Green light for lipid fingerprinting

The use of targeted lipidomic approaches for the analysis of plant lipids has steadily increased during recent years. We review recent developments of these methods and suggest the introduction of discovery lipidomics as additional approach through a new workflow, lipid fingerprinting, that integrates the advantages of shotgun lipidomics (quantitative data) with LC-MS-based strategies (higher resolution and/or coverage). This article is part of a Special Issue entitled:BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

Is the clinical lipidomics a potential goldmine?

Clinical lipidomics is a new extension of lipidomics to study lipid profiles, pathways, and networks by characterizing and quantifying the complete lipid molecules in cells, biopsy, or body fluids of patients. It undoubtfully has more values if lipidomics can be integrated with the data of clinical proteomic, genomic, and phenomic profiles. A number of challenges, e.g., instability, specificity, and sensitivity, in lipidomics have to be faced and overcome before clinical application. The association of lipidomics data with gene expression and sequencing of lipid-specific proteins/enzymes should be furthermore clarified. Therefore, clinical lipidomics is expected to be more stable during handling, sensitive in response to changes, specific for diseases, efficient in data analyses, and standardized in measurements, in order to meet clinical needs. Clinical lipidomics will become a more important approach in clinical applications and will be the part of “natural” measures for early diagnosis and progress of disease. Thus, clinical lipidomics will be one of the most powerful approaches for disease-specific diagnosis and therapy, once the mystery of lipidomic profiles and metabolic enzymes is deciphered.     

脂质组学研究方法及其应用

脂质不仅是生物膜的骨架成分和能量贮存物质, 越来越多的证据表明, 脂质也参与细胞的许多重要功能。脂质组学是代谢组学的一个重要分支, 主要研究生物体内所有的脂质分子的特性以及它们在蛋白质表达和基因调控过程中的作用。脂质组学是依赖技术驱动的科学。近年来, 随着人们对脂质研究的重视, 脂质组学研究方法和策略有了突破性进展, 在动物上开发出的脂质组学分析方法已经扩展应用到植物上。该文重点介绍脂质组学的研究方法及其应用, 以期推动脂质组学,特别是植物脂质组学的进一步发展。     

Reporting of lipidomics data should be standardized

This article highlights, to our opinion, some of the most pertinent issues related to producing high quality lipidomics data. These issues include pitfalls related to sample collection and storage, lipid extraction, the use of shotgun and LC-MS-based lipidomics approaches, and the identification, annotation and quantification of lipid species. We hope that highlighting these issues will help stimulate efforts to implement reporting standards for dissemination of lipidomics data. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

An enhanced plant lipidomics method based on multiplexed liquid chromatography–mass spectrometry reveals additional insights into cold‐ and drought‐induced membrane remodeling

Within the lipidome of plants a few bulk molecular species hamper the detection of the rest, which are present at relatively low levels. In addition, low‐abundance species are often masked by numerous isobaric interferences, such as those caused by isoelemental species and isotopologues. This scenario not only means that minor species are underrepresented, but also leads to potential misidentifications and limits the structural information gathered by lipidomics approaches. In order to overcome these limitations we have developed a multiplexed liquid chromatography–mass spectrometry lipidomics platform able to achieve an enhanced coverage of plant lipidomes. The platform is based on a single extraction step followed by a series of ultra‐performance liquid chromatography separations. Post‐column flow is then directed to both a triple quadrupole analyzer for targeted profiling and a time‐of‐flight analyzer for accurate mass analysis. As a proof of concept, plants were subjected to cold or drought, which are known to trigger widespread remodeling events in plant cell membranes. Analysis of the leaf lipidome yielded 393 molecular species within 23 different lipid classes. This enhanced coverage allowed us to identify lipid molecular species and even classes that are altered upon stress, allowing hypotheses on role of glycosylinositolphosphoceramides (GIPC), steryl glycosides (SG) and acylated steryl glycosides (ASG) in drought stress to be addressed and confirming the findings from numerous previous studies with a single, wide‐ranging lipidomics approach. This extended our knowledge on membrane remodeling during the drought response, integrating sphingolipids and sterol lipids into the current glycerolipid‐based model.     

