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BMC Nephrol. 2015 Nov 17;16:190. doi: 10.1186/s12882-015-0185-3.

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

Author information

1
Department of Medicine, Division of Nephrology and Hypertension, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA. hogan.marie@mayo.edu.
2
Department of Medicine, Division of Nephrology and Hypertension, Mayo Clinic, 200 First Street SW, Rochester, MN, 55905, USA. Lieske.John@mayo.edu.
3
Department of Internal Medicine - Nephrology, University of Michigan Health System, Ann Arbor, MI, USA. NEPTUNE-Study@umich.edu.
4
Cardiovascular Research, Mayo Clinic, Rochester, MN, USA. Nesbitt.Lisa2@mayo.edu.
5
Pediatric Nephrology, University of Michigan, Ann Arbor, MI, USA. larysa@med.umich.edu.
6
Nephrology and Hypertension Research, Mayo Clinic, Rochester, MN, USA. Heyer.Christina@mayo.edu.
7
Nephrology and Hypertension Research, Mayo Clinic, Rochester, MN, USA. Harris.Peter@mayo.edu.
8
Department of Internal Medicine, Division of Nephrology and Hypertension, University of Kansas Medical Center, Kansas City, KS, USA. cward6@kumc.edu.
9
Nephrology and Hypertension Research, Mayo Clinic, Rochester, MN, USA. Sundsbak.Jamie@mayo.edu.
10
Oncology, Novartis Pharma AG, Basel, Switzerland. luca_manganelli@virgilio.it.
11
Department of Internal Medicine - Nephrology, University of Michigan Health System, Ann Arbor, MI, USA. wenjunj@umich.edu.
12
Kidney Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA. jeffreyk@mail.nih.gov.
13
Division of Nephrology and Kidney Research Institute, University of Washington, Seattle, WA, USA. nelsonpj@uw.edu.
14
Division of Nephrology and Hypertension, Harbor-UCLA Medical Center, Torrance, CA, USA. sadler@labiomed.org.
15
Department of Medicine, University Health Network and University of Toronto, Toronto, ON, Canada. heather.reich@uhn.ca.
16
Renal-Electrolyte and Hypertension Division, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. lholzman@mail.med.upenn.edu.
17
Department of Internal Medicine - Nephrology, University of Michigan Health System, Ann Arbor, MI, USA. kretzler@umich.edu.
18
Department of Internal Medicine - Nephrology, University of Michigan Health System, Ann Arbor, MI, USA. markusbi@umich.edu.

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.

PMID:
26577187
PMCID:
PMC4650313
DOI:
10.1186/s12882-015-0185-3
[Indexed for MEDLINE]
Free PMC Article

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