In vitro generation of oxidized standards for lipidomics. Application to major membrane lipid components

Barker-Tejeda, Tomás Clive; Villaseñor, Alma; González-Riaño, Carolina; López-López, Ángeles; Gradillas, Ana; Barbas, Coral

doi:10.1016/j.chroma.2021.462254

Cited by 9 publications

(4 citation statements)

References 77 publications

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“…Surprisingly, all metabolites also presented a matrix effect. This behavior is usually overcome by employing at least one stable isotope-labeled internal standard for each metabolite class [ 30 ], although often these are not available. We, therefore, propose a new approach for metabolite quantification based on the preparation of external calibration curves using the extracted sample.…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Targeted Metabolomic LC-QqQ-MS Method in Allergic Inflammation

et al. 2022

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The transition from mild to severe allergic phenotypes is still poorly understood and there is an urgent need of incorporating new therapies, accompanied by personalized diagnosis approaches. This work presents the development of a novel targeted metabolomic methodology for the analysis of 36 metabolites related to allergic inflammation, including mostly sphingolipids, lysophospholipids, amino acids, and those of energy metabolism previously identified in non-targeted studies. The methodology consisted of two complementary chromatography methods, HILIC and reversed-phase. These were developed using liquid chromatography, coupled to triple quadrupole mass spectrometry (LC-QqQ-MS) in dynamic multiple reaction monitoring (dMRM) acquisition mode and were validated using ICH guidelines. Serum samples from two clinical models of allergic asthma patients were used for method application, which were as follows: (1) corticosteroid-controlled (ICS, n = 6) versus uncontrolled (UC, n = 4) patients, and immunotherapy-controlled (IT, n = 23) versus biologicals-controlled (BIO, n = 12) patients. The results showed significant differences mainly in lysophospholipids using univariate analyses in both models. Multivariate analysis for model 1 was able to distinguish both groups, while for model 2, the results showed the correct classification of all BIO samples within their group. Thus, this methodology can be of great importance for further understanding the role of these metabolites in allergic diseases as potential biomarkers for disease severity and for predicting patient treatment response.

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Section: Discussionmentioning

confidence: 99%

Development of a Novel Targeted Metabolomic LC-QqQ-MS Method in Allergic Inflammation

et al. 2022

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“…Regarding the coverage of metabolites, great efforts have been made to create robust methodologies to cover polar and lipid metabolites independently. In this sense, while analysis of TCA and glycolysis metabolites are a key part of the cellular metabolism, lipids (apart from other roles) are critical regulators of inflammation ( 255 – 257 ). Different studies have demonstrated the importance of lipid mediators in the development of allergy and other related diseases ( 258 ).…”

Section: Discussionmentioning

confidence: 99%

The Importance of Metabolism for Immune Homeostasis in Allergic Diseases

et al. 2021

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There is increasing evidence that the metabolic status of T cells and macrophages is associated with severe phenotypes of chronic inflammation, including allergic inflammation. Metabolic changes in immune cells have a crucial role in their inflammatory or regulatory responses. This notion is reinforced by metabolic diseases influencing global energy metabolism, such as diabetes or obesity, which are known risk factors of severity in inflammatory conditions, due to the metabolic-associated inflammation present in these patients. Since several metabolic pathways are closely tied to T cell and macrophage differentiation, a better understanding of metabolic alterations in immune disorders could help to restore and modulate immune cell functions. This link between energy metabolism and inflammation can be studied employing animal, human or cellular models. Analytical approaches rank from classic immunological studies to integrated analysis of metabolomics, transcriptomics, and proteomics. This review summarizes the main metabolic pathways of the cells involved in the allergic reaction with a focus on T cells and macrophages and describes different models and platforms of analysis used to study the immune system and its relationship with metabolism.

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“…To prevent metabolite oxidation/degradation, sample treatment and exosome isolation were always performed at a temperature of 4°C, and as soon as the exosomes were isolated, these samples were cooled in liquid nitrogen (87,88). To ensure ceramide stability, a recent review reported that ceramides can be oxidized only under very forced conditions such as up to 7 days of oxidation where mainly keto, hydroxy, and hydroperoxyl derivatives were observed (87).…”

Section: Lipid Extractionmentioning

confidence: 99%

Ceramide Composition in Exosomes for Characterization of Glioblastoma Stem-Like Cell Phenotypes

et al. 2022

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Glioblastoma (GBM) is one of the most malignant central nervous system tumor types. Comparative analysis of GBM tissues has rendered four major molecular subtypes. From them, two molecular subtypes are mainly found in their glioblastoma cancer stem-like cells (GSCs) derived in vitro: proneural (PN) and mesenchymal (MES) with nodular (MES-N) and semi-nodular (MES-SN) disseminations, which exhibit different metabolic, growth, and malignancy properties. Many studies suggest that cancer cells communicate between them, and the surrounding microenvironment, via exosomes. Identifying molecular markers that allow the specific isolation of GSC-derived exosomes is key in the development of new therapies. However, the differential exosome composition produced by main GSCs remains unknown. The aim of this study was to determine ceramide (Cer) composition, one of the critical lipids in both cells and their cell-derived exosomes, from the main three GSC phenotypes using mass spectrometry-based lipidomics. GSCs from human tissue samples and their cell-derived exosomes were measured using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) in an untargeted analysis. Complete characterization of the ceramide profile, in both cells and cell-derived exosomes from GSC phenotypes, showed differential distributions among them. Results indicate that such differences of ceramide are chain-length dependent. Significant changes for the C16 Cer and C24:1 Cer and their ratio were observed among GSC phenotypes, being different for cells and their cell-derived exosomes.

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In vitro generation of oxidized standards for lipidomics. Application to major membrane lipid components

Cited by 9 publications

References 77 publications

Development of a Novel Targeted Metabolomic LC-QqQ-MS Method in Allergic Inflammation

Development of a Novel Targeted Metabolomic LC-QqQ-MS Method in Allergic Inflammation

The Importance of Metabolism for Immune Homeostasis in Allergic Diseases

Ceramide Composition in Exosomes for Characterization of Glioblastoma Stem-Like Cell Phenotypes

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