Proteomic profile of cerebrospinal fluid in patients with multiple sclerosis using two dimensional gel electrophoresis

Abstract

a faculty of medicine, department of clinical Pathology, alexandria university, alexandria, egypt; b chru de montpellier and université de montpellier, irmB, laboratoire de Biochimie Protéomique clinique, montpellier, france; c faculty of medicine, department of neuropsychiatry, alexandria university, alexandria, egypt; d faculty of medicine, department of anesthesia and surgical care, alexandria university, alexandria, egypt ABSTRACT Background: Multiple sclerosis (MS) is a highly heterogeneous disease affecting the central nervous system (CNS). Cerebrospinal fluid (CSF) has been preferred over other body fluids when studying this disease since it is the closest sample related to MS pathological changes. The aim of this study was to investigate differences of CSF protein expression in MS patients by twodimensional polyacrylamide gel electrophoresis (2-DE) in order to identify candidate biomarkers with potential clinical utility. Methods: We analysed the CSF protein expression of seven remitting-relapsing MS patients and seven control subjects using 2-DE. As a complementary step, the CSF of patients was pooled and compared to control-pooled CSF using 2-DE in order to minimise the inter-individual variations. Statistically significant spots were identified by mass spectrometry. Results: In the analysis of the individual samples, a consistent up-regulation of four statistically significant spots was noticed in the CSF of the patients. In the analysis of the pooled samples, 4 spots were up-regulated in the control pool, while 12 spots were up-regulated in the patient pool. Using matrix-assisted laser desorption ionisation with two time of lights (MALDI-TOF/TOF), we did not yield any results for the identification of proteins but with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), we were able to reach the identity of four proteins which were alpha-1-antichymotrypsin, prostaglandin-H2 D-isomerase, desmoplakin and hornerin. Conclusion:The results of this study support the presence of changes in the CSF proteome of MS patients. There is a need to investigate the role of these candidate CSF spots in a larger number of MS patients.

British Journal of Biomedical Science in 2016. What have we learned?

“…Due to the variety and non-specific nature of presenting signs and symptoms, the diagnosis of multiple sclerosis (MS) is hampered by lack of specific markers. Hassan and colleagues [22] used a proteomic approach with two-dimensional electrophoresis and MALDI-TOF/TOF mass spectroscopy to probe cerebro-spinal fluid from MS patients and controls. Four proteins (alpha-1-antichymotrypsin, prostaglandin-H2 D-isomerase, desmoplankin and hornerin) where present in significantly higher levels in the MS patients, suggesting these may be used as potential CSF markers of this disease.…”

Utility of Two-Dimensional Difference Gel Electrophoresis in Diagnosis of Multiple Sclerosis

“…In the present study, we aimed to analyze limitations of 2D-DIGE in proteomic studies of the CSF in order to explore possible reasons why results of previous studies in MS patients have been so controversial [ 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ]. It seems obvious using 2D-DIGE for identification of proteins in the CSF of MS patients.…”

“…After separation proteins can be picked from 2DE gels and identified e.g., by mass spectrometry. In MS, proteins of the cerebrospinal fluid (CSF) may be of major interest in order to identify possible candidate proteins which are up- or downregulated in comparison to healthy subjects [ 5 ]. There are some studies investigating CSF-proteins of MS patients by 2DE or two-dimensional difference gel electrophoresis (2D-DIGE) [ 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ].…”

“…In MS, proteins of the cerebrospinal fluid (CSF) may be of major interest in order to identify possible candidate proteins which are up- or downregulated in comparison to healthy subjects [ 5 ]. There are some studies investigating CSF-proteins of MS patients by 2DE or two-dimensional difference gel electrophoresis (2D-DIGE) [ 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 ]. 2D-DIGE provides the advantage of performing analyses on a reduced number of gels with comparison of samples in the same gel run under the same conditions and with a consistent pool-standard throughout all gels [ 14 , 15 , 16 ].…”

Proteomics of Multiple Sclerosis: Inherent Issues in Defining the Pathoetiology and Identifying (Early) Biomarkers

“…This review thus investigated whether currently identified protein species can be considered as biomarkers for MS, using comprehensive literature mining and bioinformatics to anchor the analysis. During the last two decades, proteomic approaches were used to assess various samples from animal models of MS (e.g., experimental autoimmune encephalomyelitis, EAE and cuprizone, CPZ), as well as from MS patients [ 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 , 92 , 93 , 94 , 95 , 96 , 97 , 98 , 99 , 100 , 101 , 102 , 103 , 104 , 105 ]. However, there do not appear to be any critical systematic reviews of the protein species identified from these studies.…”