Exosome Lipidomics Service

What Is Exosome Lipidomics

Extracellular vesicles (EVs) occur in most bodily fluids and cell culture supernatants. Exosomes, a part of two EV subgroups, are small vesicles (traditionally considered 50-150 nm) originating from an intracellular compartment, called Multivesicular Bodies (MVB, or Late Endosomes) and released out of the cell. Exosomes are involved in cell-to-cell communication through the biological transfer of lipids, proteins, DNAs, RNAs, mRNAs, and miRNAs.

Exosomes have various composition, including proteins, nucleic acids, lipids, and other metabolites. The lipid composition of exosomes resembles lipid rafts, and exosomes have higher lipid content and greater detergent stability compared to other extracellular vesicles. The lipid molecules of exosomal membranes can interact with receptors on recipient cell membranes, such as G-protein coupled receptors, and other lipid molecules, thereby regulating intracellular signaling pathways and biological activities within recipient cells. Lipids play essential roles not only in the structures of exosomal membranes, but also in exosome formation and releasing to the extracellular environment. Knowing the exosome lipid composition is important to understand the biology of vesicles and to investigate possible medical applications. Exosome lipidomics analysis can help us to understand vesicle biogenesis and function, identify lipid-based biomarkers and unravel its monitoring function in diseases.

Figure 1. Lipid composition of exosomes. (PC: phosphatidylcholine; PE: phosphatidylethanolamine; SM: sphingomyelin; CER: ceramide; TG: triacylglyceride; DG: diacylglyceride; CE: cholesteryl ester; S1P: sphingosine-1-phosphate)

What Our Exosomal Lipidomics Service Offers

Lipidomics analysis presents challenges such as high lipid heterogeneity, difficulty in identification, and the need for high sample purity. Exosomal lipidomics analysis faces the common issue of limited sample availability, which poses challenges in achieving accurate qualitative and quantitative results. To deliver high-quality exosomal lipidomics data, we have developed a unique exosome preparation technique to ensure high sample purity before analysis. We help you identify and quantify the lipids in the exosomes quickly, accurately and efficiently. Sophisticated shotgun method and targeted lipidomic assays will be used for in-depth analysis of the exosome lipidomics. We can use powerful facilities to analyze the complexity of lipids, such as Mass spectrometry (MS), high-performance liquid chromatography (HPLC), liquid chromatography coupled MS (LC-MS), nuclear magnetic resonance spectroscopy (NMR), etc.

Additionally, Our comprehensive targeted metabolomics platform covers hundreds of lipid species, ensuring both extensive coverage and high accuracy in qualitative and quantitative lipid identification.

With cutting-edge facilities and optimized protocols, we can provide comprehensive services for exosome research, including:

• Exosome isolation
• Lipidomics analysis
• Data analysis
• A detailed report

Technical workflow

Sample Requirement

We can accept various types of samples, including but not limited:

Deliverables

At Creative Proteomics, we will provide a detailed report to our clients. Our reports contain:

• Experiment procedures and parameters of instruments
• MS data with putative identification based on the m/z ratio of the analytes
• Differential analysis of lipids between treatment groups if needed

Advantages

• One-stop service
• Cutting-edge facilities
• Reliable and reproducible data
• Fast turnaround time

Exosome lipidomics service at Creative Proteomics helps our clients obtain the most information from your exosomes to accelerate basic exosome research, circulating biomarker discovery, or other exosome-related researches. In addition, we have other advanced platforms to isolate, purify, and study exosomes. As every project has different requirements, please contact our specialists to discuss your specific needs. We are looking forward to cooperating with you.

Case 1

Phospholipase D and phosphatidic acid in the biogenesis and cargo loading of extracellular vesicles

Journal: Lipid Res.
Published: 2018

Extracellular vesicles released by live cells, including exosomes and microvesicles, have become crucial cellular components supporting intercellular communication. Due to their potential therapeutic significance, researchers are striving to characterize the contents of these vesicles and investigate the mechanisms controlling their biogenesis. Recent studies have confirmed the involvement of the lipid-modifying enzyme Phospholipase D2 (PLD2) in exosome generation and its downstream role in the GTPase ARF6. This review aims to recapitulate our current understanding of the role of PLD2 and its product, Phosphatidic Acid (PA), in exosome biogenesis and propose further research into the potential central roles of these molecules in the biology of these extracellular vesicles.

Phospholipase D and phosphatidic acid play roles in the formation of multivesicular bodies (MVBs) and their movement to the cell membrane during the process of exosome biogenesis within cells.

Case 2

Role of sphingolipids in the biogenesis andbiological activity of extracellular vesicles.

Journal: Lipid Res.
Published: 2018

Extracellular Vesicles (EVs) are vesicles released by both eukaryotic and prokaryotic cells. They possess unique physiological functions, such as processing cellular components and waste, and also play a pathophysiological role in inflammation and degenerative diseases. The shared molecular mechanisms underlying EV biogenesis are evident across different pathological cellular contexts in eukaryotes, and inhibiting EV biogenesis may offer avenues for therapeutic research due to their potential therapeutic significance. However, sphingolipids and related enzymes have received limited attention in the context of EV biogenesis and release. This review aims to outline the involvement of sphingolipids in extracellular vesicle biogenesis by shaping membrane curvature and through interactions with recipient cell membranes.

The review begins by describing how acidic and neutral sphingomyelinases promote the biogenesis of cytoplasmic vesicles in the cytoplasmic membrane and intracellular bodies, respectively, by generating sphingolipid ceramides. The involvement of other sphingolipids, such as ceramide phosphate and lactosylceramide, in extracellular vesicle formation and cargo sorting is then discussed. Finally, the review looks ahead to a series of biological effects mediated by changes in sphingolipid levels in recipient cells that may be modulated by alterations in receptor cell sphingolipid levels, suggesting a range of potential impacts on extracellular vesicle biology.

Over the past decade, progress has been made in elucidating the formation of EVs, and the fundamental role of ceramide in generating membrane curvature (away from the cytoplasmic direction) has been established, which is essential for both exosome and microvesicle biogenesis. The review discusses examples of EV biogenesis, where n- and a-SMase (neutral and acidic sphingomyelinase) have been extensively studied to mediate the release of exosomes and microvesicles, respectively. Specifically, the definitive role of a-SMase has been confirmed in MV release stimulated by various surface receptors or under stress conditions. However, the inhibition of n-SMase by GW4869 is a more complex situation, enhancing rather than blocking the production of microvesicles on the surface of epithelial cells.

Collective evidence suggests that sphingolipids (SLs) and their enzymes play a significant role in EVs, despite their lower content and uncertain species. Aside from serving as potential disease biomarkers in cancer and inflammatory diseases, vesicular SLs and their metabolic enzymes have been shown to promote EV function by influencing SL levels in recipient cells. Interestingly, EVs may also affect the activity of SLs in recipient cells by transferring miRNAs targeting SL metabolism, as confirmed by transcriptomic analysis of EVs produced by prokaryotic cells. We anticipate that characterizing EVs in different cellular physiologies in the future will contribute to understanding these highly conserved mammalian cell communication mediators, and exploring their biogenesis and signal transduction functions will have broad implications.