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Article: Kidney derived apolipoprotein M and its role in acute kidney injury.

Bisgaard, Line S / Christensen, Pernille M / Oh, Jeongah / Torta, Federico / Füchtbauer, Ernst-Martin / Nielsen, Lars Bo / Christoffersen, Christina

Frontiers in pharmacology

2024 Volume 15, Page(s) 1328259

Abstract: Aim: ...

Abstract	Aim:
Language	English
Publishing date	2024-01-19
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587355-6
ISSN	1663-9812
ISSN	1663-9812
DOI	10.3389/fphar.2024.1328259
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Article ; Online: Author Correction: Q-RAI data-independent acquisition for lipidomic quantitative profiling.

Chang, Jing Kai / Teo, Guoshou / Pewzner-Jung, Yael / Cuthbertson, Daniel J / Futerman, Anthony H / Wenk, Markus R / Choi, Hyungwon / Torta, Federico

Scientific reports

2024 Volume 14, Issue 1, Page(s) 4908

Language	English
Publishing date	2024-02-28
Publishing country	England
Document type	Published Erratum
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-55396-9
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Article ; Online: Mass Spectrometry Analysis of the Human Brain Sphingolipidome.

Chua, Xin Ying / Huang, Ryan / Herr, Deron / Lai, Mitchell K P / Wenk, Markus R / Torta, Federico

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2561, Page(s) 233–243

Abstract: In recent decades, mass spectrometry-based lipidomics has provided a fertile environment for scientific investigations of biochemical and mechanistic processes in biological systems. Notably, this approach has been used to characterize physiological and ... ...

Abstract	In recent decades, mass spectrometry-based lipidomics has provided a fertile environment for scientific investigations of biochemical and mechanistic processes in biological systems. Notably, this approach has been used to characterize physiological and pathological processes relevant to the central nervous system by identifying changes in the sphingolipid content in the brain, cerebral spinal fluid, and blood plasma. However, despite a preponderance of studies identifying correlations between specific lipids and disease progression, this powerful resource has not yet substantively translated into clinically useful diagnostic assays. Part of this gap may be explained by insufficient depth of the lipidomic profiles in many studies, by lab-to-lab inconsistencies in methodology, and a lack of absolute quantification. These issues limit the identification of specific molecular species and the harmonization of results across independent studies. In this chapter, we contextualize these issues with recent reports identifying correlations between brain lipids and neurological diseases, and we describe the workflow our group has optimized for analysis of the blood plasma sphingolipidome, adapted to the characterization of the human brain tissue.
MeSH term(s)	Humans ; Mass Spectrometry/methods ; Brain ; Lipidomics ; Sphingolipids ; Workflow
Chemical Substances	Sphingolipids
Language	English
Publishing date	2022-11-18
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2655-9_12
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Article ; Online: Deficiency in the omega-3 lysolipid transporter Mfsd2a leads to aberrant oligodendrocyte lineage development and hypomyelination.

Sengottuvel, Vetrivel / Hota, Monalisa / Oh, Jeongah / Galam, Dwight L / Wong, Bernice H / Wenk, Markus R / Ghosh, Sujoy / Torta, Federico / Silver, David L

The Journal of clinical investigation

2023 Volume 133, Issue 12

Abstract: Patients with autosomal recessive microcephaly 15 caused by deficiency in the sodium-dependent lysophosphatidylcholine (LPC) transporter major facilitator superfamily domain-containing 2a (Mfsd2a) present with both microcephaly and hypomyelination, ... ...

