Abstract
In this preliminary investigation, a low-grade astrocytoma (AcT) is investigated by high-resolution (HR) mass spectrometry (MS) aiming at characterization of gangliosides with potential biomarker value. The research was conducted towards a comparative mapping of ganglioside expression in AcT, its surrounding tissue (ST) and a normal control brain tissue (NT). HR MS was conducted in the negative ion mode nanoelectrospray ionization (nanoESI). Fragmentation analysis was carried out by collision-induced dissociation (CID) MS2–MS4. Due to the high resolving power and mass accuracy, by comparative mapping of the ganglioside extracts from AcT, ST and NT, under identical conditions, 37 different species in AcT, 40 in ST and 56 in NT were identified. AcT and ST were found to contain 18 identical ganglioside components. Among all three specimens, ST extract presented the highest levels of sialylation, fucosylation and acetylation, a feature which might be correlated to the tumor expansion in the adjacent brain area. MS mapping indicated also that AcT, ST and NT share one doubly deprotonated molecule at m/z 1063.31, attributable to GT1(d18:1/18:0) or GT1(d18:0/18:1). CID MS2–MS4 on these particular ions detected in AcT and ST provided data supporting GT1c isomer in the investigated astrocytoma tissue. Our results show that HR MS has a remarkable potential in brain cancer research for the determination of tumor-associated markers and for their structural determination.
Similar content being viewed by others
Abbreviations
- AcT:
-
Astrocytoma
- CE:
-
Capillary electrophoresis
- Cer:
-
Ceramide
- CID:
-
Collision-induced dissociation
- ESI:
-
Electrospray ionization
- Fuc:
-
Fucose
- Gal:
-
Galactose
- GalNAc:
-
N-acetylgalactosamine
- Glc:
-
Glucose
- GlcNAc:
-
N-acetylglucosamine
- HCT:
-
High-capacity ion trap
- HPLC:
-
High-performance liquid chromatography
- HPTLC:
-
High-performance thin layer chromatography
- HR:
-
High resolution
- LC:
-
Liquid chromatography
- LCB:
-
Long chain base
- MALDI:
-
Matrix-assisted laser desorption/ionization
- MS:
-
Mass spectrometry
- MS/MS:
-
Tandem mass spectrometry
- MSn :
-
Multistage mass spectrometry
- NanoESI:
-
Nanoelectrospray ionization
- Neu5Ac:
-
N-acetyl neuraminic acid
- NT:
-
Normal brain tissue
- QTOF:
-
Quadrupole time-of-flight
- ST:
-
Surrounding tissue
- TIC:
-
Total ion chromatogram
- TLC:
-
Thin layer chromatography
- WHO:
-
World Health Organization
References
Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:97–109
IUPAC-IUB Joint Commission on Biochemical Nomenclature (1998) Nomenclature of glycolipids. Eur J Biochem 257:293–298
Greenberg MS (1997) Astrocytoma In: handbook of neurosurgery Vol 1 4th ed Lakeland, FL: Greenberg; 244–256
Louis David N, Hiroko O, Wiestler D, Otmar CK, Webster BC, Peter JA, Scheithauer Bernd W, Paul K (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 114:97–109
Wessels PH, Weber WE, Raven G, Ramaekers CS, Hopman HN, Twijnstra A (2003) Supratentorial grade II astrocytoma: biological features and clinical course. Lancet Neurol 2:395–403
Rodriguez FJ, Lim KS, Bowers D, Eberhart CG (2013) Pathological and molecular advances in pediatric low-grade astrocytoma. Annu Rev Pathol 8:361–379
Zhang H, Wu G, Tu H, Huang F (2007) Discovery of serum biomarkers in astrocytoma by SELDI-TOF MS and protein chip technology. J Neurooncol 84(3):315–323
Birkléa S, Zengb G, Gaob L, Yub RK, Aubryc J (2003) Role of tumor-associated gangliosides in cancer progression. Biochimie 85:455–459
Fernandez LE, Gabri MR, Guthmann MD, Gomez RE, Gold S, Fainboim L, Gomez DE, Alonso DF (2010) NGcGM3 Ganglioside: a privileged target for cancer vaccines. Clin Dev Immunol 2010:814397
