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BMC Nephrology
Published in: BMC Nephrology 1/2015

Open Access 01-12-2015 | Research article

Strategy and rationale for urine collection protocols employed in the NEPTUNE study

Authors: Marie C. Hogan, John C. Lieske, Chrysta C. Lienczewski, Lisa L. Nesbitt, Larysa T. Wickman, Christina M. Heyer, Peter C. Harris, Christopher J. Ward, Jamie L. Sundsbak, Luca Manganelli, Wenjun Ju, Jeffrey B. Kopp, Peter J. Nelson, Sharon G. Adler, Heather N. Reich, Lawrence B. Holzmann, Matthias Kretzler, Markus Bitzer

Published in: BMC Nephrology | Issue 1/2015

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Abstract

Background

Glomerular diseases are potentially fatal, requiring aggressive interventions and close monitoring. Urine is a readily-accessible body fluid enriched in molecular signatures from the kidney and therefore particularly suited for routine clinical analysis as well as development of non-invasive biomarkers for glomerular diseases.

Methods

The Nephrotic Syndrome Study Network (NEPTUNE; ClinicalTrials.gov Identifier NCT01209000) is a North American multicenter collaborative consortium established to develop a translational research infrastructure for nephrotic syndrome. This includes standardized urine collections across all participating centers for the purpose of discovering non-invasive biomarkers for patients with nephrotic syndrome due to minimal change disease, focal segmental glomerulosclerosis, and membranous nephropathy. Here we describe the organization and methods of urine procurement and banking procedures in NEPTUNE.

Results

We discuss the rationale for urine collection and storage conditions, and demonstrate the performance of three experimental analytes (neutrophil gelatinase-associated lipocalin [NGAL], retinol binding globulin, and alpha-1 microglobulin) under these conditions with and without urine preservatives (thymol, toluene, and boric acid). We also demonstrate the quality of RNA and protein collected from the urine cellular pellet and exosomes.

Conclusions

The urine collection protocol in NEPTUNE allows robust detection of a wide range of proteins and RNAs from urine supernatant and pellets collected longitudinally from each patient over 5 years. Combined with the detailed clinical and histopathologic data, this provides a unique resource for exploration and validation of new or accepted markers of glomerular diseases.

Trial registration

ClinicalTrials.gov Identifier NCT01209000
Appendix
Available only for authorised users
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Metadata
Title
Strategy and rationale for urine collection protocols employed in the NEPTUNE study
Authors
Marie C. Hogan
John C. Lieske
Chrysta C. Lienczewski
Lisa L. Nesbitt
Larysa T. Wickman
Christina M. Heyer
Peter C. Harris
Christopher J. Ward
Jamie L. Sundsbak
Luca Manganelli
Wenjun Ju
Jeffrey B. Kopp
Peter J. Nelson
Sharon G. Adler
Heather N. Reich
Lawrence B. Holzmann
Matthias Kretzler
Markus Bitzer
Publication date
01-12-2015
Publisher
BioMed Central
Published in
BMC Nephrology / Issue 1/2015
Electronic ISSN: 1471-2369
DOI
https://doi.org/10.1186/s12882-015-0185-3

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