Abstract	Patients with autosomal recessive microcephaly 15 caused by deficiency in the sodium-dependent lysophosphatidylcholine (LPC) transporter major facilitator superfamily domain-containing 2a (Mfsd2a) present with both microcephaly and hypomyelination, suggesting an important role for LPC uptake by oligodendrocytes in the process of myelination. Here we demonstrate that Mfsd2a is specifically expressed in oligodendrocyte precursor cells (OPCs) and is critical for oligodendrocyte development. Single-cell sequencing of the oligodendrocyte lineage revealed that OPCs from OPC-specific Mfsd2a-KO mice (2aOKO mice) underwent precocious differentiation into immature oligodendrocytes and impaired maturation into myelinating oligodendrocytes, correlating with postnatal brain hypomyelination. 2aOKO mice did not exhibit microcephaly, a finding consistent with the notion that microcephaly is the consequence of an absence of LPC uptake at the blood-brain barrier rather than a deficiency in OPCs. Lipidomic analysis showed that OPCs and iOLs from 2aOKO mice had significantly decreased levels of phospholipids containing omega-3 fatty acids, with a corresponding increase in unsaturated fatty acids, the latter being products of de novo synthesis governed by Srebp-1. RNA-Seq indicated activation of the Srebp-1 pathway and defective expression of regulators of oligodendrocyte development. Taken together, these findings indicate that the transport of LPCs by Mfsd2a in OPCs is important for maintaining OPC state to regulate postnatal brain myelination.
MeSH term(s)	Animals ; Mice ; Microcephaly/metabolism ; Sterol Regulatory Element Binding Protein 1/metabolism ; Cell Lineage ; Symporters/metabolism ; Mice, Knockout ; Membrane Transport Proteins/metabolism ; Fatty Acids, Omega-3/metabolism ; Oligodendroglia/metabolism ; Cell Differentiation
Chemical Substances	Sterol Regulatory Element Binding Protein 1 ; Symporters ; Membrane Transport Proteins ; Fatty Acids, Omega-3 ; Mfsd2a protein, mouse
Language	English
Publishing date	2023-06-15
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	3067-3
ISSN	1558-8238 ; 0021-9738
ISSN (online)	1558-8238
ISSN	0021-9738
DOI	10.1172/JCI164118
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Article ; Online: Q-RAI data-independent acquisition for lipidomic quantitative profiling.

Chang, Jing Kai / Teo, Guoshou / Pewzner-Jung, Yael / Cuthbertson, Daniel J / Futerman, Anthony H / Wenk, Markus R / Choi, Hyungwon / Torta, Federico

Scientific reports

2023 Volume 13, Issue 1, Page(s) 19281

Abstract: Untargeted lipidomics has been increasingly adopted for hypothesis generation in a biological context or discovery of disease biomarkers. Most of the current liquid chromatography mass spectrometry (LC-MS) based untargeted methodologies utilize a data ... ...

Abstract	Untargeted lipidomics has been increasingly adopted for hypothesis generation in a biological context or discovery of disease biomarkers. Most of the current liquid chromatography mass spectrometry (LC-MS) based untargeted methodologies utilize a data dependent acquisition (DDA) approach in pooled samples for identification and MS-only acquisition for semi-quantification in individual samples. In this study, we present for the first time an untargeted lipidomic workflow that makes use of the newly implemented Quadrupole Resolved All-Ions (Q-RAI) acquisition function on the Agilent 6546 quadrupole time-of-flight (Q-TOF) mass spectrometer to acquire MS2 spectra in data independent acquisition (DIA) mode. This is followed by data processing and analysis on MetaboKit, a software enabling DDA-based spectral library construction and extraction of MS1 and MS2 peak areas, for reproducible identification and quantification of lipids in DIA analysis. This workflow was tested on lipid extracts from human plasma and showed quantification at MS1 and MS2 levels comparable to multiple reaction monitoring (MRM) targeted analysis of the same samples. Analysis of serum from Ceramide Synthase 2 (CerS2) null mice using the Q-RAI DIA workflow identified 88 lipid species significantly different between CerS2 null and wild type mice, including well-characterized changes previously associated with this phenotype. Our results show the Q-RAI DIA as a reliable option to perform simultaneous identification and reproducible relative quantification of lipids in exploratory biological studies.
MeSH term(s)	Humans ; Animals ; Mice ; Lipidomics/methods ; Mass Spectrometry/methods ; Chromatography, Liquid/methods ; Lipids ; Ions
Chemical Substances	Lipids ; Ions
Language	English
Publishing date	2023-11-07
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-023-46312-8
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Article ; Online: Mapping the distribution of double bond location isomers in lipids across mouse tissues.