Strategies to Improve/Eliminate the Limitations in Shotgun Lipidomics

Direct infusion‐based shotgun lipidomics is one of the most powerful and useful tools in comprehensive analysis of lipid species from lipid extracts of various biological samples with high accuracy/precision. However, despite many advantages, the classical shotgun lipidomics suffers some general dogmas of limitations, such as ion suppression, ambiguous identification of isobaric/isomeric lipid species, and ion source–generated artifacts, restraining the applications in analysis of low‐abundance lipid species, particularly those less ionizable or isomers that yield almost identical fragmentation patterns. This article reviews the strategies (such as modifier addition, prefractionation, chemical derivatization, charge feature utilization) that have been employed to improve/eliminate these limitations in modern shotgun lipidomics approaches (e.g., high mass resolution mass spectrometry–based and multidimensional mass spectrometry–based shotgun lipidomics). Therefore, with the enhancement of these strategies for shotgun lipidomics, comprehensive analysis of lipid species including isomeric/isobaric species is achieved in a more accurate and effective manner, greatly substantiating the aberrant lipid metabolism, signaling trafficking, and homeostasis under pathological conditions.     

Novel advances in shotgun lipidomics for biology and medicine

The field of lipidomics, as coined in 2003, has made profound advances and been rapidly expanded. The mass spectrometry-based strategies of this analytical methodology-oriented research discipline for lipid analysis are largely fallen into three categories: direct infusion-based shotgun lipidomics, liquid chromatography-mass spectrometry-based platforms, and matrix-assisted laser desorption/ionization mass spectrometry-based approaches (particularly in imagining lipid distribution in tissues or cells). This review focuses on shotgun lipidomics. After briefly introducing its fundamentals, the major materials of this article cover its recent advances. These include the novel methods of lipid extraction, novel shotgun lipidomics strategies for identification and quantification of previously hardly accessible lipid classes and molecular species including isomers, and novel tools for processing and interpretation of lipidomics data. Representative applications of advanced shotgun lipidomics for biological and biomedical research are also presented in this review. We believe that with these novel advances in shotgun lipidomics, this approach for lipid analysis should become more comprehensive and high throughput, thereby greatly accelerating the lipidomics field to substantiate the aberrant lipid metabolism, signaling, trafficking, and homeostasis under pathological conditions and their underpinning biochemical mechanisms.     

The evolution of lipidomics through space and time

Although the foundations of mass spectrometry-based lipidomics have been practiced for over 30 years, recent technological advances in ionization modalities in conjunction with robust increases in mass accuracy and resolution have greatly accelerated the emergence, growth and importance of the field of lipidomics. Moreover, advances in the separation sciences, bioinformatic strategies and the availability of robust databases have been synergistically integrated into modern lipidomic technologies leading to unprecedented improvements in the depth, penetrance and precision of lipidomic analyses and identification of their biological and mechanistic significance. The purpose of this "opinion" article is to briefly review the evolution of lipidomics, critique the platforms that have evolved and identify areas that are likely to emerge in the years to come. Through seamlessly integrating a rich repertoire of mass spectrometric, chemical and bioinformatic strategies, the chemical identities and quantities of tens of thousands to hundreds of thousands of different lipid molecular species and their metabolic alterations during physiologic or pathophysiologic perturbations can be obtained. Thus, the field of lipidomics which already has a distinguished history of exciting new discoveries in many disease states holds unparalleled potential to identify the pleiotropic roles of lipids in health and disease at the chemical level. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

Lipidomics in biomedical research-practical considerations

Lipids have many central physiological roles including as structural components of cell membranes, energy storage sources and intermediates in signaling pathways. Lipid-related disturbances are known to underlie many diseases and their co-morbidities. The emergence of lipidomics has empowered researchers to study lipid metabolism at the cellular as well as physiological levels at a greater depth than was previously possible. The key challenges ahead in the field of lipidomics in medical research lie in the development of experimental protocols and in silico techniques needed to study lipidomes at the systems level. Clinical questions where lipidomics may have an impact in healthcare settings also need to be identified, both from the health outcomes and health economics perspectives. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

