Determination of ganglioside composition and structure in human brain hemangioma by chip-based nanoelectrospray ionization tandem mass spectrometry

Abstract

We report here on a preliminary investigation of ganglioside composition and structure in human hemangioma, a benign tumor in the frontal cortex (HFC) in comparison to normal frontal cortex (NFC) tissue using for the first time advanced mass spectrometric methods based on fully automated chip-nanoelectrospray (nanoESI) high-capacity ion trap (HCT) and collision-induced dissociation (CID). The high ionization efficiency, sensitivity and reproducibility provided by the chip-nanoESI approach allowed for a reliable MS-based ganglioside comparative assay. Unlike NFC, ganglioside mixture extracted from HFC was found dominated by species of short glycan chains exhibiting lower overall sialic acid content. In HFC, only GT1 (d18:1/20:0), and GT3 (d18:1/25:1) polysialylated species were detected. Interestingly, none of these trisialylated forms was detected in NFC, suggesting that such components might selectively be associated with HFC. Unlike the case of previously investigated high malignancy gliosarcoma, in HFC one modified O-Ac-GD2 and one modified O-Ac-GM4 gangliosides were observed. This aspect suggests that these O-acetylated structures could be associated with cerebral tumors having reduced malignancy grade. Fragmentation analysis by CID in MS(2) mode using as precursors the ions corresponding to GT1 (d18:1/20:0) and GD1 (d18:1/20:0) provided data corroborating for the first time the presence of the common GT1a and GT1b isomers and the incidence of unusual GT1c and GT1d glycoforms in brain hemangioma tumor.

A monophasic extraction strategy for the simultaneous lipidome analysis of polar and nonpolar retina lipids

“…Reconstituted retina lipid extracts were centrifuged at 2,000 g to remove any residual particulates, then transferred to 2.0 ml glass vials and stored at Ϫ 80°C until further use. Using this method, a total of three replicate polar upper phase and three replicate nonpolar lower phase retina lipid extracts were obtained, Numerous lipidomic methods for the characterization of gangliosides obtained from polar lipid extracts have been developed based on LC-MS platforms ( 23,49 ) as well as direct infusion by nESI ( 50,51 ), including the use of the highresolution/accurate mass Orbitrap platform ( 52 ). However, all ganglioside analysis methods reported to date have involved either the initial extraction and specifi c enrichment of gangliosides at the exclusion of other lipid classes by using variants of the classical Svennerholm ganglioside extraction method ( 17 ) or via the fractionation of total lipid extracts by chromatographic separation.…”

An Introduction to Sphingolipid Metabolism and Analysis by New Technologies

“…The high resolution and mass accuracy of the mass analyzer also allows a narrower precursor ion window, resulting in a better selection of lipid of interest (Haynes et al 2009). Chip-based nanoelectrospray ionization tandem mass spectrometry has demonstrated the capability of observing structural and composition differences of gangliosides (such as trisialoganglioside subspecies with different fatty acid chain lengths) between normal and benign tumorous frontal cortex tissues (Schiopu et al 2009). It has also been used to identify complex and distinctive gangliosides (such as GD, GT, GM, LM, GQ subspecies) in human gliosarcoma (Vukelic et al 2007).…”

Assessment of the Molecular Expression and Structure of Gangliosides in Brain Metastasis of Lung Adenocarcinoma by an Advanced Approach Based on Fully Automated Chip-Nanoelectrospray Mass Spectrometry

“…However, for the complete structural analysis of GSL, more sophisticated and advanced techniques are required. Presently, mass spectrometry (MS), intensively exploited for lipid characterization [32,33], is considered also one of the most precise and sensitive analytical methods in the detection and structural identification of different ganglioside species [34][35][36][37][38][39][40][41].…”

“…Thus, NanoMate robot coupled to either quadrupole time-of-flight (QTOF) or high capacity ion trap (HCT) mass spectrometer was introduced in GSL research for screening, sequencing, and identification of complex ganglioside mixtures extracted from normal [36][37][38] and pathologic human brain with a particular focus on primary brain tumors [35,40]. To continue this systematic mapping of gangliosides in brain tumors, in the present study, for the first time, chip-based nanoESI MS was employed for the investigation of ganglioside expression and structure in secondary brain tumors, i.e., brain metastasis of lung adenocarcinoma.…”