Bay-Jensen, Anne-C

Christensen, Britt

Karsdal, Morten

Kayed, Ashref

Kjelgaard-Petersen, Cecilie Freja

Sharma, Neha

Thudium, Christian S.

Background/Purpose:Several inflammatory cytokines and intracellular signaling pathways have been targeted in drug development with varying clinical results. Improved understanding of the intracellular signaling’s modulation of the extracellular matrix turnover could aid in selecting novel anti-inflammatory treatments for inflammatory arthritis. The aim of this study was to investigate the effect of small molecule inhibitors targeting 4 main pro-inflammatory signaling pathways (p38, Syk, IκBα, and STAT) on Oncostatin M (OSM) and Tumor Necrosis Factor α (TNFα) stimulated cartilage

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Downloaded from orbit.dtu.dk on: Dec 20, 2017Modulation of Cartilage Degradation Biomarkers Reflect the Activation and Inhibitionof Pro-Inflammatory Cytokine Signaling in an Ex Vivo Model of Bovine CartilageKjelgaard-Petersen, Cecilie Freja; Sharma, Neha; Kayed, Ashref; Christensen, Britt; Karsdal, Morten;Bay-Jensen, Anne-C; Thudium, Christian S.Published in:Arthritis and RheumatologyPublication date:2017Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Kjelgaard-Petersen, C. F., Sharma, N., Kayed, A., Christensen, B., Karsdal, M., Bay-Jensen, A-C., & Thudium,C. S. (2017). Modulation of Cartilage Degradation Biomarkers Reflect the Activation and Inhibition of Pro-Inflammatory Cytokine Signaling in an Ex Vivo Model of Bovine Cartilage. Arthritis and Rheumatology, 69(Suppl.10), [969].pdfcrowd.comDate: Monday, November 6, 2017Session Title: Cytokines, Mediators, Cell-Cell Adhesion, Cell Trafficking andAngiogenesis Poster II: Mesenchymal CellsDo React - But How?Session Type: ACR Poster Session BSession Time: 9:00AM-11:00AMABSTRACT NUMBER: 969Modulation of Cartilage Degradation Biomarkers Reflect theActivation and Inhibition of Pro-Inflammatory CytokineSignaling in an Ex Vivo Model of Bovine CartilageCecilie F. Kjelgaard-Petersen , Neha Sharma , Ashref Kayed , Britt Christensen , MortenKarsdal , Anne-C. Bay-Jensen  and Christian S. Thudium , Biomarkers and Research, NordicBioscience, Herlev, Denmark, Bioengineering, Technical University of Denmark, Kgs. Lyngby,Denmark, Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark, Research andBiomarkers, Nordic Bioscience, Herlev, Denmark, Biomolecular Sciences, University ofCopenhagen, Copenhagen, Denmark, Biomarkers and Reseacrh, Nordic Bioscience, Herlev,Denmark, Biomarkers and Reseach, Nordic Bioscience, Herlev, DenmarkMeeting: 2017 ACR/ARHP Annual MeetingDate of first publication: September 18, 2017Keywords: Biomarkers, cartilage, cytokines and signal transductionSESSION INFORMATIONBackground/Purpose:Several inflammatory cytokines and intracellular signaling pathways have been targeted in drugdevelopment with varying clinical results. Improved understanding of the intracellular signaling’smodulation of the extracellular matrix turnover could aid in selecting novel anti-inflammatorytreatments for inflammatory arthritis. The aim of this study was to investigate the effect of smallmolecule inhibitors targeting 4 main pro-inflammatory signaling pathways (p38, Syk, IκBα, andSTAT) on Oncostatin M (OSM) and Tumor Necrosis Factor α (TNFα) stimulated cartilage.  