Ren, Hanlin / Triebl, Alexander / Muralidharan, Sneha / Wenk, Markus R / Xia, Yu / Torta, Federico

The Analyst

2021 Volume 146, Issue 12, Page(s) 3899–3907

Abstract: Lipids are highly diverse and essential biomolecules in all living systems. As lipid homeostasis is often perturbed in metabolic diseases, these molecules can serve as both biomarkers and drug targets. The development of modern mass spectrometry (MS) ... ...

Abstract	Lipids are highly diverse and essential biomolecules in all living systems. As lipid homeostasis is often perturbed in metabolic diseases, these molecules can serve as both biomarkers and drug targets. The development of modern mass spectrometry (MS) provided the platform for large-scale lipidomic studies at the level of molecular species. Traditionally, more detailed structural information, such as the C[double bond, length as m-dash]C location, was mostly assumed instead of properly measured, though the specific isomers were indicated as potential biomarkers of cancers or cardiovascular diseases. Recent C[double bond, length as m-dash]C localization methods, including the Paternò-Büchi (PB) reaction, have shown the prevalent and heterogeneous distribution of C[double bond, length as m-dash]C location in lipids across tissues. Mapping the lipidome of model animals at the level of C[double bond, length as m-dash]C position would increase the understanding of the metabolism and function of lipid isomers, facilitating clinical research. In this study, we employed an online PB reaction on a liquid chromatography-high resolution MS platform to map C[double bond, length as m-dash]C location isomers in five different murine tissues. We analyzed phosphatidylcholines, phosphatidylethanolamines, and sphingomyelins; we relatively quantified and mapped the distribution of ∼30 groups of co-existing isomers, characterized by different chain lengths and degrees of unsaturation. More specifically, we performed relative quantitation of four isomers of the C16:1 fatty acyl, which included rarely reported n-10 and n-5 species besides n-9 and n-7 isomers. We showed a small variation of the isomers' relative composition among individual animals (<20%) but significant differences across different lipid species and mouse tissues. Our results provided an initial database to map alternative lipid metabolic pathways at the tissue level.
MeSH term(s)	Animals ; Chromatography, Liquid ; Isomerism ; Mass Spectrometry ; Mice ; Sphingomyelins
Chemical Substances	Sphingomyelins
Language	English
Publishing date	2021-06-16
Publishing country	England
Document type	Journal Article
ZDB-ID	210747-8
ISSN	1364-5528 ; 0003-2654
ISSN (online)	1364-5528
ISSN	0003-2654
DOI	10.1039/d1an00449b
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Article ; Online: LICAR: An Application for Isotopic Correction of Targeted Lipidomic Data Acquired with Class-Based Chromatographic Separations Using Multiple Reaction Monitoring.

Gao, Liang / Ji, Shanshan / Burla, Bo / Wenk, Markus R / Torta, Federico / Cazenave-Gassiot, Amaury

Analytical chemistry

2021 Volume 93, Issue 6, Page(s) 3163–3171

Abstract: Lipidomics is developing as an important area in biomedical and clinical research. Reliable quantification of lipid species is required for clinical translation of lipidomic studies. Hydrophilic interaction chromatography (HILIC), normal-phase liquid ... ...

Abstract	Lipidomics is developing as an important area in biomedical and clinical research. Reliable quantification of lipid species is required for clinical translation of lipidomic studies. Hydrophilic interaction chromatography (HILIC), normal-phase liquid chromatography (NPLC), and supercritical fluid chromatography (SFC) are commonly used techniques in lipidomics and provide class-based separation of lipids. While co-elution of lipid species and their internal standards is an advantage for accurate quantification, it leads to isotopic overlap between species of the same lipid class. In shotgun lipidomics, isotopic correction is typically done based on elemental formulas of precursor ions. In multiple reaction monitoring (MRM) analyses, however, this approach should not be used, as the overall contribution of heavy isotopes to the MRM transitions' intensities depends on their location in the molecule with respect to the fragmentation pattern. We present an algorithm, provided in the R programming language, for isotopic correction in class-based separation using MRM, extracting relevant structural information from MRM transitions to apply adequate isotopic correction factors. Using standards, we show that our algorithm accurately estimates the isotopic contribution of isotopologues to MRM transitions' measured intensities. Using human plasma as an example, we demonstrate the necessity of adequate isotopic correction for accurate quantitation of lipids measured by MRM with class-based chromatographic separation. We show that over a third of the measured phosphatidylcholine species had their intensity corrected by more than 10%. This isotopic correction algorithm and R-implemented application enable a more accurate quantification of lipids in class-based separation-MRM, a prerequisite for successful translation of lipidomic applications.
MeSH term(s)	Chromatography, Liquid ; Chromatography, Supercritical Fluid ; Humans ; Hydrophobic and Hydrophilic Interactions ; Lipidomics ; Lipids
Chemical Substances	Lipids
Language	English
Publishing date	2021-02-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.0c04565
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Article ; Online: Sphingolipid metabolism during Toll-like receptor 4 (TLR4)-mediated macrophage activation.

