title: MALDI Orbitrap Mass Spectrometry Profiling of Dysregulated Sulfoglycosphingolipids in Renal Cell Carcinoma Tissues.
abstract: Matrix-assisted laser desorption/ionization coupled with Orbitrap mass spectrometry (MALDI-Orbitrap-MS) is used for the clinical study of patients with renal cell carcinoma (RCC), as the most common type of kidney cancer. Significant changes in sulfoglycosphingolipid abundances between tumor and autologous normal kidney tissues are observed. First, sulfoglycosphingolipid species in studied RCC samples are identified using high mass accuracy full scan and tandem mass spectra. Subsequently, optimization, method validation, and statistical evaluation of MALDI-MS data for 158 tissues of 80 patients are discussed. More than 120 sulfoglycosphingolipids containing one to five hexosyl units are identified in human RCC samples based on the systematic study of their fragmentation behavior. Many of them are recorded here for the first time. Multivariate data analysis (MDA) methods, i.e., unsupervised principal component analysis (PCA) and supervised orthogonal partial least square discriminant analysis (OPLS-DA), are used for the visualization of differences between normal and tumor samples to reveal the most up- and downregulated lipids in tumor tissues. Obtained results are closely correlated with MALDI mass spectrometry imaging (MSI) and histologic staining. Important steps of the present MALDI-Orbitrap-MS approach are also discussed, such as the selection of best matrix, correct normalization, validation for semiquantitative study, and problems with possible isobaric interferences on closed masses in full scan mass spectra. Graphical Abstract ᅟ.
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210, Pardubice, Czech Republic. robert.jirasko@upce.cz.
authors.forename: Robert
authors.identifier: 0000-0001-5795-5017
authors.initials: R
authors.lastname: Jirásko
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210, Pardubice, Czech Republic.
authors.forename: Michal
authors.initials: M
authors.lastname: Holčapek
authors.affiliation: Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210, Pardubice, Czech Republic.
authors.forename: Maria
authors.initials: M
authors.lastname: Khalikova
authors.affiliation: Department of Oncology, Faculty of Medicine and Dentistry, Palacký University, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic.
authors.forename: David
authors.initials: D
authors.lastname: Vrána
authors.affiliation: Department of Urology, Faculty of Medicine and Dentistry, Palacký University, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic.
authors.forename: Vladimír
authors.initials: V
authors.lastname: Študent
authors.affiliation: Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacký University, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Prouzová
authors.affiliation: Department of Oncology, Faculty of Medicine and Dentistry, Palacký University, I.P. Pavlova 6, 775 20, Olomouc, Czech Republic.
authors.forename: Bohuslav
authors.initials: B
authors.lastname: Melichar
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis; D002292:Carcinoma, Renal Cell / Q000737:chemistry / Q000175:diagnosis / Q000473:pathology*; D006801:Humans; D007668:Kidney / Q000737:chemistry / Q000473:pathology*; D007680:Kidney Neoplasms / Q000737:chemistry / Q000175:diagnosis / Q000473:pathology*; D016018:Least-Squares Analysis; D015999:Multivariate Analysis; D025341:Principal Component Analysis; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013433:Sulfoglycosphingolipids / Q000032:analysis*
chemical_list: D014408:Biomarkers, Tumor; D013433:Sulfoglycosphingolipids
keywords: MALDI; Orbitrap mass spectrometry; Renal cell carcinoma; Sulfatide; Sulfoglycosphingolipids