Contributions
Abstract: P542
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Biomarkers
Introduction: Cerebrospinal fluid (CSF) analysis is a powerful tool to identify specific biomarkers of multiple sclerosis (MS) diagnosis and prognosis. In addition to the study of inflammatory mediators involved in MS, a specific proteomic characterization of the presence and levels of proteins related to intrathecal inflammation and MS pathogenesis has been not fully examined.
Aims: In order to increase the diagnostic and prognostic role of CSF analysis in MS, we developed a new proteomic approach that allows deep examination of complex mixtures of proteins like CSF by identifying pathways probably involved in inflammation and tissue damage.
Methods: This study was performed on the CSF obtained by two MS populations stratified at time of diagnosis according to high (MSHigh) or low (MSLow) level of cortical lesion load detected by double inversion recovery 3T-MRI sequence and of CSF inflammation. CSFs from 3 MSHigh vs 3 MSLow patients were prepared by TRIDENT methodology by applying three-denaturation protocols, gradient SDS-PAGE separation and LC-MS/MS technologies. Bioinformatics analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID) and search tools for the retrieval of interacting genes (STRING) software, were carried out analysis of total number of proteins identified and related pathways.
Results: A total number of 227 proteins have been identified as expressed onto CSFs of the two examined groups of patients. Eleven main pathways have been identified as key mechanisms discriminating the MSHigh and MSlow patients.Among them, complement and coagulation cascade (29% of the total identified 227 proteins) was identified as the most modified pathway across the two groups of patients. ELISA methodology was then used for validation analyses, indicating that sCD163 (p< 0.0001), free hemoglobin (p< 0.05), haptoglobin (p< 0.0001) and fibrinogen (p< 0.01) were significantly higher in the CSF of MSHigh compared to MSLow patients. The protein expression of these molecules positively correlated with both number and volume of cortical lesions. On the contrary, the CSF levels of sCD14 were found significantly (p< 0.05) higher in MSLow compared to MSHigh and inversely correlated with cortical lesion load.
Conclusions: New proteomic CSF analysis approaches, including TRIDENT analysis, in combination with clinical and imaging assessment, may strongly contribute to shed light on molecular mechanisms of MS pathogenesis and severity/progression.
Disclosure: Roberta Magliozzi: nothing to disclose
Abstract: P542
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Biomarkers
Introduction: Cerebrospinal fluid (CSF) analysis is a powerful tool to identify specific biomarkers of multiple sclerosis (MS) diagnosis and prognosis. In addition to the study of inflammatory mediators involved in MS, a specific proteomic characterization of the presence and levels of proteins related to intrathecal inflammation and MS pathogenesis has been not fully examined.
Aims: In order to increase the diagnostic and prognostic role of CSF analysis in MS, we developed a new proteomic approach that allows deep examination of complex mixtures of proteins like CSF by identifying pathways probably involved in inflammation and tissue damage.
Methods: This study was performed on the CSF obtained by two MS populations stratified at time of diagnosis according to high (MSHigh) or low (MSLow) level of cortical lesion load detected by double inversion recovery 3T-MRI sequence and of CSF inflammation. CSFs from 3 MSHigh vs 3 MSLow patients were prepared by TRIDENT methodology by applying three-denaturation protocols, gradient SDS-PAGE separation and LC-MS/MS technologies. Bioinformatics analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID) and search tools for the retrieval of interacting genes (STRING) software, were carried out analysis of total number of proteins identified and related pathways.
Results: A total number of 227 proteins have been identified as expressed onto CSFs of the two examined groups of patients. Eleven main pathways have been identified as key mechanisms discriminating the MSHigh and MSlow patients.Among them, complement and coagulation cascade (29% of the total identified 227 proteins) was identified as the most modified pathway across the two groups of patients. ELISA methodology was then used for validation analyses, indicating that sCD163 (p< 0.0001), free hemoglobin (p< 0.05), haptoglobin (p< 0.0001) and fibrinogen (p< 0.01) were significantly higher in the CSF of MSHigh compared to MSLow patients. The protein expression of these molecules positively correlated with both number and volume of cortical lesions. On the contrary, the CSF levels of sCD14 were found significantly (p< 0.05) higher in MSLow compared to MSHigh and inversely correlated with cortical lesion load.
Conclusions: New proteomic CSF analysis approaches, including TRIDENT analysis, in combination with clinical and imaging assessment, may strongly contribute to shed light on molecular mechanisms of MS pathogenesis and severity/progression.
Disclosure: Roberta Magliozzi: nothing to disclose