What is Lipidomics?

Lipidomics is the systematic analysis and characterization of lipids and the subfractions that interact with them in a human body, tissue or cell, in order to understand the structure and function of lipids, and to reveal the relationship between lipid metabolism and the development of cells, organs, and organisms, as well as their growth and pathology. It has been widely used in drug discovery and development, as well as in functional genomics, nutrition, and the environment and pathology. Lipidomics is now widely used in such important fields as drug discovery and development, subsector genomics, functional genomics, nutrition, and environment and health.

What is Dynamic Lipidomics?

Dynamic lipidomics, as part of the broader field of lipidomics, dynamic lipidomics analysis focuses on changes in an organism or cellular system in response to stimuli. Dynamic lipidomics differs from static lipid analysis, which provides a snapshot of cellular lipid abundance and diversity at any given point in time under physiological/pathophysiological conditions. This approach can capture lipid synthesis, turnover, and modification longitudinally to interrogate the dynamic processes that underpin lipid cell homeostasis, such as perturbations in their homeostasis or responses to external stimuli, including different disease conditions.

What is the Lipidome?

The lipidome refers to the complete set of lipids present in a biological system. It encompasses a diverse array of lipid classes. Lipids can be classified into various categories from the point of view of chemical structure and biosynthesis, such as fatty acids (FA), glycerolipids (GL), glycerophospholipids (GP), sterol lipids (ST), sphingolipids (SP), pregnenolone lipids (PR), saccharolipids (SL), polyketides (PK), etc.

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Uses of Stable Isotopes in Lipidomics Analysis

Stable isotopes are now an unavoidable part of lipidomics experiments, providing insights into metabolism and turnover. The precise tracking and quantification of metabolic pathways using stable isotopes has fundamentally changed our view of the role lipids play in cells.

Tracking Lipid Synthesis and Turnover

Stable isotopes have been used widely to investigate lipid synthesis and turnover rates. Researchers are able to incorporate different stable isotope-labeled precursors in lipids which then allows for tracing of the incorporation into various lipid classes. This approach allows measurements of de novo lipogenesis, syntheses from 13C labeled glucose or acetate to the synthesis fatty acids and other lipid.

Investigating Lipid Metabolic Pathways

Stable isotopes provide the opportunity to further dissect intricate lipid metabolic pathways. To track how these lipids are metabolized down different biochemistry routes, the researchers use isotopically labeled substrates. This strategy would help to map the lipid biosynthesis, degradation and remodeling pathways.

Analyzing Lipid Modifications and Interactions

Lipid changes and interactions are critical in many biology systems, with stable isotopes a key tool to study them. To study how these modifications are incorporated into lipid molecules, researchers can use isotopically labeled lipids. This aids in the interpretation of lipid changes occurring during multiple physiological and pathological states.

Quantifying Lipid Metabolic Fluxes

The dynamic regulation of lipid metabolism necessitates a method to quantify changes in lipid metabolic fluxes. Stable isotopes offer invaluable tools to measure fluxes through lipid metabolic pathways, conveying information on the speed of synthesis as opposed degradation and turnover.

Types of Stable Isotopes in Lipidomics Analysis

Carbon-13

Carbon-13 (13C) is a stable isotope of carbon with an atomic mass of 13, accounting for approximately 1% of natural carbon. It is frequently used in metabolic studies due to its ability to trace carbon fluxes in various biological molecules. 13C-labeled substrates such as glucose, acetate, and fatty acids are employed to study lipid synthesis and turnover.

Deuterium

Deuterium (2H), also known as heavy hydrogen, has an atomic mass of 2. It is commonly used in metabolic studies to trace hydrogen incorporation into biological molecules. 2H-labeled water (D2O) and fatty acids are used to study lipid synthesis and turnover. For example, D2O can be employed to track the incorporation of deuterium into lipid molecules, allowing researchers to measure lipid synthesis rates and assess metabolic fluxes. 2H-labeled fatty acids can also be used to study the metabolism of specific lipid classes.

Nitrogen-15

Nitrogen-15 (15N) is a stable isotope of nitrogen with an atomic mass of 15. It is used to study nitrogen metabolism and protein synthesis. 5N-labeled amino acids are incorporated into lipids to study nitrogen fluxes and amino acid utilization in lipid biosynthesis.

Oxygen-18

Oxygen-18 (18O) is a stable isotope of oxygen with an atomic mass of 18. It is used to study oxygen metabolism and oxidation processes. 18O-labeled water or molecular oxygen can be used to track oxygen incorporation into lipid molecules. This is particularly useful for studying lipid oxidation and metabolic pathways involving oxygen.

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Advantages of Stable Isotopes in Lipidomics Analysis

High Sensitivity and Specificity

Stable isotopes, such as Carbon-13 (13C) and Deuterium (2H), enable high-resolution tracking of lipid metabolism due to their distinct mass signatures. This high sensitivity allows for the precise measurement of minor changes in lipid concentrations and metabolic fluxes. For example, 13C-labeled glucose can reveal subtle shifts in fatty acid synthesis pathways that might be missed with less sensitive methods. The specificity of stable isotopes also reduces background noise and increases the accuracy of lipid identification, which is crucial for distinguishing between closely related lipid species.

Quantitative Precision

Stable isotopes are used for precise measurements of metabolic flux and lipid concentrations. Labeling substrates and measuring the degree to which they are integrated into lipid molecules allows accurate estimates of the rate of lipid synthesis, turnover, or modification. This can be done by use! %N labeled amino acids to measure nitrogen into phospholipids are illustrated by studies and reveal information about lipid biosynthesis and remodeling pathways.

Versatility Across Lipid Classes

Stable isotopes can be used to study a variety of lipids, including phospholipids, sphingolipids, glycolipids, and neutral lipids. This adaptability allows for an almost complete lipidomics analysis of the different pathways of lipid metabolism. For example, oxygen-18 (18O) labeling has been used to track lipid oxidation and oxygen-dependent metabolic pathways, while 13c tracers can be used to track carbon fluxes in fatty acids and other digestive-related components of lipids.

Ability to Monitor Dynamic Processes

Stable isotope labelling has proven to indispensable for tracing lipid metabolism over time (a method often referred as dynamic lipidomics) host laboratories. By labeling lipid pools with non-radioactive stable isotopes, researchers can track the rates and patterns of synthesis turnover in specific lipids. Such analysis is important to characterize complex biological dynamics in response to perturbations, e.g. diet changes or drug action, as well during the progression of a disease. Such as longitudinal studies employing 13C-labeled substrates to illustrate how lipid profiles have adapted over time in response to metabolic disturbance.