Urinary Peptidomic Analysis Identifies Potential Biomarkers for Acute Rejection of Renal Transplantation
Human urine is a complex matrix of proteins, endogenous peptides, lipids, and metabolites. The level of any or all of these components can reflect the pathophysiological status of an individual especially of the kidney at the time of urine collection. The naturally occurring endogenous urinary peptides which are thought to be the product of several proteolytic and degradation processes may provide clinically useful biomarkers for different renal and systemic diseases.
Materials and Methods
To examine if specific differences in the urinary peptidome (<10 kDa) occur at the time of acute renal transplant rejection (AR), we undertook a study of urine samples collected from biopsy-proven AR (n = 10), stable graft function (n = 10), and healthy normal control (n = 10). The peptides (<10 kDa) were extracted and fractionated with high-performance liquid chromatography followed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometric (MS) analysis.
We identified 54 endogenous peptides, including multiple peptides for Tamm–Horsfall protein (UMOD). A panel of peptides are identified which discriminate renal transplant patients with AR from stable graft. We have shown that liquid chromatography followed by MALDI is a useful tool to identify potential biomarkers, which after verification with larger patient cohort can be used as a non-invasive monitoring tool for renal transplant rejection.
- Pisitkun T, Johnstone R, Knepper MA. Discovery of urinary biomarkers. Mol Cell Proteomics. 2006;5:1760–71. CrossRef
- Veenstra TD, Conrads TP, Hood BL, Avellino AM, Ellenbogen RG, Morrison RS. Biomarkers: mining the biofluid proteome. Mol Cell Proteomics. 2005;4:409–18. CrossRef
- Gulanikar AC, MacDonald AS, Sungurtekin U, Belitsky P. The incidence and impact of early rejection episodes on graft outcome in recipients of first cadaver kidney transplants. Transplantation. 1992;53:323–8. CrossRef
- Hariharan S, Johnson CP, Bresnahan BA, Taranto SE, McIntosh MJ, Stablein D. Improved graft survival after renal transplantation in the United States, 1988 to 1996. N Engl J Med. 2000;342:605–12. CrossRef
- Vidhun J, Masciandro J, Varich L, Salvatierra O Jr, Sarwal M. Safety and risk stratification of percutaneous biopsies of adult-sized renal allografts in infant and older pediatric recipients. Transplantation. 2003;76:552–7. CrossRef
- Zimmerli LU, Schiffer E, Zurbig P, et al. Urinary proteomic biomarkers in coronary artery disease. Mol Cell Proteomics. 2008;7:290–8.
- Wu DL, Zhang WH, Wang WJ, Jing SB, Xu YM. Proteomic Evaluation of Urine from Renal Cell Carcinoma Using SELDI-TOF-MS and Tree Analysis Pattern. Technol Cancer Res Treat. 2008;7:155–60.
- Woodson K, O’Reilly KJ, Hanson JC, Nelson D, Walk EL, Tangrea JA. The usefulness of the detection of GSTP1 methylation in urine as a biomarker in the diagnosis of prostate cancer. J Urol. 2008;179:508–11. discussion 11-2. CrossRef
- Webb KS, Lin GH. Urinary fibronectin: potential as a biomarker in prostatic cancer. Invest Urol. 1980;17:401–4.
- Thrailkill KM, Bunn RC, Moreau CS, et al. Matrix metalloproteinase-2 dysregulation in type 1 diabetes. Diabetes Care. 2007;30:2321–6. CrossRef
- Clarke W, Silverman BC, Zhang Z, Chan DW, Klein AS, Molmenti EP. Characterization of renal allograft rejection by urinary proteomic analysis. Ann Surg. 2003;237:660–664. discussion 4-5. CrossRef
- Vidal BC, Bonventre JV, I-HongHsu S. Towards the application of proteomics in renal disease diagnosis. Clin Sci(Lond). 2005;109:421–30. CrossRef
- Oetting WS, Rogers TB, Krick TP, Matas AJ, Ibrahim HN. Urinary beta2-microglobulin is associated with acute renal allograft rejection. Am J Kidney Dis. 2006;47:898–904. CrossRef
- Voshol H, Brendlen N, Muller D, et al. Evaluation of biomarker discovery approaches to detect protein biomarkers of acute renal allograft rejection. J Proteome Res. 2005;4:1192–9. CrossRef
- Parikh CR, Jani A, Mishra J, et al. Urine NGAL and IL-18 are predictive biomarkers for delayed graft function following kidney transplantation. Am J Transplant. 2006;6:1639–45. CrossRef
- Zhou H, Pisitkun T, Aponte A, et al. Exosomal Fetuin-A identified by proteomics: a novel urinary biomarker for detecting acute kidney injury. Kidney Int. 2006;70:1847–57. CrossRef
- Wittke S, Haubitz M, Walden M, et al. Detection of acute tubulointerstitial rejection by proteomic analysis of urinary samples in renal transplant recipients. Am J Transplant. 2005;5:2479–88. CrossRef
- Decramer S, Gonzalezdeperedo A, Breuil B, et al. Urine in clinical proteomics. Mol Cell Proteomics. 2008;7(10):1850–62. CrossRef
- Jurgens M, Appel A, Heine G, et al. Towards characterization of the human urinary peptidome. Comb Chem High Throughput Screen. 2005;8:757–65. CrossRef
- Fiedler GM, Baumann S, Leichtle A, et al. Standardized peptidome profiling of human urine by magnetic bead separation and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Clin Chem. 2007;53:421–8. CrossRef
- Schwartz GJ, Brion LP, Spitzer A. The use of plasma creatinine concentration for estimating glomerular filtration rate in infants, children, and adolescents. Pediatr Clin North Am. 1987;34:571–90.
