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Investigating the Robustness and Diagnostic Potential of Extracellular Matrix Remodelling Biomarkers in Alkaptonuria

  • F. GenoveseEmail author
  • A. S. Siebuhr
  • K. Musa
  • J. A. Gallagher
  • A. M. Milan
  • M. A. Karsdal
  • J. Rovensky
  • A. C. Bay-Jensen
  • L. R. Ranganath
Research Report
Part of the JIMD Reports book series (JIMD, volume 24)

Abstract

Background and aim: Alkaptonuria (AKU) clinical manifestations resemble severe arthritis. The Suitability of Nitisinone in Alkaptonuria 1 (SONIA 1) study is a dose-finding trial for nitisinone treatment of AKU patients. We tested a panel of serum and urinary biomarkers reflecting extracellular matrix remodelling (ECMR) of cartilage, bone and connective tissue in SONIA 1 patients to identify non-invasive and diagnostic biomarkers of tissue turnover in AKU.

Methods: Fasted serum and urine were retrieved from 40 SONIA 1 patients and 44 healthy controls. Established biomarkers of bone remodelling (CTX-I, P1NP, OC), cartilage remodelling (CTX-II, C2M, AGNx1) and inflammation (CRPM) as well as exploratory biomarkers of ECMR (C6M, VCANM, MIM, TIM) were measured at baseline in serum and urine by means of enzyme-linked immunosorbent assays (ELISAs) or automated systems (Elecsys 2010).

Results: The levels of bone resorption (CTX-I) and cartilage degradation (C2M) were elevated in AKU patients as compared to controls (p > 0.0001 and p = 0.03, respectively). Also tissue inflammation (CRPM) was elevated in AKU patients (p = 0.01). In addition all four exploratory biomarkers of ECMR (C6M, VCANM, MIM, TIM) were elevated in AKU patients compared to healthy controls. CTX-II was the only biomarker to be reduced in AKU patients. TIM was the only marker that showed a higher concentration than the normal assay range in AKU patients.

Conclusions: We have identified new potential biomarkers for assessment of cartilage, bone and cardiovascular remodelling in AKU and demonstrated the robustness of the assays used to measure the biomarker concentration in biological fluids.

Keywords

Urine Creatinine Cartilage Degradation Extracellular Matrix Remodelling Homogentisic Acid Cardiovascular Remodelling 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgements

This study was funded by the European Union Seventh Framework Programme (project 304985) and by the Danish Research Fund (“Den Danske Forskningsfond”).

Supplementary material

346908_1_En_430_MOESM1_ESM.zip (60 kb)
(DOC 23 kb)
346908_1_En_430_MOESM2_ESM.zip (48 kb)
(DOC 89 kb)

