title: Proteomic identification of vanin-1 as a marker of kidney damage in a rat model of type 1 diabetic nephropathy.
abstract: At present, the urinary albumin excretion rate is the best noninvasive predictor for diabetic nephropathy (DN) but major limitations are associated with this marker. Here, we used in vivo perfusion technology to establish disease progression markers in an animal model of DN. Rats were perfused with a reactive ester derivative of biotin at various times after streptozotocin treatment. Following homogenization of kidney tissue and affinity purification of biotinylated proteins, a label-free mass spectrometry-based proteomic analysis of tryptic digests identified and relatively quantified 396 proteins. Of these proteins, 24 and 11 were found to be more than 10-fold up- or downregulated, respectively, compared with the same procedure in vehicle-treated rats. Changes in the expression of selected differentially regulated proteins were validated by immunofluorescence detection in kidney tissue from control and diabetic rats. Immunoblot analysis of pooled human urine found that concentrations of vanin-1, an ectoenzyme pantetheinase, distinguished diabetic patients with macroalbuminuria from those with normal albuminuria. Uromodulin was elevated in the urine pools of the diabetic patients, regardless of the degree of albuminuria, compared with healthy controls. Thus, in vivo biotinylation facilitates the detection of disease-specific changes in the abundance of potential biomarker proteins for disease monitoring and/or pharmacodelivery applications.
authors.affiliation: Department of Chemistry and Applied Biosciences, Institute of Pharmaceutical Sciences, ETH Zurich, Zurich, Switzerland.
authors.forename: Tim
authors.initials: T
authors.lastname: Fugmann
authors.forename: Beatrice
authors.initials: B
authors.lastname: Borgia
authors.forename: Csaba
authors.initials: C
authors.lastname: Révész
authors.forename: Mária
authors.initials: M
authors.lastname: Godó
authors.forename: Carol
authors.initials: C
authors.lastname: Forsblom
authors.forename: Peter
authors.initials: P
authors.lastname: Hamar
authors.forename: Harry
authors.initials: H
authors.lastname: Holthöfer
authors.forename: Dario
authors.initials: D
authors.lastname: Neri
authors.forename: Christoph
authors.initials: C
authors.lastname: Roesli
mesh_terms: D000419:Albuminuria / Q000175:diagnosis* / Q000201:enzymology / Q000209:etiology; D000581:Amidohydrolases / Q000378:metabolism* / Q000652:urine; D000818:Animals; D015415:Biomarkers / Q000378:metabolism / Q000652:urine; D019921:Biotinylation; D016022:Case-Control Studies; D002846:Chromatography, Affinity; D003921:Diabetes Mellitus, Experimental / Q000150:complications* / Q000201:enzymology; D003922:Diabetes Mellitus, Type 1 / Q000150:complications* / Q000201:enzymology; D003928:Diabetic Nephropathies / Q000175:diagnosis* / Q000201:enzymology / Q000209:etiology; D058851:GPI-Linked Proteins / Q000378:metabolism / Q000652:urine; D006801:Humans; D007668:Kidney / Q000201:enzymology*; D008297:Male; D013058:Mass Spectrometry; D010449:Peptide Mapping; D040901:Proteomics / Q000379:methods; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D058949:Uromodulin / Q000652:urine
chemical_list: D015415:Biomarkers; D058851:GPI-Linked Proteins; C548612:UMOD protein, human; D058949:Uromodulin; D000581:Amidohydrolases; C022314:pantetheinase