasive marker for assessing cirrhosis and the risk of hepatocellular carcinoma [32] and as a biomarker of liver fibrosis in chronic hepatitis C [33].…”

Section: Introductionmentioning

confidence: 99%

Cartilage oligomeric matrix protein participates in the pathogenesis of liver fibrosis

Magdaleno

Arriazu

Galarreta

et al. 2016

Journal of Hepatology

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Background & Aims Liver fibrosis is characterized by significant accumulation of extracellular matrix (ECM) proteins, mainly fibrillar collagen-I, as a result of persistent liver injury. Cartilage oligomeric matrix protein (COMP) is largely found in the ECM of skeletal tissue. Increased COMP expression has been associated with fibrogenesis in systemic sclerosis, lung fibrosis, chronic pancreatitis, cirrhosis and hepatocellular carcinoma. We hypothesized that COMP could induce fibrillar collagen-I deposition and participate in matrix remodeling thus contributing to the pathophysiology of liver fibrosis. Methods Thioacetamide (TAA) and carbon tetrachloride (CCl4) were used to induce liver fibrosis in wild-type (WT) and Comp−/− mice. In vitro experiments were performed with primary hepatic stellate cells (HSCs). Results COMP expression was detected in livers from control WT mice and was up-regulated in response to either TAA or CCl4-induced liver fibrosis. TAA-treated or CCl4-injected Comp−/− mice showed less liver injury, inflammation and fibrosis compared to their corresponding control WT mice. Challenge of HSCs with recombinant COMP (rCOMP) induced intra- plus extracellular collagen-I deposition and increased matrix metalloproteinases (MMPs) 2, 9 and 13, albeit similar expression of TGFβ protein in addition to Tgfβ, Tnfα and tissue inhibitor of metalloproteinases-1 (Timp1) mRNAs. We demonstrated that COMP binds collagen-I; yet, it does not prevent collagen-I cleavage by MMP1. Last, rCOMP induced collagen-I expression in HSCs via CD36 receptor signaling and activation of the MEK1/2-pERK1/2 pathway. Conclusion These results suggest that COMP contributes to liver fibrosis by regulating collagen-I deposition.

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Novel Cartilage Oligomeric Matrix Protein (COMP) Neoepitopes Identified in Synovial Fluids from Patients with Joint Diseases Using Affinity Chromatography and Mass Spectrometry

Cited by 36 publications

References 36 publications

Quantitative Mass Spectrometry To Study Inflammatory Cartilage Degradation and Resulting Interactions with the Complement System

Quantitative Mass Spectrometry To Study Inflammatory Cartilage Degradation and Resulting Interactions with the Complement System

Quantitative proteomics analysis of cartilage response to mechanical injury and cytokine treatment

Cartilage oligomeric matrix protein participates in the pathogenesis of liver fibrosis

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