- Sigdel TK, Lau K, Schilling J, Sarwal M. Optimizing protein recovery for urinary proteomics, a tool to monitor renal transplantation. Clin Transplant. 2008;22:617–23.
- Snyder SL, Sobocinski PZ. An improved 2, 4, 6-trinitrobenzenesulfonic acid method for the determination of amines. Analytical Biochemistry. 1975;64:284–8. CrossRef
- Tibshirani R, Hastie T, Narasimhan B, et al. Sample classification from protein mass spectrometry, by ‘peak probability contrasts’. Bioinformatics. 2004;20:3034–44. CrossRef
- Yasui Y, Pepe M, Thompson ML, et al. A data-analytic strategy for protein biomarker discovery: profiling of high-dimensional proteomic data for cancer detection. Biostatistics. 2003;4:449–63. CrossRef
- Habeeb AF. Determination of free amino groups in proteins by trinitrobenzenesulfonic acid. Anal Biochem. 1966;14:328–36. CrossRef
- Adachi J, Kumar C, Zhang Y, Olsen JV, Mann M. The human urinary proteome contains more than 1500 proteins, including a large proportion of membrane proteins. Genome Biol. 2006;7:R80. CrossRef
- Herrero M, Ibanez E, Cifuentes A. Capillary electrophoresis-electrospray-mass spectrometry in peptide analysis and peptidomics. Electrophoresis. 2008;29(10):2148–60. CrossRef
- Jacobs JM, Adkins JN, Qian WJ, et al. Utilizing human blood plasma for proteomic biomarker discovery. J Proteome Res. 2005;4:1073–85. CrossRef
- Kanie Y, Enomoto A, Goto S, Kanie O. Comparative RP-HPLC for rapid identification of glycopeptides and application in off-line LC-MALDI-MS analysis. Carbohydr Res. 2008;343:758–68. CrossRef
- Stodulkova E, Novak P, Deininger SO, et al. LC MALDI-TOF MS/MS and LC ESI FTMS analyses of HLA-B27 associated peptides isolated from peripheral blood cells. Immunol Lett. 2008;116:79–85.
- Sadiq ST, Agranoff D. Pooling serum samples may lead to loss of potential biomarkers in SELDI-ToF MS proteomic profiling. Proteome Sci. 2008;6:16. CrossRef
- Khan A, Packer NH. Simple urinary sample preparation for proteomic analysis. J Proteome Res. 2006;5:2824–38. CrossRef
- Zhou H, Yuen PS, Pisitkun T, et al. Collection, storage, preservation, and normalization of human urinary exosomes for biomarker discovery. Kidney Int. 2006;69:1471–6.
- Schaub S, Wilkins JA, Antonovici M, et al. Proteomic-based identification of cleaved urinary beta2-microglobulin as a potential marker for acute tubular injury in renal allografts. Am J Transplant. 2005;5:729–38. CrossRef
- Vio CP, Olavarria V, Gonzalez C, Nazal L, Cordova M, Balestrini C. Cellular and functional aspects of the renal kallikrein system in health and disease. Biol Res. 1998;31:305–22. CrossRef
- Villanueva J, Shaffer DR, Philip J, et al. Differential exoprotease activities confer tumor-specific serum peptidome patterns. J Clin Invest. 2006;116:271–84. CrossRef
- Urinary Peptidomic Analysis Identifies Potential Biomarkers for Acute Rejection of Renal Transplantation
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
Volume 5, Issue 2 , pp 103-113
- Cover Date
- Print ISSN
- Online ISSN
- Humana Press Inc
- Additional Links
- Renal transplantation
- LC MALDI
- Urinary Peptidomics
- Acute rejection
- Non-invasive biomarkers
- Biomarker discovery
- Author Affiliations
- 1. Division of Nephrology, Department of Pediatrics, Stanford University Medical School, Stanford University, Stanford, CA, 94305, USA
- 2. Divisions of Immunology, Department of Pediatrics, Stanford University Medical School, Stanford University, Stanford, CA, 94305, USA
- 3. Stanford University Medical School, G306, 300 Pasteur Drive, Stanford, CA, 94304, USA