Identification and Characterization of New Proteins in Podocyte Dysfunction of Membranous Nephropathy by Proteomic Analysis of Renal Biopsy

Interstitial fluid, obtained by gentle centrifugation of the renal biopsy specimen, is highly enriched in elements directly secreted by the kidney tissue and is suitable for proteomic analysis. Here we describe the first clinical application of renal interstitial fluid analysis in a subset of samples obtained from patients affected by idiopathic membranous nephropathy. We included in the study fifty-one patients with different pathologic diagnoses. We identified the proteomic pattern of idiopathic membranous nephropathy with mass spectrometry analysis by comparing these samples with two controls: normal kidney and IgA nephropathy. Proteomic results were validated by immunofluorescence analysis of renal tissues and Western blot of serum, urines and podocyte cell cultures. We observed an increased expression of PDZ and LIM domain protein 5 (PDLI5) and LIM domain binding protein 3 (LDB3) providing first evidence of the differential expression of these LIM domain-related proteins in kidney and urines of patients with idiopathic membranous nephropathy. Interstitial fluid can be considered a valuable biological fluid in the discovery phase of biomarkers. In order to validate its clinical use, it is pivotal to assess the availability of the biomarkers in ‘usual’ samples: blood and/or urine. PDLI5 and LDB3 share a common LIM domain suggesting a possible role in the cytoskeleton organization and they appear upregulated in glomeruli of patients affected by idiopathic membranous nephropathy. Furthermore the two proteins become highly abundant in the urine of patients affected by idiopathic membranous nephropathy. In conclusion, our approach may be considered a novel method for identifying candidate biomarkers for patients suffering from membranous nephropathy and other glomerulonephrites