Osteoarthritis and Cartilage
Volume 18, Issue 9 , Pages 1159-1166 , September 2010

A short-term pharmacodynamic model for monitoring aggrecanase activity: injection of monosodium iodoacetate (MIA) in rats and assessment of aggrecan neoepitope release in synovial fluid using novel ELISAs

  • C.A. Swearingen

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • These authors contributed equally to this work.
    • ,
  • M.G. Chambers

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • These authors contributed equally to this work.
    • ,
  • C. Lin

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • J. Marimuthu

      Affiliations

    • Discovery Chemistry, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • C.J. Rito

      Affiliations

    • Discovery Chemistry, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • Q.L. Carter

      Affiliations

    • Integrative Biology, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • J. Dotzlaf

      Affiliations

    • Integrative Biology, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • C. Liu

      Affiliations

    • Discovery Chemistry, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • S. Chandrasekhar

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • K.L. Duffin

      Affiliations

    • Integrative Biology, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • P.G. Mitchell

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • T.B. Durham

      Affiliations

    • Discovery Chemistry, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • M.R. Wiley

      Affiliations

    • Discovery Chemistry, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • ,
  • K. Thirunavukkarasu

      Affiliations

    • Musculoskeletal Research, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285, USA
    • Corresponding Author InformationAddress correspondence and reprint requests to: Kannan Thirunavukkarasu, Drop Code 0403, Lilly Research Labs, Eli Lilly and Company, Indianapolis, IN 46285.

Received 12 October 2009 ,Accepted 6 February 2010.