Olona, Antoni / Hateley, Charlotte / Muralidharan, Sneha / Wenk, Markus R / Torta, Federico / Behmoaras, Jacques

British journal of pharmacology

2021 Volume 178, Issue 23, Page(s) 4575–4587

Abstract: Macrophage activation in response to stimulation of Toll-like receptor 4 (TLR4) provides a paradigm for investigating energy metabolism that regulates the inflammatory response. TLR4-mediated pro-inflammatory macrophage activation is characterized by ... ...

Abstract	Macrophage activation in response to stimulation of Toll-like receptor 4 (TLR4) provides a paradigm for investigating energy metabolism that regulates the inflammatory response. TLR4-mediated pro-inflammatory macrophage activation is characterized by increased glycolysis and altered mitochondrial metabolism, supported by selective amino acid uptake and/or usage. Fatty acid metabolism remains as a highly complex rewiring that accompanies classical macrophage activation. TLR4 activation leads to de novo synthesis of fatty acids, which flux into sphingolipids, complex lipids that form the building blocks of eukaryotic cell membranes and regulate cell function. Here, we review the importance of TLR4-mediated de novo synthesis of membrane sphingolipids in macrophages. We first highlight fatty acid metabolism during TLR4-driven macrophage immunometabolism. We then focus on the temporal dynamics of sphingolipid biosynthesis and emphasize the modulatory role of some sphingolipid species (i.e. sphingomyelins, ceramides and glycosphingolipids) on the pro-inflammatory and pro-resolution phases of LPS/TLR4 activation in macrophages.
MeSH term(s)	Lipopolysaccharides/pharmacology ; Macrophage Activation ; Macrophages/metabolism ; Sphingolipids/metabolism ; Toll-Like Receptor 4/metabolism
Chemical Substances	Lipopolysaccharides ; Sphingolipids ; Toll-Like Receptor 4
Language	English
Publishing date	2021-09-02
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	80081-8
ISSN	1476-5381 ; 0007-1188
ISSN (online)	1476-5381
ISSN	0007-1188
DOI	10.1111/bph.15642
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Article ; Online: The lipid transporter Mfsd2a maintains pulmonary surfactant homeostasis.

Wong, Bernice H / Mei, Ding / Chua, Geok Lin / Galam, Dwight L / Wenk, Markus R / Torta, Federico / Silver, David L

The Journal of biological chemistry

2022 Volume 298, Issue 3, Page(s) 101709

Abstract: Pulmonary surfactant is a lipoprotein complex essential for lung function, and insufficiency or altered surfactant composition is associated with major lung diseases, such as acute respiratory distress syndromes, idiopathic pulmonary fibrosis, and ... ...

