Pancreatic ductal adenocarcinoma (PDAC) is one of the most common causes of cancer-related deaths worldwide, accounting for 90% of primary pancreatic tumors with an average 5-year survival rate of less than 10%. PDAC exhibits aggressive biology, which, together with late detection, results in most PDAC patients presenting with unresectable, locally advanced, or metastatic disease. In-depth lipid profiling and screening of potential biomarkers currently appear to be a promising approach for early detection of PDAC or other cancers. Here, we isolated and characterized complex glycosphingolipids (GSL) from normal and tumor pancreatic tissues of patients with PDAC using a combination of TLC, chemical staining, carbohydrate-recognized ligand-binding assay, and LC/ESI-MS2. The major neutral GSL identified were GSL with the terminal blood groups A, B, H, Lea, Leb, Lex, Ley, P1, and PX2 determinants together with globo- (Gb3 and Gb4) and neolacto-series GSL (nLc4 and nLc6). We also revealed that the neutral GSL profiles and their relative amounts differ between normal and tumor tissues. Additionally, the normal and tumor pancreatic tissues differ in type 1/2 core chains. Sulfatides and GM3 gangliosides were the predominant acidic GSL along with the minor sialyl-nLc4/nLc6 and sialyl-Lea/Lex. The comprehensive analysis of GSL in human PDAC tissues extends the GSL coverage and provides an important platform for further studies of GSL alterations; therefore, it could contribute to the development of new biomarkers and therapeutic approaches.
- MeSH
- chromatografie kapalinová MeSH
- chromatografie na tenké vrstvě MeSH
- duktální karcinom slinivky břišní diagnóza patofyziologie MeSH
- gangliosidy chemie MeSH
- glykosfingolipidy * analýza chemie MeSH
- lidé MeSH
- nádorové biomarkery metabolismus MeSH
- nádory slinivky břišní * diagnóza patofyziologie MeSH
- sulfoglykosfingolipidy chemie MeSH
- tandemová hmotnostní spektrometrie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Gangliosides are glycosphingolipids consisting of a ceramide base and a bulky sugar chain that contains one or more sialic acids. This unique structure endows gangliosides with a strong tendency to self-aggregate in solution, as well as in cellular membranes, where they can form nanoscopic assemblies called ganglioside nanodomains. As gangliosides are important biological molecules involved in a number of physiological processes, characterization of their lateral organization in membranes is essential. This review aims at providing comprehensive information about the nanoscale organization of gangliosides in various synthetic models. To this end, the impact of the hydrophobic backbone and the headgroup on the segregation of gangliosides into nanodomains are discussed in detail, as well as the way in which the properties of nanodomains are affected by ligand binding. Small size makes the characterization of ganglioside nanodomains challenging, and we thus highlight the biophysical methods that have advanced this research, such as Monte Carlo Förster resonance energy transfer, atomic force microscopy and approaches based on molecular diffusion.
- MeSH
- buněčná membrána chemie MeSH
- gangliosidy chemie MeSH
- hydrofobní a hydrofilní interakce MeSH
- lidé MeSH
- ligandy MeSH
- lipidové dvojvrstvy chemie MeSH
- mikroskopie atomárních sil MeSH
- rezonanční přenos fluorescenční energie MeSH
- sacharidové sekvence MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Negative-ion hydrophilic liquid chromatography-electrospray ionization mass spectrometry (HILIC/ESI-MS) method has been optimized for the quantitative analysis of ganglioside (GM3) and other polar lipid classes, such as sulfohexosylceramides (SulfoHexCer), sulfodihexosylceramides (SulfoHex2Cer), phosphatidylglycerols (PG), phosphatidylinositols (PI), lysophosphatidylinositols (LPI), and phosphatidylserines (PS). The method is fully validated for the quantitation of the studied lipids in kidney normal and tumor tissues of renal cell carcinoma (RCC) patients based on the lipid class separation and the coelution of lipid class internal standard with the species from the same lipid class. The raw data are semi-automatically processed using our software LipidQuant and statistically evaluated using multivariate data analysis (MDA) methods, which allows the complete differentiation of both groups with 100% specificity and sensitivity. In total, 21 GM3, 28 SulfoHexCer, 26 SulfoHex2Cer, 10 PG, 19 PI, 4 LPI, and 7 PS are determined in the aqueous phase of lipidomic extracts from kidney tumor tissue samples and surrounding normal tissue samples of 20 RCC patients. S-plots allow the identification of most upregulated (PI 40:5, PI 40:4, GM3 34:1, and GM3 42:2) and most downregulated (PI 32:0, PI 34:0, PS 36:4, and LPI 16:0) lipids, which are primarily responsible for the differentiation of tumor and normal groups. Another confirmation of most dysregulated lipids is performed by the calculation of fold changes together with T and p values to highlight their statistical significance. The comparison of HILIC/ESI-MS data and matrix-assisted laser desorption/ionization mass spectrometric imaging (MALDI-MSI) data confirms that lipid dysregulation patterns are similar for both methods. Graphical abstract ᅟ.
The migration potency of differentiated (H-4-II-E, H-4-II-E-C3) and dedifferentiated (H-5) cell lines originated from Reuber hepatoma H 35, and their connection to the morphology and expression of gangliosides were examined. The migration capacity tested by a Transwell assay was fifteen times higher in H-5 cells compared to H-4-II-E cells. The ganglioside pattern was assessed by thin-layer chromatography (TLC). H-4-II-E and H-4-II-E-C3 cell lines expressed Fuc-GM1, which was not found in H-5 cells, whereas H-5 expressed GM3, which was absent in differentiated cell lines. GM3 ganglioside is thought to be one of the key molecules involved in signal transduction of mammalian cells. We conclude that changes in the migration capacity of various hepatoma cell lines might relate to their ganglioside spectra.
- MeSH
- chromatografie na tenké vrstvě metody využití MeSH
- experimentální nádory jater MeSH
- G(M3) gangliosid chemie MeSH
- gangliosidy chemie MeSH
- krysa rodu rattus anatomie a histologie MeSH
- nádorové buněčné linie cytologie MeSH
- pohyb buněk fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus anatomie a histologie MeSH
- zvířata MeSH
Autoimunitní neuropatie jsou imunopatologické stavy, které mohou být provázeny přítomností protilátek proti glykokonjugátům buněčných membrán nervových buněk. Nejvíce se uplatňují autoagresivní reakce proti gangliosidům a některým glykoproteinům, například proti glykoproteinu asociovanému s myelinem. U řady zánětlivých neuropatií lze definovat poměrně přesný profil anti-glykokonjugátových autoprotilátek. V současné době je v klinické laboratorní diagnostice dostupné testování protilátek proti širokému spektru gangliosidů a glykoproteinů, technicky se uplatňují především blotové metody, enzymová imunoanalýza a nepřímá imunofluorescence. Jde o metody časově a finančně dostupné, které mohou přispět ke stanovení či upřesnění diagnózy neuropatie.
- MeSH
- autoimunitní nemoci diagnóza MeSH
- autoprotilátky krev MeSH
- fluorescenční protilátková technika nepřímá metody MeSH
- gangliosidy chemie imunologie MeSH
- glykokonjugáty klasifikace MeSH
- glykoprotein asociovaný s myelinem imunologie MeSH
- imunoblotting metody MeSH
- imunoenzymatické techniky metody MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
