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Proteomic Identification of Urinary Biomarkers of Diabetic Nephropathy

¹ Department of Endocrinology and Metabolism, Nizam’s Institute of Medical Sciences University, Hyderabad, India
² Department of Pediatrics, Oregon Health and Science University, Portland, Oregon
³ Department of Nephrology, Nizam’s Institute of Medical Sciences University, Hyderabad, India

Address correspondence and reprint requests to Srinivasa R. Nagalla, MD, Pediatrics (NRC5), Oregon Health and Science University, 3181 S.W. Sam Jackson Park Rd., Portland, OR 97239-3098. E-mail: nagallas{at}ohsu.edu

OBJECTIVE—Diabetic nephropathy is a serious complication of both type 1 and type 2 diabetes, and, unless arrested, leads to end-stage renal disease. Current diagnosis consists of urine assays of microalbuminuria, which have inadequate specificity and sensitivity.

RESEARCH DESIGN AND METHODS—We used proteomic analyses to identify novel biomarkers of nephropathy in urine from type 2 diabetic patients with demonstrated normo-, micro-, or macroalbuminuria. Samples were analyzed by fluorescence two-dimensional (2-D) differential in-gel electrophoresis (DIGE), and protein identification was performed by liquid chromatography-tandem mass spectrometry.

RESULTS—2-D DIGE analysis of the urinary proteome in diabetes with nephropathy identified 195 protein spots representing 62 unique proteins. These proteins belonged to several functional groups, i.e., cell development, cell organization, defense response, metabolism, and signal transduction. Comparisons between control and diabetic subjects with different stages of renal dysfunction revealed the differential expression of several proteins. Spot volume quantification identified 7 proteins that were progressively upregulated with increasing albuminuria and 4 proteins that exhibited progressive downregulation. The majority of these potential candidate biomarkers were glycoproteins.

CONCLUSIONS—These data demonstrate the ability of proteomic analyses to reveal potential biomarkers for diabetic nephropathy in urine, an important step forward in advancing accurate diagnosis and our understanding of disease mechanisms.

Abbreviations: A1AT, ₁-antitrypsin • AMBP, ₁-microglobulin/bikunin precursor • DIGE, differential in-gel electrophoresis • ESRD, end-stage renal disease • LC, liquid chromatography • MS, mass spectrometry • MS/MS, tandem mass spectroscopy • RBP, retinol-binding protein • VDBP, vitamin D–binding protein

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