Comparative proteomic analysis of renal tissue in IgA nephropathy with iTRAQ quantitative proteomics

Abstract

Abstract. Immunoglobulin (Ig) A nephropathy (IgAN) is the most common form of glomerulonephritis. In clinical practice, it is difficult to monitor the repeating relapse in patients suffering from IgAN, which usually occurs within 10 years of end-stage renal disease. In order to identify and quantify the total protein content in the renal tissue of patients with IgAN, isobaric tags for relative and absolute quantification (iTRAQ) technology was performed. iTRAQ coupled with multiple chromatographic fractionation and tandem mass spectrometry was used to analyze the total protein of normal renal tissue in IgAN and healthy patients. The individual proteins were identified by the Mascot search engine and any that were differentially expressed were monitored. A total of 574 different proteins were identified, and 287 proteins were up-or downregulated by >1 fold alteration in levels. The results showed that iTRAQ-based quantitative proteomic technology for the identification and relative quantitation of the renal tissue proteome is efficiently applicable. The differential expression of the proteome profiles for IgAN patients was determined. Further studies using large cohorts of patient samples with long-term clinical follow-up data should be conducted to evaluate the usefulness of the pathogenesis and novel biomarker candidates of IgAN, which may develop a novel technique for the diagnosis of IgAN.

Analysis of the differential urinary protein profile in IgA nephropathy patients of Uygur ethnicity

“…As early as 2006, Mi-Ra Park et al used 2-DE and MALDI-TOF MS to identify the urinary proteins showing differential expression in IgAN and normal controls and determined that 59 proteins were differentially expressed in IgAN [7]. To date, many publications have reported differentially expressed proteins in the urine, urinary exosomes and kidney tissue of IgAN patients using various proteomic methods, including MALDI-TOF [4, 7, 8], 2D-GEL [1], iTRAQ [9, 10], SILAC [11], and label-free quantification methods [12, 13]. A total of 166 candidate biomarkers have been discovered, and some of these have been validated through ELISA or Western blot analysis.…”

Matching Kidneys and Urines: Establishing Noninvasive Surrogates of Intrarenal Events in Primary Glomerulonephritis

“…44 Emerging technologic advances in the field of proteomics suggests that the description of protein and peptide profiles in biopsy tissue likely will be a valuable and complementary platform for understanding tissue molecular responses. 45,46 This also opens the window to permit characterization of mRNA transcriptional regulation, protein-protein interactions, and other elements of protein metabolism and post-translational regulation (reviewed by Sigdel and Sarwal 47 ). Multiplex methods are evolving rapidly to facilitate rapid and sensitive quantitation of proteins in tissue samples, such as advanced approaches to mass spectrometry-based multiple reaction monitoring.…”

Proteomics and glomerulonephritis: A complementary approach in renal pathology for the identification of chronic kidney disease related markers

“…Additionally, these proteomic approaches are now capable of providing quantitative information when combined with analytical protocols based on stable isotope labeling or labelfree approaches [14]. In the former, extracted proteins are labeled with stable isotopes, heavy isotopes for one condition and light isotopes for another one.…”