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MALDI Orbitrap Mass Spectrometry Profiling of Dysregulated Sulfoglycosphingolipids in Renal Cell Carcinoma Tissues
Robert Jirásko1, Michal Holčapek2, Maria Khalikova2
1Department of Analytical Chemistry, Faculty of Chemical Technology, University of Pardubice, Studentská 573, 53210, Pardubice, Czech Republic. robert.jirasko@upce.cz.
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.
Insights
Matrix-assisted laser desorption/ionization Orbitrap mass spectrometry identified significant sulfoglycosphingolipid changes in renal cell carcinoma (RCC) tissues. This study details novel lipid alterations in kidney cancer, aiding diagnostic and therapeutic strategies.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Oncology
Background:
- Renal cell carcinoma (RCC) is the most common kidney cancer.
- Understanding molecular differences between tumor and normal kidney tissue is crucial for diagnosis and treatment.
- Sulfoglycosphingolipids are implicated in various cellular processes, but their role in RCC requires further elucidation.
Purpose of the Study:
- To identify and quantify sulfoglycosphingolipid alterations in RCC tissues using MALDI-Orbitrap-MS.
- To develop and validate a robust MALDI-MS method for clinical sample analysis.
- To differentiate tumor from normal kidney tissue based on lipid profiles.
Main Methods:
- Matrix-assisted laser desorption/ionization coupled with Orbitrap mass spectrometry (MALDI-Orbitrap-MS) was employed.
- High mass accuracy full scan and tandem mass spectrometry were used for sulfoglycosphingolipid identification.
- Multivariate data analysis (MDA), including PCA and OLS-DA, was applied to analyze lipidomic data from 158 tissues of 80 patients.
Main Results:
- Over 120 sulfoglycosphingolipids, many novel, were identified in human RCC samples.
- Significant differences in sulfoglycosphingolipid abundances were observed between tumor and autologous normal kidney tissues.
- MDA successfully visualized distinctions between normal and tumor tissues, highlighting up- and downregulated lipids.
Conclusions:
- MALDI-Orbitrap-MS is a powerful tool for profiling sulfoglycosphingolipids in RCC.
- Specific sulfoglycosphingolipid alterations are characteristic of RCC, offering potential biomarkers.
- The study provides a validated method for semiquantitative analysis and discusses critical aspects of MALDI-MS for clinical applications.