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Urine proteomic profiling of pediatric nephrotic syndrome

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Abstract

The prognosis of pediatric nephrotic syndrome (NS) correlates with the responsiveness to glucocorticoid therapy. Steroid-resistant NS (SRNS) patients progress to end-stage renal disease, while steroid-sensitive NS (SSNS) and steroid-dependent (SDNS) patients do not. We have performed proteomic profiling of urine samples from a cross section of pediatric and adolescent subjects with SSNS, SRNS, and orthostatic proteinuria (OP) to identify urinary biomarkers of steroid resistance. We performed surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS) on urine from 19 subjects with SSNS/SDNS in remission, 14 with SSNS/SDNS in relapse, 5 with SRNS in relapse, and 6 with OP. Genetic algorithm search of principal component space revealed a group of five peaks distinguishing SRNS subjects, with mass/charge (m/z) values of 3,917.07, 4,155.53, 6,329.68, 7,036.96, and 11,117.4. Our analyses identified the peak at m/z 11,117.4 with an accuracy of 95% for classifying SRNS. Multidimensional protein fractionation and mass spectrometric analysis of SRNS urine samples combined with immunodepletion identified the 11,117.4 protein as β2-microglobulin (B2M). Using an unbiased protein profiling approach, we have validated previously reported findings of B2M as a biomarker associated with SRNS. Prospective studies are warranted to establish additional biomarkers that would be predictive of SRNS.

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Abbreviations

B2M:

β2-Microglobulin

FSGS:

Focal segmental glomerulosclerosis

GA:

Genetic algorithm

HPRP:

High-performance reverse phase

MALDI:

Matrix-assisted laser desorption/ionization

MS:

Mass spectrometry

m/z :

Mass/charge

NS:

Nephrotic syndrome

OP:

Orthostatic proteinuria

PCA:

Principal component analysis

SD:

Steroid dependent

SDNS:

Steroid-dependent nephrotic syndrome

SELDI:

Surface-enhanced laser desorption/ionization

SR:

Steroid resistant

SRNS:

Steroid-resistant nephrotic syndrome

SS:

Steroid sensitive

SSNS:

Steroid-sensitive nephrotic syndrome

TOF:

Time of flight

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Acknowledgements

This work was supported in part by NIH Training Grant T32 DK 007726 (MK and AZT), and by NIH grant K23 RR 16080 (ADS). Parts of this work were presented (abstract TH-PO160) at the American Society of Nephrology Renal Week 2005, Philadelphia, PA.

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Authors and Affiliations

  1. Department of Pediatrics, Division of Nephrology, Children’s Hospital Boston, Boston, MA, USA

    Mona Khurana, Avram Z. Traum & Asher D. Schachter

  2. Beth Israel Deaconess Medical Center, Boston, MA, USA

    Manuel Aivado, Meghan P. Wells, Manuel Guerrero, Franck Grall & Towia A. Libermann

  3. Dana Farber/Harvard Cancer Center Proteomics Core, Boston, MA, USA

    Manuel Aivado, Meghan P. Wells, Manuel Guerrero, Franck Grall & Towia A. Libermann

  4. Children’s Hospital Informatics Program at Harvard-MIT Division of Health Sciences and Technology, Boston, MA, USA

    Asher D. Schachter

  5. Harvard Medical School, Boston, MA, USA

    Mona Khurana, Avram Z. Traum, Manuel Aivado, Franck Grall, Towia A. Libermann & Asher D. Schachter

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  1. Mona Khurana
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  2. Avram Z. Traum
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  8. Asher D. Schachter
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Correspondence to Asher D. Schachter.

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MK and AZT contributed equally to this work.

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Khurana, M., Traum, A.Z., Aivado, M. et al. Urine proteomic profiling of pediatric nephrotic syndrome. Pediatr Nephrol 21, 1257–1265 (2006). https://doi.org/10.1007/s00467-006-0165-8

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