Proteomics in Ménière disease

Abstract

Ménière's disease (MD) is a disorder of the inner ear characterized by an insidious onset and aspecific symptoms, such as dizziness, vertigo, tinnitus, and hearing loss, that may become very debilitating. The presence of endolymphatic hydrops is a common feature in MD patients, but the pathophysiology is still largely unknown. In this study, we have used a proteomics-driven approach to identify potential biomarkers of MD. To this end, plasma was obtained from whole blood of 16 individuals previously diagnosed as suffering from MD and compared to plasma from healthy donors. A depletion of the highly abundant proteins (i.e., albumin, IgG, transferrin, etc.) was performed in order to enhance the chance of detection of the less represented ones, therefore reducing the noise-background. Two-dimensional gel electrophoresis, followed by in-gel tryptic digestion of the selected spots and LC-MS/MS analysis, allowed us to identify a set of proteins whose expression appears to be differentially modulated in patients versus controls. In particular: complement factor H and B, fibrinogen alpha and gamma chains, beta-actin and pigment epithelium derived factor are over expressed; on the other hand, the levels of beta-2 glycoprotein-1, vitamin D binding protein and apolipoprotein-1 are significantly decreased in the plasma of MD-affected individuals. Even though preliminary and not necessarily linked directly to the molecular pathogenesis of the disease, our original findings suggest that a molecular signature, represented by the plasma protein profile previously described, might represent a potentially powerful, innovative and not invasive tool for early diagnosis and clinical management of MD patients. J. Cell. Physiol. 227: 308-312, 2012. © 2011 Wiley Periodicals, Inc.

A Proteomics‐Driven Assay Defines Specific Plasma Protein Signatures in Different Stages of Ménière's Disease

“…Figure 1 shows the results of the densitometric analysis. Interestingly, the trend of expression of the plasma proteins probed in our experimental setting fully confirmed the findings reported in our previous work [Chiarella and Saccomanno, 2012]. In detail, complement factor-B and C, fibrinogen a-chain, b-actin, pigment epithelium-derived factor and fibrinogen g-chain were found to be upregulated in MD-affected patients with respect to the healthy donors, while vitamin D-binding protein, apolipoprotein A-1 and b-2-glycoprotein were underexpressed in patients versus controls plasma.…”

“…Depletion of highly abundant protein from whole plasma was performed as previously described [Chiarella and Saccomanno, 2012]. Briefly, each sample was subjected to the Proteome-Lab TM IgY-12…”

“…Depletion of highly abundant protein from whole plasma was performed as previously described [Chiarella and Saccomanno, 2012]. Briefly, each sample was subjected to the Proteome-Lab TM IgY-12 Proteome Partitioning Kit (Beckman Coulter, Crea, CA) according to the manufacturer 0 s specifications in order to remove the 12 most abundant proteins (albumin, total IgGs, a1-antitrypsin, IgA, IgM, transferrin, haptoglobin, a1-acid glycoprotein, a2-macroglobulin, HDL-binding apolipoproteins A-1 and A-2, fibrinogen).…”

“…In the present study, we extended the analysis of our previous work [Chiarella and Saccomanno, 2012], aimed at evaluating the proteomic pattern of plasma proteins from patients suffering from MD. Moreover, we have attempted to correlate specific protein profiles with the stage of the disease.…”

Unraveling the Mechanistic Complexity of the Glomerulocystic Phenotype in Dicer Conditional KO Mice by 2D Gel Electrophoresis Coupled Mass Spectrometry

“…Second dimension separation was run on 10% SDS‐polyacrylamide gels, (2 W gel; 25°C) until the bromophenol blue dye front reached the end of the gels . Gels were stained with MS compatible silver staining procedure . Each sample analysis was performed in triplicate.…”