References 

  1. Burton-Wurster N, Todhunter RJ, Lust G. Animal models of osteoarthritis. In:  Woessner JF,  Howell DS editor. Joint Cartilage Degradation. 1993;p. 347–384
  2. Mankin HJ, Lippiello L. Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. J Bone Joint Surg Am. 1970;52:424–434
  3. Song RH, Tortorella MD, Malfait AM, Alston JT, Yang Z, Arner EC, et al. Aggrecan degradation in human articular cartilage explants is mediated by both ADAMTS-4 and ADAMTS-5. Arthritis Rheum. 2007;56:575–585
  4. Glasson S, Askew R, Sheppard B, Carito B, Blanchet T, Ma H, et al. Deletion of active ADAMTS5 prevents cartilage degradation in a murine model of osteoarthritis. Nature. 2005;434:644–648
  5. Glasson SS, Askew R, Sheppard B, Carito BA, Blanchet T, Ma HL, et al. Characterization of and osteoarthritis susceptibility in ADAMTS-4-knockout mice. Arthritis Rheum. 2004;50:2547–2558
  6. Majumdar MK, Askew R, Schelling S, Stedman N, Blanchet T, Hopkins B, et al. Double-knockout of ADAMTS-4 and ADAMTS-5 in mice results in physiologically normal animals and prevents the progression of osteoarthritis. Arthritis Rheum. 2007;56:3670–3674
  7. Stanton H, Rogerson F, East C, Golub S, Lawlor K, Meeker C, et al. ADAMTS5 is the major aggrecanase in mouse cartilage in vivo and in vitro. Nature. 2005;434:648–652
  8. Malfait AM, Liu RQ, Ijiri K, Komiya S, Tortorella MD. Inhibition of ADAM-TS4 and ADAM-TS5 prevents aggrecan degradation in osteoarthritic cartilage. J Biol Chem. 2002;277:22201–22208
  9. Sandy JD, Thompson V, Doege K, Verscharen C. The intermediates of aggrecanase-dependent cleavage of aggrecan in rat chondrosarcoma cells treated with interleukin-1. Biochem J. 2000;351(Pt 1):161–166
  10. Tortorella MD, Pratta M, Liu RQ, Austin J, Ross OH, Abbaszade I, et al. Sites of aggrecan cleavage by recombinant human aggrecanase-1 (ADAMTS-4). J Biol Chem. 2000;275(24):18566–18573
  11. Lohmander LS, Neame PJ, Sandy JD. The structure of aggrecan fragments in human synovial fluid. Evidence that aggrecanase mediates cartilage degradation in inflammatory joint disease, joint injury, and osteoarthritis. Arthritis Rheum. 1993;36:1214–1222
  12. Ameye LG, Young MF. Animal models of osteoarthritis: lessons learned while seeking the “Holy Grail”. Curr Opin Rheumatol. 2006;18:537–547
  13. Bendele AM. Animal models of osteoarthritis. J Musculoskelet Neuronal Interact. 2001;1:363–376
  14. Malfait AM, Tortorella M, Thompson J, Hills R, Meyer DM, Jaffee BD, et al. Intra-articular injection of tumor necrosis factor-a in the rat: an acute and reversible in vivo model of cartilage proteoglycan degradation. Osteoarthritis Cartilage. 2009;17:627–635
  15. Janusz MJ, Little CB, King LE, Hookfin EB, Brown KK, Heitmeyer SA, et al. Detection of aggrecanase- and MMP-generated catabolic neoepitopes in the rat iodoacetate model of cartilage degeneration. Osteoarthritis Cartilage. 2004;12:720–728
  16. Pratta MA, Su JL, Leesnitzer MA, Struglics A, Larsson S, Lohmander LS, et al. Development and characterization of a highly specific and sensitive sandwich ELISA for detection of aggrecanase-generated aggrecan fragments. Osteoarthritis Cartilage. 2006;14:702–713
  17. Hughes CE, Caterson B, Fosang AJ, Roughley PJ, Mort JS. Monoclonal antibodies that specifically recognize neoepitope sequences generated by ‘aggrecanase’ and matrix metalloproteinase cleavage of aggrecan: application to catabolism in situ and in vitro. Biochem J. 1995;305(Pt 3):799–804
  18. Larsson S, Lohmander LS, Struglics A. Synovial fluid level of aggrecan ARGS fragments is a more sensitive marker of joint disease than glycosaminoglycan or aggrecan levels: a cross-sectional study. Arthritis Res Ther. 2009;11:R92
  19. Oegema TR, Hascall VC, Dziewiatkowski DD. Isolation and characterization of proteoglycans from the Swarm rat chondrosarcoma. J Biol Chem. 1975;250:6151
  20. Plaas AHK, Barry FP, Wong-Palms S. Keratan sulfate substitution on cartilage matrix molecules. In:  Kuettner KE,  Schleyerbach R,  Peyron JG,  Hascall VC editor. Articular Cartilage and Osteoarthritis. New York: Raven Press; 1992;p. 69
  21. Venn G, Mason RM. Absence of keratan sulphate from skeletal tissues of mouse and rat. Biochem J. 1985;228:443–450
  22. Sandy JD, Flannery CR, Neame PJ, Lohmander LS. The structure of aggrecan fragments in human synovial fluid. Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. J Clin Invest. 1992;89:1512–1516
  23. Mercuri FA, Doege KJ, Arner EC, Pratta MA, Last K, Fosang AJ. Recombinant human aggrecan G1-G2 exhibits native binding properties and substrate specificity for matrix metalloproteinases and aggrecanase. J Biol Chem. 1999;274(45):32387–32395
