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High calcium concentration and calcium oxalate crystals cause significant inaccuracies in the measurement of urinary osteopontin by enzyme linked immunosorbent assay

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Abstract

Strong evidence that osteopontin (OPN) is a determinant of urolithiasis has prompted studies comparing the protein’s urinary excretion in healthy subjects and stone formers. However, reported mean urinary values have varied widely, from <1 μg/mL to more than 20 times that value. Since OPN binds to CaOx crystals, the presence of crystals in urine may cause underestimation of its urinary levels. Using a commercial ELISA, we measured urinary OPN levels in the presence of endogenous or exogenous CaOx monohydrate (COM) and dihydrate (COD) crystals. OPN concentrations decreased in the presence of endogenous and exogenous CaOx crystals, but never below 2 μg/mL. Increasing the urinary calcium concentration decreased detectable OPN levels, possibly as a result of changes in the three-dimensional conformation of the protein. Because calcium concentration and the formation of CaOx crystals cannot be controlled in urine, the use of urinary OPN levels as a biomarker for any human pathology must be seriously questioned, but particularly for the investigation of stone formers in whom hypercalciuria and crystalluria are more common than in healthy subjects.

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References

  1. Denhardt DT, Guo X (1993) Osteopontin: a protein with diverse functions. FASEB J 7:1475–1482

    CAS  PubMed  Google Scholar 

  2. Oldberg A, Franzen A, Heinegard D (1986) Cloning and sequence analysis of rat bone sialoprotein (osteopontin) cDNA reveals an Arg-Gly-Asp cell-binding sequence. Proc Nat Acad Sci USA 83:8819–8823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Qin C, Baba O, Butler WT (2004) Post-translational modifications of SIBLING proteins and their roles in osteogenesis and dentinogenesis. Crit Rev Oral Biol Med 15:126–136

    Article  CAS  PubMed  Google Scholar 

  4. Fisher LW, Torchia DA, Fohr B, Young MF, Fedarko NS (2001) Flexible structures of SIBLING proteins, bone sialoprotein, and osteopontin. Biochem Biophys Res Commun 280:460–465

    Article  CAS  PubMed  Google Scholar 

  5. Hoyer JR, Otvos L Jr, Urge L (1995) Osteopontin in urinary stone formation. Ann NY Acad Sci 760:257–265

    Article  CAS  PubMed  Google Scholar 

  6. Shiraga H, Min W, VanDusen WJ, Clayman MD, Miner D, Terrell CH, Sherbotie JR, Foreman JW, Przysiecki C, Neilson EG, Hoyer JR (1992) Inhibition of calcium oxalate crystal growth in vitro by uropontin: another member of the aspartic acid-rich protein superfamily. Proc Natl Acad Sci USA 89:426–430

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Xie Y, Sakatsume M, Nishi S, Narita I, Arakawa M, Gejyo F (2001) Expression, roles, receptors, and regulation of osteopontin in the kidney. Kidney Int 60:1645–1657

    Article  CAS  PubMed  Google Scholar 

  8. Asplin JR, Arsenault D, Parks JH, Coe FL, Hoyer JR (1998) Contribution of human uropontin to inhibition of calcium oxalate crystallization. Kidney Int 53:194–199

    Article  CAS  PubMed  Google Scholar 

  9. Min W, Shiraga H, Chalko C, Goldfarb S, Krishna GG, Hoyer JR (1998) Quantitative studies of human urinary excretion of uropontin. Kidney Int 53:189–193

    Article  CAS  PubMed  Google Scholar 

  10. Wesson JA, Worcester EM, Wiessner JH, Mandel NS, Kleinman JG (1998) Control of calcium oxalate crystal structure and cell adherence by urinary macromolecules. Kidney Int 53:952–957

    Article  CAS  PubMed  Google Scholar 

  11. Lieske JC, Leonard R, Toback FG (1995) Adhesion of calcium oxalate monohydrate crystals to renal epithelial cells is inhibited by specific anions. Am J Physiol Renal Physiol 268:F604–F612

    CAS  Google Scholar 

  12. Yamate T, Kohri K, Umekawa T, Amasaki N, Amasaki N, Isikawa Y, Iguchi M, Kurita T (1996) The effect of osteopontin on the adhesion of calcium oxalate crystals to Madin-Darby canine kidney cells. Eur Urol 30:388–393

