HILIC/ESI-MS determination of gangliosides and other polar lipid classes in renal cell carcinoma and surrounding normal tissues

Abstract

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.

Kidney Lipidomics by Mass Spectrometry Imaging: A Focus on the Glomerulus

“…Moreover, MALDI-MSI was used in parallel to validate the results obtained by HILIC/ESI-MS when comparing patient tumor material with adjacent normal tissue. By targeting polar lipids, such as gangliosides and sulfoglycosphingolipids, and acidic phospholipids [37], MSI results confirmed the findings by HILIC/ESI-MS, consisting of increased phosphatidylinositols, PI 40:5 and PI 40:4, and gangliosides, GM3 34:1, and GM3 42:2, in carcinoma. In another study comparing normal and tumor tissue, MALDI was coupled to an Orbitrap analyzer.…”

Nuclear magnetic resonance and surface-assisted laser desorption/ionization mass spectrometry-based serum metabolomics of kidney cancer

“…They revealed that cluster of 7 metabolites (alanine, creatine, choline, isoleucine, lactate, leucine, and valine) can be used for the early diagnosis of RCC [ 11 ]. Moreover, studies focusing on the metabolite profiling of tissues [ 12 , 22 , 23 ] and urine [ 24 , 25 ] from patients with kidney cancer have been reported. Recently, comprehensive review regarding metabolic profiling of samples from RCC patients has been published [ 19 ].…”

Lipidomic analysis of archival pathology specimen identifies altered lipid signatures in ovarian clear cell carcinoma

“…Glycosphingolipids such as trihexosylceramides (including globotriaosylceramide, Gb3) were elevated in OCCC. These lipids are involved in the metastasis and aggressiveness in cancer [18][19][20][21] The general trend in neutral lipids such as diacylglycerol, triacylglycerol and cholesteryl esters was either no change or a marginal decrease (CE 18:2, CE 20:5) in cancer. Glycerophospholipids along with sphingomyelin are the main components of plasma membranes and constitute the bulk of lipids in tissues [22].…”