Köhler M, Machill S, Salzer R, Krafft C (2009) Characterization of lipid extracts from brain tissue and tumors using Raman spectroscopy and mass spectrometry. Anal Bioanal Chem 393:1513–1520
Zhou Y, Liu CH, Sun Y, Pu Y, Boydston-White S, Liu Y, Alfano RR (2012) Human brain cancer studied by resonance Raman spectroscopy. J Biomed Opt 17:116021
Birks SM, Danquah JO, King L, Vlasak R, Gorecki DC, Pilkington GJ (2011) Targeting the GD3 acetylation pathway selectively induces apoptosis in glioblastoma. Neuro Oncol 13:950–960
Ali K, Lu Y, Das U, Sharma RK, Wiebe S, Meguro K, Sadanand V, Fourney DR, Vitali A, Kelly M, May T, Gomez J, Pellerin E (2010) Biomolecular diagnosis of human glioblastoma multiforme using Synchrotron mid-infrared spectromicroscopy. Int J Mol Med 26:11–16
Hwang J, Lee S, Lee JT, Kwon TK, Kim DR, Kim H, Park HC, Suk K (2010) Gangliosides induce autophagic cell death in astrocytes. Br J Pharmacol 159:586–603
Abate LE, Mukherjee P, Seyfried TN (2006) Gene-linked shift in ganglioside distribution influences growth and vascularity in a mouse astrocytoma. J Neurochem 98:1973–1984
Seyfried TN, Mukherjee P (2010) Ganglioside GM3 is antiangiogenic in malignant brain cancer. J Oncol 2010:961243
Vukelić Ž, Kalanj-Bognar S, Froesch M, Bîndila L, Radić B, Allen M, Peter-Katalinić J, Zamfir AD (2007) Human gliosarcoma-associated ganglioside composition is complex and distinctive as evidenced by high-performance mass spectrometric determination and structural characterization. Glycobiology 17:504–515
Schiopu C, Flangea C, Capitan F, Serb A, Vukelić Ž, Kalanj-Bognar S, Sisu E, Przybylski M, Zamfir AD (2009) Determination of ganglioside composition and structure in human brain hemangioma by chip-based nanoelectrospray ionization tandem mass spectrometry. Anal Bioanal Chem 395:2465–2477
Schiopu C, Vukelić Ž, Capitan F, Kalanj-Bognar S, Sisu E, Zamfir AD (2012) Chip-nanoelectrospray quadrupole time-of-flight tandem mass spectrometry of meningioma gangliosides: a preliminary study. Electrophoresis 33:1778–1786
Viljetić B, Labak I, Majić S, Stambuk A, Heffer M (2012) Distribution of mono-, di- and trisialo gangliosides in the brain of Actinopterygian fishes. Biochim Biophys Acta 1820:1437–1443
Kasperzyk JL, El-Abbadi MM, Hauser EC, D’Azzo A, Platt FM, Seyfried TN (2004) N-butyldeoxygalactonojirimycin reduces neonatal brain ganglioside content in a mouse model of GM1 gangliosidosis. J Neurochem 89:645–653
Furukawa K, Aixinjueluo W, Kasama T, Ohkawa Y, Yoshihara M, Ohmi Y, Tajima O, Suzumura A, Kittaka D, Furukawa K (2008) Disruption of GM2/GD2 synthase gene resulted in overt expression of 9-O-acetyl GD3 irrespective of Tis21. J Neurochem 105:1057–1066
Wagener R, Kobbe B, Stoffel W (1996) Quantification of gangliosides by microbore high-performance liquid chromatography. J Lipid Res 37:1823–1829
Whitehead SN, Chan KH, Gangaraju S, Slinn J, Li J, Hou ST (2011) Imaging mass spectrometry detection of gangliosides species in the mouse brain following transient focal cerebral ischemia and long-term recovery. PLoS One 6:e20808
Almeida R, Mosoarca C, Chirita M, Udrescu V, Dinca N, Vukelić Ž, Allen M, Zamfir AD (2008) Coupling of fully automated chip-based electrospray ionization to high-capacity ion trap mass spectrometer for ganglioside analysis. Anal Biochem 378:43–52
Zamfir AD, Flangea C, Altmann F, Rizzi A (2011) Glycosylation analysis of proteins, proteoglycans and glycolipids using capillary electrophoresis and mass spectrometry. Adv Chromatogr 49:135–194