    CAS  PubMed  Google Scholar 

  13. Yamate T, Kohri K, Konya E, Ishikawa Y, Iguchi M, Kurita T (1999) Interaction between osteopontin on Madin Darby Canine Kidney cell membrane and calcium oxalate crystal. Urol Int 62:81–86

    Article  CAS  PubMed  Google Scholar 

  14. Wesson JA, Johnson RJ, Mazzali M, Beshensky AM, Stietz S, Giachelli C, Liaw L, Alpers CE, Couser WG, Kleinman JG, Hughes J (2003) Osteopontin is a critical inhibitor of calcium oxalate crystal formation and retention in renal tubules. J Am Soc Nephrol 14:139–147

    Article  CAS  PubMed  Google Scholar 

  15. Kohri K, Nomura S, Kitamura Y, Nagata T, Yoshioka K, Iguchi M, Yamate T, Umekawa T, Suzuki Y, Sinohara H, Kurita T (1993) Structure and expression of the mRNA encoding urinary stone protein (osteopontin). J Biol Chem 268:15180–15184

    CAS  PubMed  Google Scholar 

  16. Bautista DS, Denstedt J, Chambers AF (1996) Low-molecular-weight variants of osteopontin generated by serine proteinases in urine of patients with kidney stones. J Cell Biochem 61:402–409

    Article  CAS  PubMed  Google Scholar 

  17. Hoyer JR, Pietrzyk RA, Liu H, Whitson PA (1999) Effects of microgravity on urinary osteopontin. Am Soc Nephrol 10:S389–393

    CAS  Google Scholar 

  18. Nishio S, Hatanaka M, Takeda H, Iseda T, Iwata H, Yokoyama M (1999) Analysis of urinary concentrations of calcium phosphate crystal-associated proteins: alpha2-HS-glycoprotein, prothrombin F1, and osteopontin. J Am Soc Nephrol 10:S394–S396

    CAS  PubMed  Google Scholar 

  19. Kon S, Maeda M, Segawa T, Hagiwara Y, Horikoshi Y, Chikuma S, Tanaka K, Rashid MM, Inobe M, Chambers AF, Uede T (2000) Antibodies to different peptides in osteopontin reveal complexities in the various secreted forms. J Cell Biochem 77:487–498

    Article  CAS  PubMed  Google Scholar 

  20. Yasui T, Fujita K, Hayashi Y, Ueda K, Kon S, Maeda M, Uede T, Kohri K (1999) Quantification of osteopontin in the urine of healthy and stone-forming men. Urol Res 27:225–230

    Article  CAS  PubMed  Google Scholar 

  21. Dean CJ, Macardle PJ, Ryall RL (2000) The effect of the presence of calcium oxalate crystals on the measurement of prothrombin fragment 1 in urine. In: Rodgers AL, Hibbert BE, Hess B, Khan SR, Preminger GM (eds) Urolithiasis 200. University of Cape Town, Cape Town, pp 150–152

    Google Scholar 

  22. Ryall RL, Chauvet MC, Grover PK (2005) Intracrystalline proteins and urolithiasis: a comparison of the protein content and ultrastructure of urinary calcium oxalate monohydrate and dihydrate crystals. BJU Int 96:654–663

    Article  CAS  PubMed  Google Scholar 

  23. Qiu SR, Wierzbicki A, Orme CA, Cody AM, Hoyer JR, Nancollas GH, Zepeda S, De Yoreo JJ (2004) Molecular modulation of calcium oxalate crystallization by osteopontin and citrate. Proc Nat Acad Sci USA 101:1811–1815

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Brown P, Ackermann D, Finlayson B (1989) Calcium oxalate dihydrate (weddellite) precipitation. J Cryst Growth 98:285–292

    Article  CAS  Google Scholar 

  25. Ryall RL, Hibberd CM, Marshall VR (1985) A method for studying inhibitory activity in whole urine. Urol Res 13:285–289

    Article  CAS  PubMed  Google Scholar 

  26. Lieske JC, Hammers MS, Hoyer JR, Toback FG (1997) Renal cell osteopontin production is stimulated by calcium oxalate monohydrate crystals. Kidney Int 51:679–686

    Article  CAS  PubMed  Google Scholar 

  27. Huang H-S, Ming-Chieh MA, Chen C-F, Chen J (2003) Lipid peroxidation and its correlations with urinary levels of oxalate, citric acid, and osteopontin in patients with renal calcium oxalate stones. Urology 62:1123–1128