References

  1. Arends S, Spoorenberg A, Houtman PM, Leijsma MK, Bos R, Kallenberg CG, Groen H, Brouwer E, van der V (2012) The effect of three years of TNFalpha blocking therapy on markers of bone turnover and their predictive value for treatment discontinuation in patients with ankylosing spondylitis: a prospective longitudinal observational cohort study. Arthritis Res Ther 14:R98PubMedCentralCrossRefPubMedGoogle Scholar
  2. Barascuk N, Vassiliadis E, Zheng Q, Wang Y, Wang W, Larsen L, Rasmussen LM, Karsdal MA (2011) Levels of circulating MMCN-151, a degradation product of mimecan, reflect pathological extracellular matrix remodeling in apolipoprotein E knockout mice. Biomark Insights 6:97–106PubMedCentralCrossRefPubMedGoogle Scholar
  3. Barascuk N, Genovese F, Larsen L, Byrjalsen I, Zheng Q, Sun S, Hosbond S, Poulsen TS, Diederichsen A, Jensen JM, Mickley H, Register TC, Rasmussen LM, Leeming DJ, Christiansen C, Karsdal MA (2013) A MMP derived versican neo-epitope is elevated in plasma from patients with atherosclerotic heart disease. Int J Clin Exp Med 6:174–184PubMedCentralPubMedGoogle Scholar
  4. Bay-Jensen AC, Liu Q, Byrjalsen I, Li Y, Wang J, Pedersen C, Leeming DJ, Dam EB, Zheng Q, Qvist P, Karsdal MA (2011) Enzyme-linked immunosorbent assay (ELISAs) for metalloproteinase derived type II collagen neoepitope, CIIM–increased serum CIIM in subjects with severe radiographic osteoarthritis. Clin Biochem 44:423–429CrossRefPubMedGoogle Scholar
  5. Bay-Jensen AC, Platt A, Byrjalsen I, Vergnoud P, Christiansen C, Karsdal MA (2014) Effect of tocilizumab combined with methotrexate on circulating biomarkers of synovium, cartilage, and bone in the LITHE study. Semin Arthritis Rheum 43:470–478CrossRefPubMedGoogle Scholar
  6. Curtis SL, Roberts NB, Ranganath LR (2014) Interferences of homogentisic acid (HGA) on routine clinical chemistry assays in serum and urine and the implications for biochemical monitoring of patients with alkaptonuria. Clin Biochem 47:640–647CrossRefPubMedGoogle Scholar
  7. Dam EB, Byrjalsen I, Karsdal MA, Qvist P, Christiansen C (2009a) Increased urinary excretion of C-telopeptides of type II collagen (CTX-II) predicts cartilage loss over 21 months by MRI. Osteoarthritis Cartilage 17:384–389CrossRefPubMedGoogle Scholar
  8. Dam EB, Loog M, Christiansen C, Byrjalsen I, Folkesson J, Nielsen M, Qazi AA, Pettersen PC, Garnero P, Karsdal MA (2009b) Identification of progressors in osteoarthritis by combining biochemical and MRI-based markers. Arthritis Res Ther 11:R115PubMedCentralCrossRefPubMedGoogle Scholar
  9. Dorleijn DM, Luijsterburg PA, Bay-Jensen AC, Siebuhr AS, Karsdal MA, Rozendaal RM, Bos PK, Bierma-Zeinstra SM (2014) Association between biochemical cartilage markers and clinical symptoms in patients with hip osteoarthritis: cohort study with 2-year follow-up. Osteoarthritis Cartilage 23:57–62CrossRefPubMedGoogle Scholar
  10. Garnero P, Ferreras M, Karsdal MA, Nicamhlaoibh R, Risteli J, Borel O, Qvist P, Delmas PD, Foged NT, Delaisse JM (2003) The type I collagen fragments ICTP and CTX reveal distinct enzymatic pathways of bone collagen degradation. J Bone Miner Res 18:859–867CrossRefPubMedGoogle Scholar
  11. Hashimoto J, Garnero P, van der HD, Miyasaka N, Yamamoto K, Kawai S, Takeuchi T, Yoshikawa H, Nishimoto N (2009) A combination of biochemical markers of cartilage and bone turnover, radiographic damage and body mass index to predict the progression of joint destruction in patients with rheumatoid arthritis treated with disease-modifying anti-rheumatic drugs. Mod Rheumatol 19:273–282Google Scholar
  12. Karsdal MA, Byrjalsen I, Riis BJ, Christiansen C (2008) Investigation of the diurnal variation in bone resorption for optimal drug delivery and efficacy in osteoporosis with oral calcitonin. BMC Clin Pharmacol 8:12PubMedCentralCrossRefPubMedGoogle Scholar
