Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease

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Abstract:

Background:

Sensitive and specific noninvasive biomarkers for tubulointerstitial injury are lacking, and proteomic techniques provide a powerful tool for biomarker discovery. Objective:

The aim of this study was to identify novel urinary biomarkers of early tubulointerstitial injury in canine progressive renal disease using both 2-dimensional differential in-gel electrophoresis (2-D DIGE), which identifies individual proteins, and surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF), which generates protein peak profiles. Methods:

Urine was collected from 6 male dogs with X-linked hereditary nephropathy (XLHN) at 2 time points (TP): 1) the onset of overt proteinuria (urine protein:creatinine ratio>2) and 2) the onset of azotemia (creatinine ≥1.2 mg/dL); corresponding renal biopsies were analyzed from 3 of the dogs. Urine samples from the 6 dogs were subjected to analysis by 2-D DIGE and SELDI-TOF. Urinary retinol-binding protein (RBP) was evaluated in 25 male dogs with XLHN and normal control dogs by Western blot analysis. Results:

Clinical data and histologic evaluation revealed reduced renal function and increased tubulointerstitial fibrosis at TP 2. A number of urine proteins and protein peaks were differentially present at the 2 time points, with several known biomarkers of renal disease identified in addition to several promising new biomarkers. RBP was first detected in urine approximately 2 months before onset of azotemia (TP 2), but after onset of overt proteinuria, and amounts increased with progression of disease. Conclusions:

Proteomic techniques were successfully used to identify urinary biomarkers of renal disease in dogs with XLHN. Urinary RBP is a promising biomarker for early detection of tubulointerstitial damage and progression to end-stage renal disease.

Keywords: 2-dimensional differential in-gel electrophoresis; Renal biomarker; XLHN; retinol-binding protein; surface-enhanced laser desorption ionization

Document Type: Research Article

DOI: http://dx.doi.org/10.1111/j.1939-165X.2011.00307.x

Affiliations: 1: Veterinary Pathobiology 2: Small Animal Clinical Sciences 3: Biochemistry and Biophysics, Texas A&M University, College Station, TX, USA 4: Department of Population Health and Pathobiology, North Carolina State University, Raleigh, NC, USA

Publication date: June 1, 2011

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