基于电喷雾电离串联质谱技术的植物脂质组学研究进展

研究表明,脂质不但参与植物的信号转导、小泡运输、细胞骨架重组等多种细胞过程,而且在植物的生长发育和胁迫反应中具有重要作用．但是脂质本身的多样性、复杂性、以及分析手段的滞后限制了人们对脂质的深入认识．电喷雾电离串联质谱(ESI-MS/MS)技术作为一种直接进样的高通量分析技术,能够在短时间内对大多数脂质的不同分子种进行定量分析,极大地方便了人们了解植物因环境变化和生长发育引起的组织内脂质分子种的微量变化．近年来,该技术在植物上的成功应用,推动植物脂质组学研究取得了重要进展,揭示出脂质在植物的逆境胁迫反应、防御反应中的多种功能,促进了植物脂质代谢相关基因的鉴定．而且,该技术与其他脂质分析技术结合,促使人们在脂质的分布、运输、转化和新脂质种类的鉴定方面有新的进展．概要介绍了ESI-MS/MS技术的特点,重点综述了该技术在植物脂质组学研究中的应用进展,并展望了该技术今后的发展方向.     

Lipidomics in tissues, cells and subcellular compartments

Mass spectrometry (MS) advances in recent years have revolutionized the biochemical analysis of lipids in plants, and made possible new theories about the structural diversity and functional complexity of lipids in plant cells. Approaches have been developed to profile the lipidome of plants with increasing chemical and spatial resolution. Here we highlight a variety of methods for lipidomics analysis at the tissue, cellular and subcellular levels. These procedures allow the simultaneous identification and quantification of hundreds of lipids species in tissue extracts by direct-infusion MS, localization of lipids in tissues and cells by laser desorption/ionization MS, and even profiling of lipids in individual subcellular compartments by direct-organelle MS. Applications of these approaches to achieve improved understanding of plant lipid metabolism, compartmentation and function are discussed.     

鸟枪脂组学研究进展及其应用

继基因组学之后,针对各种代谢物的组学研究蓬勃兴起,鸟枪脂组学(shotgun lipidom ics)作为脂类研究的重要新兴手段,在创立和初期发展的过程中便已经展示出惊人的潜力,随着相关技术的进一步完善和发展,必将成为系统生物学的组成部分,在生物医学的研究和应用中发挥难以替代的重要作用。鸟枪脂组学利用质谱技术对全部或单一脂类及其相关分子进行系统分析,研究其改变对生物体所产生的作用并探讨其作用机制。传统脂类分析中的瓶颈问题在以电喷射离子质谱为基础的脂组学方法出现后获得了突破,使脂类分析进入高通量、高精度和高效能的时代。脂类在生物体内分布广泛、种类众多,并且与人类疾病密切相关。将脂组学分析方法运用于疾病相关的特异脂类标志物的发现并揭示其在疾病发生发展等复杂过程中的作用,可能为疾病的诊断治疗提供新的思路和策略。     

Spatial mapping of lipids at cellular resolution in embryos of cotton

Advances in mass spectrometry (MS) have made comprehensive lipidomics analysis of complex tissues relatively commonplace. These compositional analyses, although able to resolve hundreds of molecular species of lipids in single extracts, lose the original cellular context from which these lipids are derived. Recently, high-resolution MS of individual lipid droplets from seed tissues indicated organelle-to-organelle variation in lipid composition, suggesting that heterogeneity of lipid distributions at the cellular level may be prevalent. Here, we employed matrix-assisted laser desorption/ionization-MS imaging (MALDI-MSI) approaches to visualize lipid species directly in seed tissues of upland cotton (Gossypium hirsutum). MS imaging of cryosections of mature cotton embryos revealed a distinct, heterogeneous distribution of molecular species of triacylglycerols and phosphatidylcholines, the major storage and membrane lipid classes in cotton embryos. Other lipids were imaged, including phosphatidylethanolamines, phosphatidic acids, sterols, and gossypol, indicating the broad range of metabolites and applications for this chemical visualization approach. We conclude that comprehensive lipidomics images generated by MALDI-MSI report accurate, relative amounts of lipid species in plant tissues and reveal previously unseen differences in spatial distributions providing for a new level of understanding in cellular biochemistry.     