Abstract	Pulmonary surfactant is a lipoprotein complex essential for lung function, and insufficiency or altered surfactant composition is associated with major lung diseases, such as acute respiratory distress syndromes, idiopathic pulmonary fibrosis, and chronic obstructive pulmonary disease. Pulmonary surfactant is primarily composed of phosphatidylcholine (PC) in complex with specialized surfactant proteins and secreted by alveolar type 2 (AT2) cells. Surfactant homeostasis on the alveolar surface is balanced by the rates of synthesis and secretion with reuptake and recycling by AT2 cells, with some degradation by pulmonary macrophages and loss up the bronchial tree. However, whether phospholipid (PL) transporters exist in AT2 cells to mediate reuptake of surfactant PL remains to be identified. Here, we demonstrate that major facilitator superfamily domain containing 2a (Mfsd2a), a sodium-dependent lysophosphatidylcholine (LPC) transporter, is expressed at the apical surface of AT2 cells. A mouse model with inducible AT2 cell-specific deficiency of Mfsd2a exhibited AT2 cell hypertrophy with reduced total surfactant PL levels because of reductions in the most abundant surfactants, PC containing dipalmitic acid, and PC species containing the omega-3 fatty acid docosahexaenoic acid. These changes in surfactant levels and composition were mirrored by similar changes in the AT2 cell lipidome. Mechanistically, direct tracheal instillation of fluorescent LPC and PC probes indicated that Mfsd2a mediates the uptake of LPC generated by pulmonary phospholipase activity in the alveolar space. These studies reveal that Mfsd2a-mediated LPC uptake is quantitatively important in maintaining surfactant homeostasis and identify this lipid transporter as a physiological component of surfactant recycling.
MeSH term(s)	Animals ; Docosahexaenoic Acids/metabolism ; Homeostasis ; Lung/metabolism ; Lysophosphatidylcholines/metabolism ; Membrane Transport Proteins/metabolism ; Mice ; Phosphatidylcholines ; Phospholipids ; Pulmonary Surfactants ; Symporters/metabolism
Chemical Substances	Lysophosphatidylcholines ; Membrane Transport Proteins ; Mfsd2a protein, mouse ; Phosphatidylcholines ; Phospholipids ; Pulmonary Surfactants ; Symporters ; Docosahexaenoic Acids (25167-62-8)
Language	English
Publishing date	2022-02-10
Publishing country	United States
Document type	Journal Article
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1016/j.jbc.2022.101709
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Article ; Online: Dual mass spectrometry as a tool to improve annotation and quantification in targeted plasma lipidomics.

Gao, Liang / Cazenave-Gassiot, Amaury / Burla, Bo / Wenk, Markus R / Torta, Federico

Metabolomics : Official journal of the Metabolomic Society

2020 Volume 16, Issue 5, Page(s) 53

Abstract: Introduction: High quality data, based on reliable quantification and clear identification of the reported lipid species, are required for the clinical translation of human plasma lipidomic studies.: Objective: Lipid quantification can be efficiently ...

Abstract	Introduction: High quality data, based on reliable quantification and clear identification of the reported lipid species, are required for the clinical translation of human plasma lipidomic studies. Objective: Lipid quantification can be efficiently performed on triple quadrupole (QqQ) mass spectrometers in targeted multiple reaction monitoring (MRM) mode. However, a series of issues can be encountered when aiming at unambiguous identification and accurate quantification, including (i) resolving peaks of polyunsaturated species, (ii) discriminating between plasmanyl-, plasmenyl- and odd chain species and (iii) resolving the isotopic overlap between co-eluting lipid species. Methods: As a practical tool to improve the quality of targeted lipidomics studies, we applied a Dual MS platform by simultaneously coupling a reversed-phase liquid chromatography separation to a QqQ and a quadrupole-time of flight (Q-ToF) mass spectrometers. In one single experiment, this platform allows to correctly identify, by high-resolution MS and MS/MS, the peaks that are quantified by MRM. Results: As proof of concept, we applied the platform on glycerophosphocholines (GPCs) and sphingomyelins (SMs), which are highly abundant in human plasma and play crucial roles in various physiological functions. Our results demonstrated that Dual MS could provide a higher level of confidence in the identification and quantification of GPCs and SMs in human plasma. The same approach can also be applied to improve the study of other lipid classes and expanded for the identification of novel lipid molecular species. Conclusions: This methodology might have a great potential to achieve a better specificity in the quantification of lipids by targeted lipidomics in high-throughput studies.
MeSH term(s)	Chromatography, Liquid ; Humans ; Lipidomics ; Lipids/blood ; Lipids/chemistry ; Molecular Structure ; Tandem Mass Spectrometry
Chemical Substances	Lipids
Language	English
Publishing date	2020-04-17
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2250617-2
ISSN	1573-3890 ; 1573-3882
ISSN (online)	1573-3890
ISSN	1573-3882
DOI	10.1007/s11306-020-01677-z
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