  13. Karsdal MA, Byrjalsen I, Bay-Jensen AC, Henriksen K, Riis BJ, Christiansen C (2010) Biochemical markers identify influences on bone and cartilage degradation in osteoarthritis–the effect of sex, Kellgren-Lawrence (KL) score, body mass index (BMI), oral salmon calcitonin (sCT) treatment and diurnal variation. BMC Musculoskelet Disord 11:125PubMedCentralCrossRefPubMedGoogle Scholar
  14. Kenny D, Ptacin MJ, Bamrah VS, Almagro U (1990) Cardiovascular ochronosis: a case report and review of the medical literature. Cardiology 77:477–483CrossRefPubMedGoogle Scholar
  15. Leeming DJ, Larsen DV, Zhang C, Hi Y, Veidal SS, Nielsen RH, Henriksen K, Zheng Q, Barkholt V, Riis BJ, Byrjalsen I, Qvist P, Karsdal MA (2010) Enzyme-linked immunosorbent serum assays (ELISAs) for rat and human N-terminal pro-peptide of collagen type I (PINP)–assessment of corresponding epitopes. Clin Biochem 43:1249–1256CrossRefPubMedGoogle Scholar
  16. Lok ZS, Goldstein J, Smith JA (2013) Alkaptonuria-associated aortic stenosis. J Card Surg 28:417–420CrossRefPubMedGoogle Scholar
  17. Millucci L, Ghezzi L, Paccagnini E, Giorgetti G, Viti C, Braconi D, Laschi M, Geminiani M, Soldani P, Lupetti P, Orlandini M, Benvenuti C, Perfetto F, Spreafico A, Bernardini G, Santucci A (2014a) Amyloidosis, inflammation, and oxidative stress in the heart of an alkaptonuric patient. Mediators Inflamm 2014:258471PubMedCentralCrossRefPubMedGoogle Scholar
  18. Millucci L, Ghezzi L, Braconi D, Laschi M, Geminiani M, Amato L, Orlandini M, Benvenuti C, Bernardini G, Santucci A (2014b) Secondary amyloidosis in an alkaptonuric aortic valve. Int J Cardiol 172:e121–e123PubMedCentralCrossRefPubMedGoogle Scholar
  19. Pedersen SJ, Sorensen IJ, Garnero P, Johansen JS, Madsen OR, Tvede N, Hansen MS, Thamsborg G, Andersen LS, Majgaard O, Loft AG, Erlendsson J, Asmussen K, Jurik AG, Moller J, Hasselquist M, Mikkelsen D, Skjodt T, Lambert R, Hansen A, Ostergaard M (2011) ASDAS, BASDAI and different treatment responses and their relation to biomarkers of inflammation, cartilage and bone turnover in patients with axial spondyloarthritis treated with TNFalpha inhibitors. Ann Rheum Dis 70:1375–1381CrossRefPubMedGoogle Scholar
  20. Preston AJ, Keenan CM, Sutherland H, Wilson PJ, Wlodarski B, Taylor AM, Williams DP, Ranganath LR, Gallagher JA, Jarvis JC (2014) Ochronotic osteoarthropathy in a mouse model of alkaptonuria, and its inhibition by nitisinone. Ann Rheum Dis 73:284–289CrossRefPubMedGoogle Scholar
  21. Ranganath LR, Jarvis JC, Gallagher JA (2013) Recent advances in management of alkaptonuria (invited review; best practice article). J Clin Pathol 66:367–373CrossRefPubMedGoogle Scholar
  22. Ranganath LR, Milan AM, Huges AT (2014) Suitability of nitisinone in alkaptonuria 1 (SONIA 1): an international, multicenter, randomized, open-label, no-treatment controlled, parallel-group, dose–response study to investigate the effect of once daily nitisinone on 24-hour urinary homogentisic acid excretion in patients with alkaptonuria after 4 weeks of treatment. Ann Rheum Dis 0:1–6Google Scholar
  23. Rosenquist C, Qvist P, Bjarnason N, Christiansen C (1995) Measurement of a more stable region of osteocalcin in serum by ELISA with two monoclonal antibodies. Clin Chem 41:1439–1445PubMedGoogle Scholar
  24. Siebuhr AS, Wang J, Karsdal M, Bay-Jensen AC, Jin Y, Zheng Q (2012) Matrix metalloproteinase-dependent turnover of cartilage, synovial membrane, and connective tissue is elevated in rats with collagen induced arthritis. J Transl Med 10:195Google Scholar
  25. Siebuhr AS, Petersen KK, Rendt-Nielsen Z, Egsgaard LL, Eskehave T, Christiansen C, Simonsen O, Hoeck HC, Karsdal MA, Bay-Jensen AC (2014) Identification and characterisation of osteoarthritis patients with inflammation derived tissue turnover. Osteoarthritis Cartilage 22:44–50Google Scholar
