Naturally Occurring Human Urinary Peptides for Use in Diagnosis of Chronic Kidney Disease

Stefan Herget–Rosenthal ,Philippe Schmitt‐Kopplin ,Joshua J. Coon,Ziad A. Massy ,Petra Zürbig,Mario Luppi,Christian Neusüß ,Eric Schiffer,Eva M. Weissinger,Wilfried Gwinner,Markus Kellmann,Holger Jahn,Volker Kliem,Bruce A. Julian,Harald Rupprecht,Andrzej S. Królewski ,Harald Mischak,H Bauer ,Igor Golovko,Arnold Ganser,Joachim Jankowski,Walter Kölch ,Lise Tarnow,Georg Behrens,Anna F. Dominiczak,Christian Delles,Frank Eitner,George Jerums,Raymond Vanholder,Danilo Fliser,Stéphane Decramer,Karlheinz Peter,Dan Theodorescu,Marion Haubitz,Visith Thongboonkerd,J. H. H. Ehrich ,David W. Good ,Jan Novák ,Àngel Argilés,Moritz Frommberger,Joost P. Schanstra,Michael Melter,Mohammed Dakna,Kasper Rossing,Mark Girolami ,Jens‐Uwe Stolzenburg

doi:10.1074/mcp.m110.001917

Abstract

Because of its availability, ease of collection, and correlation with physiology and pathology, urine is an attractive source for clinical proteomics/peptidomics. However, the lack of comparable data sets from large cohorts has greatly hindered the development of clinical proteomics. Here, we report the establishment of a reproducible, high resolution method for peptidome analysis of naturally occurring human urinary peptides and proteins, ranging from 800 to 17,000 Da, using samples from 3,600 individuals analyzed by capillary electrophoresis coupled to MS. All processed data were deposited in an Structured Query Language (SQL) database. This database currently contains 5,010 relevant unique urinary peptides that serve as a pool of potential classifiers for diagnosis and monitoring of various diseases. As an example, by using this source of information, we were able to define urinary peptide biomarkers for chronic kidney diseases, allowing diagnosis of these diseases with high accuracy. Application of the chronic kidney disease-specific biomarker set to an independent test cohort in the subsequent replication phase resulted in 85.5% sensitivity and 100% specificity. These results indicate the potential usefulness of capillary electrophoresis coupled to MS for clinical applications in the analysis of naturally occurring urinary peptides.

Highlights

Chronic kidney disease (CKD)1 is often characterized by a slow, progressive loss of renal function with a loss of glomerular filtration over a period of months or years that may eventually lead to end stage renal disease (ESRD)
We report the application of CE-mass spectrometry (MS) for the analysis of the human urinary peptidome and demonstrate its utility for the definition of biomarkers of CKD in general
These biomarkers apparently reflect primary pathogenetic changes as well as the response to certain disease processes. Their usefulness extends beyond the applicability to diseases of the urogenital tract, and the approach may be applicable to diseases that result in systemic manifestations

Summary

Introduction

Application of the chronic kidney disease-specific biomarker set to an independent test cohort in the subsequent replication phase resulted in 85.5% sensitivity and 100% specificity These results indicate the potential usefulness of capillary electrophoresis coupled to MS for clinical applications in the analysis of naturally occurring urinary peptides. Renal damage is generally detected by proteinuria/albuminuria on urinalysis or quantitative measurement, changes in serum creatinine concentration for estimation of glomerular filtration rate, or both. These methods have major limitations as they are nonspecific and frequently are late manifestations of renal damage. Urine as a body fluid for clinical analysis is relatively stable, probably due to the fact that it is “stored” for hours in the bladder; proteolytic

Objectives

Results

Conclusion