    Article  PubMed  Google Scholar 

  28. Nishio S, Hatanaka M, Takeda H, Aoki K, Iseda H, Yokoyama M (2001) Calcium phosphate crystal-associated proteins: alpha-2-HS-glycoprotein, prothrombin fragment 1 and osteopontin. Int J Urol 8:S58–S62

    Article  CAS  PubMed  Google Scholar 

  29. Ryall RL, Grover PK, Thurgood LA, Chauvet MC, Fleming DE, van Bronswijk W (2007) The importance of a clean face: the effect of different washing procedures on the association of Tamm–Horsfall glycoprotein and other urinary proteins with calcium oxalate crystals. Urol Res 35:1–14

    Article  CAS  PubMed  Google Scholar 

  30. Meyer JL, Smith LH (1975) Growth of calcium oxalate crystals. II. Inhibition by natural urinary crystal growth inhibitors. Invest Urol 13:36–39

    CAS  PubMed  Google Scholar 

  31. Kazanecki CC, Kowalski AJ, Ding T, Rittling SR, Denhardt DT (2007) Characterization of anti-osteopontin monoclonal antibodies: binding sensitivity to post-translational modifications. J Cell Biochem 102:925–935

    Article  CAS  PubMed  Google Scholar 

  32. Kleinman JG, Wesson JA, Hughes J (2004) Osteopontin and calcium stone formation. Nephron Physiol 98:43–47

    Article  Google Scholar 

  33. Krakauer T (1998) Variability in the sensitivity of nine enzyme-linked immunosorbant assays (ELISAs) in the measurement of human interleukin 6. J Immunol Methods 219:161–167

    Article  CAS  PubMed  Google Scholar 

  34. Vordermark D, Said HM, Katzer A, Kuhnt T, Hänsgen G, Dunst J, Flentje M, Bache M (2006) Plasma osteopontin levels in patients with head and neck cancer and cervix cancer are critically dependent on the choice of ELISA system. BMC Cancer 6:207

    Article  PubMed  PubMed Central  Google Scholar 

  35. Hoyer JR, Asplin JR, Otvos L (2001) Phosphorylated osteopontin peptides suppress crystallization by inhibiting the growth of calcium oxalate crystals. Kidney Int 60:77–82

    Article  CAS  PubMed  Google Scholar 

  36. Addadi L, Weiner S (1985) Interactions between acidic proteins and crystals: stereochemical requirements in biomineralization. Proc Natl Acad Sci USA 82:4110–4114

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Yamaguchi H, Igarashi M, Hirata A, Tsuchiya H, Sugiyama K, Morita Y, Jimbu Y, Ohnuma H, Daimon M, Tomonaga M, Kato T (2004) Progression of diabetic nephropathy enhances the plasma osteopontin level in type 2 diabetic patients. Endocr J 51:499–504

    Article  CAS  PubMed  Google Scholar 

  38. Gang X, Ueki K, Kon S, Maeda M, Naruse T, Nojima Y (2001) Reduced urinary excretion of intact osteopontin in patients with IgA nephropathy. Am J Kidney Dis 37:374–379

    Article  CAS  PubMed  Google Scholar 

  39. Ye B, Skates S, Mok SC, Horick NK, Rosenberg HF, Vitonis A, Edwards D, Sluss P, Han WK, Berkowitz RS, Cramer DW (2006) Proteomic-based discovery and characterization of glycosylated eosinophil-derived neurotoxin and COOH-terminal osteopontin fragments for ovarian cancer in urine. Clin Cancer Res 12:432–441

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Funding for this study was provided by National Institute of Diabetes and Digestive and Kidney Diseases Grant 1R01-DK-064050-01A1.

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Authors and Affiliations

  1. Department of Surgery, Flinders Medical Centre and Flinders University School of Medicine, Bedford Park, SA, 5042, Australia

    Lauren A. Thurgood, Phulwinder K. Grover & Rosemary Lyons Ryall

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  1. Lauren A. Thurgood
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  2. Phulwinder K. Grover
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  3. Rosemary Lyons Ryall
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Correspondence to Rosemary Lyons Ryall.

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Thurgood, L.A., Grover, P.K. & Ryall, R.L. High calcium concentration and calcium oxalate crystals cause significant inaccuracies in the measurement of urinary osteopontin by enzyme linked immunosorbent assay. Urol Res 36, 103–110 (2008). https://doi.org/10.1007/s00240-008-0139-3

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  • DOI: https://doi.org/10.1007/s00240-008-0139-3

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