  26. Skjot-Arkil H, Schett G, Zhang C, Larsen DV, Wang Y, Zheng Q, Larsen MR, Nawrocki A, Bay-Jensen AC, Henriksen K, Christiansen C, Alexandersen P, Leeming DJ, Karsdal MA (2012) Investigation of two novel biochemical markers of inflammation, matrix metalloproteinase and cathepsin generated fragments of C-reactive protein, in patients with ankylosing spondylitis. Clin Exp Rheumatol 30:371–379PubMedGoogle Scholar
  27. Soder S, Hambach L, Lissner R, Kirchner T, Aigner T (2002) Ultrastructural localization of type VI collagen in normal adult and osteoarthritic human articular cartilage. Osteoarthritis Cartilage 10:464–470CrossRefPubMedGoogle Scholar
  28. Syversen SW, Goll GL, van der HD, Landewe R, Gaarder PI, Odegard S, Haavardsholm EA, Kvien TK (2009) Cartilage and bone biomarkers in rheumatoid arthritis: prediction of 10-year radiographic progression. J Rheumatol 36:266–272Google Scholar
  29. Taylor AM, Boyde A, Wilson PJ, Jarvis JC, Davidson JS, Hunt JA, Ranganath LR, Gallagher JA (2011) The role of calcified cartilage and subchondral bone in the initiation and progression of ochronotic arthropathy in alkaptonuria. Arthritis Rheum 63:3887–3896CrossRefPubMedGoogle Scholar
  30. Taylor AM, Boyde A, Davidson JS, Jarvis JC, Ranganath LR, Gallagher JA (2012a) Identification of trabecular excrescences, novel microanatomical structures, present in bone in osteoarthropathies. Eur Cell Mater 23:300–308PubMedGoogle Scholar
  31. Taylor AM, Preston AJ, Paulk NK, Sutherland H, Keenan CM, Wilson PJ, Wlodarski B, Grompe M, Ranganath LR, Gallagher JA, Jarvis JC (2012b) Ochronosis in a murine model of alkaptonuria is synonymous to that in the human condition. Osteoarthritis Cartilage 20:880–886PubMedCentralCrossRefPubMedGoogle Scholar
  32. Vassiliadis E, Rasmussen LM, Byrjalsen I, Larsen DV, Chaturvedi R, Hosbond S, Saabye L, Diederichsen AC, Genovese F, Duffin KL, Zheng Q, Chen X, Leeming DJ, Christiansen C, Karsdal MA (2012) Clinical evaluation of a matrix metalloproteinase-12 cleaved fragment of titin as a cardiovascular serological biomarker. J Transl Med 10:140PubMedCentralCrossRefPubMedGoogle Scholar
  33. Veidal SS, Karsdal MA, Vassiliadis E, Nawrocki A, Larsen MR, Nguyen QH, Hagglund P, Luo Y, Zheng Q, Vainer B, Leeming DJ (2011) MMP mediated degradation of type VI collagen is highly associated with liver fibrosis–identification and validation of a novel biochemical marker assay. PLoS One 6:e24753PubMedCentralCrossRefPubMedGoogle Scholar
  34. Vosse D, Landewe R, Garnero P, van der HD, van der LS, Geusens P (2008) Association of markers of bone- and cartilage-degradation with radiological changes at baseline and after 2 years follow-up in patients with ankylosing spondylitis. Rheumatology (Oxford) 47:1219–1222CrossRefGoogle Scholar
  35. Wang B, Chen P, Jensen AC, Karsdal MA, Madsen SH, Sondergaard BC, Zheng Q, Qvist P (2009) Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments. BMC Res Notes 2:259PubMedCentralCrossRefPubMedGoogle Scholar
  36. Zatkova A (2011) An update on molecular genetics of Alkaptonuria (AKU). J Inherit Metab Dis 34:1127–1136CrossRefPubMedGoogle Scholar

Copyright information

© SSIEM and Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • F. Genovese
    • 1
    Email author
  • A. S. Siebuhr
    • 1
  • K. Musa
    • 1
  • J. A. Gallagher
    • 2
  • A. M. Milan
    • 2
    • 3
  • M. A. Karsdal
    • 1
  • J. Rovensky
    • 4
  • A. C. Bay-Jensen
    • 1
  • L. R. Ranganath
    • 2
    • 3
  1. 1.Nordic BioscienceHerlevDenmark
  2. 2.Department of Musculoskeletal Biology, Institute of Ageing and Chronic DiseaseUniversity of LiverpoolLiverpoolUK
  3. 3.Department of Clinical Biochemistry and Metabolic MedicineRoyal Liverpool University Hospital TrustLiverpoolUK
  4. 4.National Institute of Rheumatic DiseasesPiešťanySlovakia

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