  24. Perkins SJ, Nealis AS, Dudhia J, Hardingham TE. Immunoglobulin fold and tandem repeat structures in proteoglycan N-terminal domains and link protein. J Mol Biol. 1989;206:737–753
  25. Carter QL, Dotzlaf J, Swearingen C, Brittain I, Chambers M, Duffin K, et al. Development and characterization of a novel ELISA based assay for the quantitation of sub-nanomolar levels of neoepitope exposed NITEGE-containing aggrecan fragments. J Immunol Methods. 2007;328:162–168
  26. Tortorella MD, Liu RQ, Burn T, Newton RC, Arner E. Characterization of human aggrecanase 2 (ADAM-TS5): substrate specificity studies and comparison with aggrecanase 1 (ADAM-TS4). Matrix Biol. 2002;21(6):499–511
  27. Knight CG, Willenbrock F, Murphy G. A novel coumarin-labelled peptide for sensitive continuous assays of the matrix metalloproteinases. FEBS Lett. 1992;296:263–266
  28. Swearingen C, Thirunavukkarasu K. Development of an alphascreen assay to identify inhibitors of aggrecanase activity. Trans Orthop Res Soc. 2007;(Abstract # 0085)
  29. Mitchell PG, Yocum SA, Lopresti-Morrow LL, Tobiassen LM, Bliven M, Otterness I, et al. Characterization of the cartilage protective activity of a dual inhibitor of matrix metalloproteinases and ADAMTS-4. Trans Orthop Res Soc. 2003;(Abstract # 703)
  30. Noe MC, Natarajan V, Snow SL, Wolf-Gouveia LA, Mitchell PG, Lopresti-Morrow L, et al. Discovery of 3-OH-3-methylpipecolic hydroxamates: potent orally active inhibitors of aggrecanase and MMP-13. Bioorg Med Chem Lett. 2005;15:3385–3388
  31. Lovejoy B, Welch AR, Carr S, Luong C, Broka C, Hendricks RT, et al. Crystal structures of MMP-1 and -13 reveal the structural basis for selectivity of collagenase inhibitors. Nat Struct Biol. 1999;6:217–221
  32. Pratta MA, Yao W, Decicco C, Tortorella MD, Liu R-Q, Copeland RA, et al. Aggrecan protects cartilage collagen from proteolytic cleavage. J Biol Chem. 2003;278:45539–45545
  33. Kalbhen DA. Chemical model of osteoarthritis – pharmacological evaluation. Rheumatology. 1987;14:130–131
  34. Van der Kraan PM, Vitters EL, Van de Putte LBA, Van den Berg WB. Development of osteoarthritic lesions in mice by ‘Metabolic’ and ‘Mechanical’ alterations in the knee joints. Am J Pathol. 1989;35:1001–1014
  35. Bove SE, Calcaterra SL, Brooker RM, Huber CM, Guzman RE, Juneau PL, et al. Weight bearing as a measure of disease progression and efficacy of anti-inflammatory compounds in a model of monosodium iodoacetate-induced osteoarthritis. Osteoarthritis Cartilage. 2003;11:821–830
  36. Janusz MJ, Hookfin EB, Heitmeyer SA, Woessner JF, Freemont AJ, Hoyland JA, et al. Moderation of iodoacetate-induced experimental osteoarthritis in rats by matrix metalloproteinase inhibitors. Osteoarthritis Cartilage. 2001;9:751–760
  37. Flannery CR, Lark MW, Sandy JD. Identification of a stromelysin cleavage site within the interglobular domain of human aggrecan. Evidence for proteolysis at this site in vivo in human articular cartilage. J Biol Chem. 1992;267:1008–1014
  38. Fosang AJ, Last K, Knäuper V, Neame PJ, Murphy G, Hardingham TE, et al. Fibroblast and neutrophil collagenases cleave at two sites in the cartilage aggrecan interglobular domain. Biochem J. 1993;295(Pt 1):273–276
  39. Fosang AJ, Neame PJ, Last K, Hardingham TE, Murphy G, Hamilton JA. The interglobular domain of cartilage aggrecan is cleaved by PUMP, gelatinases, and cathepsin B. J Biol Chem. 1992;267:19470–19474
  40. Chockalingam PS, Zeng W, Morris EA, Flannery CR. Release of hyaluronan and hyaladherins (aggrecan G1 domain and link proteins) from articular cartilage exposed to ADAMTS-4 (aggrecanase 1) or ADAMTS-5 (aggrecanase 2). Arthritis Rheum. 2004;50:2839–2848
  41. Fosang AJ, Last K, Maciewicz RA. Aggrecan is degraded by matrix metalloproteinases in human arthritis. Evidence that matrix metalloproteinase and aggrecanase activities can be independent. J Clin Invest. 1996;98(10):2292–2299
  42. Sandy JD, Verscharen C. Analysis of aggrecan in human knee cartilage and synovial fluid indicates that aggrecanase (ADAMTS) activity is responsible for the catabolic turnover and loss of whole aggrecan whereas other protease activity is required for C-terminal processing in vivo. Biochem J. 2001;358(Pt 3):615–626
  43. Flannery CR, Little CB, Caterson B. Molecular cloning and sequence analysis of the aggrecan interglobular domain from porcine, equine, bovine and ovine cartilage: comparison of proteinase-susceptible regions and sites of keratan sulfate substitution. Matrix Biol. 1998;16:507–511

PII: S1063-4584(10)00215-3

doi: 10.1016/j.joca.2010.02.019

Osteoarthritis and Cartilage
Volume 18, Issue 9 , Pages 1159-1166 , September 2010

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