Glycerolipid and cholesterol ester analyses in biological samples by mass spectrometry

Neutral lipids are a diverse family of hydrophobic biomolecules that have important roles in cellular biochemistry of all living species but have in common the property of charge neutrality. A large component of neutral lipids is the glycerolipids composed of triacylglycerols, diacylglycerols, and monoacylglycerols that can serve as cellular energy stores as well as signaling molecules. Another abundant lipid class in many cells is the cholesterol esters that are on one hand sterols and the other fatty acyl lipids, but in either case are neutral lipids involved in cholesterol homeostasis and transport in the blood. The analysis of these molecules in the context of lipidomics remains challenging because of their charge neutrality and the complex mixtures of molecular species present in cells. Various techniques have been used to ionize these neutral lipids prior to mass spectrometric analysis including electron ionization, atmospheric chemical ionization, electrospray ionization and matrix assisted laser desorption/ionization. Various approaches to deal with the complex mixture of molecular species have been developed including shotgun lipidomics and chromatographic-based separations such as gas chromatography, reversed phase liquid chromatography, and normal phase liquid chromatography. Several applications of these approaches are discussed. .     

Multi-OMICS: a critical technical perspective on integrative lipidomics approaches

During the past decades, high-throughput approaches for analyzing different molecular classes such as nucleic acids, proteins, metabolites, and lipids have grown rapidly. These approaches became powerful tools for getting a fundamental understanding of biological systems. Considering each approach and its results separately, relations and causal connections between these classes have no chance to be revealed, since only separate molecular snapshots are provided. Only a combined approach, not fully established yet, with the integration of the corresponding data, might yield a comprehensive and complete understanding of biological processes, such as crosstalk and interactions in signaling pathways. Taking two or more omics-methods into consideration for analysis is referred to as a multi-omics approach, which is gradually evolving. In this critical note, we briefly discuss the relevance, challenges, current state, and potential of data integration from multi-omics approaches, with a special focus on lipidomics analysis, listing the advantages and gaps in this field. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

Lipidomics,en route to accurate quantitation

Accurate quantitation is prerequisite for the sustainable development of lipidomics via enabling its applications in various biological and biomedical settings. In this review, the technical considerations and limitations of existent lipidomics technologies, particularly in terms of accurate quantitation; as well as the potential sources of errors along a typical lipidomic workflow that could ultimately give rise to quantitative inaccuracies will be addressed. Furthermore, the pressing need to exercise stricter definitions of terms and protocol standardization pertaining to quantitative lipidomics will be critically discussed, as quantitative accuracy may substantially impact upon the persevering development of lipidomics in the long run. This article is part of a Special Issue entitled: BBALIP_Lipidomics Opinion Articles edited by Sepp Kohlwein.     

脂质组学分析方法进展及其在癌症研究中的应用

脂质占人体内源性代谢物的一半以上,种类繁多,结构复杂,因而具有多种生物功能,与多种生命活动密切相关。脂质组学是代谢组学分支的新兴学科,它可以通过比较不同生理状态下脂质含量的变化,寻找代谢通路中关键的脂质生物标志物,最终揭示脂质在各种生命活动中的作用机制。随着质谱技术的进步,脂质组学在疾病脂类生物标志物的识别、疾病诊断、药物作用机制的研究等方面已展现出广泛的应用前景。本文主要就脂质组学近几年的分析方法进展及其在癌症中的最新应用进行了综述。     

Lipidomics: new tools and applications

Once viewed simply as a reservoir for carbon storage, lipids are no longer cast as bystanders in the drama of biological systems. The emerging field of lipidomics is driven by technology, most notably mass spectrometry, but also by complementary approaches for the detection and characterization of lipids and their biosynthetic enzymes in living cells. The development of these integrated tools promises to greatly advance our understanding of the diverse biological roles of lipids.     

Advances in chemical proteomic evaluation of lipid kinases—DAG kinases as a case study

Advancements in chemical proteomics and mass spectrometry lipidomics are providing new opportunities to understand lipid kinase activity, specificity, and regulation on a global cellular scale. Here, we describe recent developments in chemical biology of lipid kinases with a focus on those members that phosphorylate diacylglycerols. We further discuss future implications of how these mass spectrometry–based approaches can be adapted for studies of additional lipid kinase members with the aim of bridging the gap between protein and lipid kinase–focused investigations.