Ikeda K, Taguchi R (2010) Highly sensitive localization analysis of gangliosides and sulfatides including structural isomers in mouse cerebellum sections by combination of laser microdissection and hydrophilic interaction liquid chromatography/electrospray ionization mass spectrometry with theoretically expanded multiple reaction monitoring. Rapid Commun Mass Spectrom 24:2957–2965
Ikegami T, Tomomatsu K, Takubo H, Horie K, Tanaka N (2008) Separation efficiencies in hydrophilic interaction chromatography. J Chromatogr A 1184:474–503
Haselberg R, De Jong GJ, Somsen GW (2011) Capillary electrophoresis-mass spectrometry for the analysis of intact proteins 2007–2010. Electrophoresis 32:66–82
Zamfir A, Vukelić Ž, Peter-Katalinić J (2002) A capillary electrophoresis and off-line capillary electrophoresis/electrospray ionization-quadrupole time of flight-tandem mass spectrometry approach for ganglioside analysis. Electrophoresis 23:2894–2903
Serb A, Schiopu C, Flagea C, Vukelić Ž, Sisu E, Zagrean L, Zamfir AD (2009) High-throughput analysis of gangliosides in defined regions of fetal brain by fully automated chip-based nanoelectrospray ionization multi-stage mass spectrometry. Eur J Mass Spectrom 15:541–553
Mosoarca C, Ghiulai RM, Novaconi CR, Vukelić Ž, Chiriac A, Zamfir AD (2011) Application of chip-based nanoelectrospray ion trap mass spectrometry to compositional and structural analysis of gangliosides in human fetal cerebellum. Anal Lett 44:1036–1049
Zamfir AD, Serb A, Vukelić Ž, Flangea C, Schiopu C, Marinčić D, Kalanj-Bognar S, Capitan F, Sisu E (2011) 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. J Am Soc Mass Spectrom 22:2145–2159
Vakhrushev SY, Zamfir A, Peter-Katalinić J (2004) 0,2An cross-ring cleavage as a general diagnostic tool for glycan assignment in glycoconjugate mixtures. J Am Soc Mass Spectrom 15:1863–1868
Fong B, Norris C, Lowe E, McJarrow P (2009) Liquid chromatography–high-resolution mass spectrometry for quantitative analysis of gangliosides. Lipids 44:867–874
Svennerholm L, Fredman P (1980) A procedure for the quantitative isolation of brain gangliosides. Biochim Biophys Acta 617:97–109
Vukelić Ž, Metelmann W, Muthing J, Kos M, Peter-Katalinić J (2001) Anencephaly: structural characterization of gangliosides in defined brain regions. Biol Chem 382:259–274
Domon B, Costello CE (1988) A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconj J 5:397–409
Costello CE, Juhasz P, Perreault H (1994) New mass spectral approaches to ganglioside structure determinations. Progr Brain Res 101:45–61
Zamfir A, Lion N, Vukelic Ž, Bindila L, Rossier JS, Girault HH, Peter-Katalinić J (2005) Thin chip microsprayer system coupled to quadrupole time-of-flight mass spectrometer for glycoconjugate analysis. Lab Chip 5:298–307
Bartik P, Maglott A, Entlicher G, Vestweber D, Takeda K, Martin S, Dontenwill M (2008) Detection of a hypersialylated β1 integrin endogenously expressed in the human astrocytoma cell line A172. Int J Oncol 32:1021–1031
Zamfir A, Vukelić Ž, Bindila L, Peter-Katalinić J, Almeida R, Sterling A, Allen M (2004) Fully-automated chip-based nanoelectrospray tandem mass spectrometry of gangliosides from human cerebellum. J Am Soc Mass Spectrom 15:1649–1657
Acknowledgments
This work was supported by the Romanian National Authority for Scientific Research, CNCS-UEFISCDI, projects PN-II-ID-PCE-2011-3-0047, PN-II-PCCA-2011-142, PN-II-RU-TE-2011-2-0008 and FP7 Marie Curie-PIRSES-GA-2010-269256.
Author information
Authors and Affiliations
Corresponding authors
Rights and permissions
About this article
Cite this article
Zamfir, A.D., Fabris, D., Capitan, F. et al. Profiling and sequence analysis of gangliosides in human astrocytoma by high-resolution mass spectrometry. Anal Bioanal Chem 405, 7321–7335 (2013). https://doi.org/10.1007/s00216-013-7173-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-013-7173-x