Methods:Full depth cartilage explants (FDC) were isolated from bovine knees. The FDCs were cultured for21 days without (WO) treatment, OSM [10ng/mL]+TNFα [2ng/mL] or OSM+TNFα together withSB203580, R406, TPCA-1 or Tofacitinib (Tofa) at 3 µM, 1 µM, 0.3 µM, or 0.1µM. DMSO was includedin WO and OSM+TNFα. The inhibitors were given at start of the experiment (preventive) or after10 days of OSM+TNFα stimuli (interventive). Activation of the p38, JNK, ERK, IκBα, and STAT3signaling pathways were assessed at time 0, 15’, 30’, 1h, 2h, 4h, 8h, and 24h of OSM+TNFα stimuliby western blot. The effect of the OSM+TNFα induced cartilage turnover was assessed by thebiomarkers AGNx1, C2M, and FFGV by ELISA in the conditioned medium.  Results:1,2 1,3 4,5 16 7 1 123 4567pdfcrowd.comWestern blot verified activation of p38, JNK, ERK, IκBα, and STAT3 signaling within 24h ofOSM+TNFα stimuli. OSM+TNFα significantly increased the biomarkers AGNx1 (P<0.001), FFGV(P<0.001) (Fig. 1), and C2M (P<0.001). Preventive SB203580 treatment had no effect on OSM+TNFαinduced AGNx1 release, while R406 (3µM: P=0.034), TPCA-1 (3µM: P<0.001) and Tofa (3µM: P<0.001)significantly decreased AGNx1 in a dose-dependent manner. All inhibitors given preventiveinhibited the release of C2M and FFGV (Fig.1a-d). SB203508 in a dose-dependent manner (3µM:P=0.001), while all concentrations of R406, TPCA-1, and Tofa significantly inhibited the release(P<0.05). Interventive treatment with SB203580 tended to decrease C2M and FFGV (Fig. 1e) release4 days after addition. R406, TPCA-1, and Tofa had no effect on C2M or FFGV after 4 days ofinterventive treatment, but tended to decrease C2M and significantly decreased FFGV (Fig. 1f-h)release after seven days of treatment in a dose-dependent manner (C2M: 3 µM, P≤0.055, FFGV).  Conclusion:In summary, we verified that OSM+TNFα stimulation activates a series of signaling pathways inour FDC culture. Using small molecule inhibitors targeting these individual pathways we foundthat they modulate the release of extracellular matrix degradation fragments in a spatial andtemporal manner. This is both dependent on the target signaling pathway and whether thetreatment is preventive or interventive.pdfcrowd.comDisclosure: C. F. Kjelgaard-Petersen, None; N. Sharma, None; A. Kayed, None; B. Christensen,Nordic Bioscience A/S, 3; M. Karsdal, Nordic Bioscience Diagnostic, 1,Symic Bio, 1,NordicBioscience A/S, 3; A. C. Bay-Jensen, Nordic Bioscience A/S, 1,Nordic Bioscience A/S, 3; C. S.Thudium, Nordic Bioscience Diagnostic, 3.To cite this abstract in AMA style:Kjelgaard-Petersen CF, Sharma N, Kayed A, Christensen B, Karsdal M, Bay-Jensen AC, ThudiumCS. Modulation of Cartilage Degradation Biomarkers Reflect the Activation and Inhibition ofPro-Inflammatory Cytokine Signaling in an Ex Vivo Model of Bovine Cartilage [abstract].Arthritis Rheumatol. 2017; 69 (suppl 10). http://acrabstracts.org/abstract/modulation-of-cartilage-degradation-biomarkers-reflect-the-activation-and-inhibition-of-pro-inflammatory-cytokine-signaling-in-an-ex-vivo-model-of-bovine-cartilage/. Accessed November 8, 2017.ACR Meeting Abstracts - http://acrabstracts.org/abstract/modulation-of-cartilage-degradation-biomarkers-reflect-the-activation-and-inhibition-of-pro-inflammatory-cytokine-signaling-in-an-ex-vivo-model-of-bovine-cartilage/

Modulation of Cartilage Degradation Biomarkers Reflect the Activation and Inhibition of Pro-Inflammatory Cytokine Signaling in an Ex Vivo Model of Bovine Cartilage

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Modulation of Cartilage Degradation Biomarkers Reflect the Activation and Inhibition of Pro-Inflammatory Cytokine Signaling in an Ex Vivo Model of Bovine Cartilage

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