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What Is Targeted Lipidomics
Lipidomics is a discipline that analyzes and identifies lipid metabolites and their interacting molecules in organisms, tissues or cells to understand the structure and function of lipids, and then reveals the relationship between lipid metabolism and the physiological and pathological processes of cells, organs and even organisms. With the advancement of mass spectrometry, lipidomics has shown a wide range of applications in the identification of lipid biomarkers, diagnosis of diseases, discovery of drug targets and lead compounds, and study of drug mechanisms of action.
Targeted lipidomics is based on high-resolution mass spectrometry and isotope internal standard, and uses PRM (parallel reaction monitoring) targeted analysis technology to specifically acquire the signals of multiple lipid molecules (such as dozens of target lipid molecules) simultaneously to obtain their absolute contents and meet the needs of targeted detection and validation of target lipids.
Lipids are hydrophobic or amphipathic small molecules which include fats, waxes, sterols, fat-soluble vitamins (such as vitamins A, D, E and K), monoglycerides, diglycerides and phospholipids. The crucial role of lipids in biological physiology is evident not only in energy storage and structural components of cellular membranes, but also in signal transduction, membrane trafficking and morphogenesis. It is known that mammalian cells contain 1000-2000 types of lipids, but there are many more than that, and it is conservatively estimated that the major lipid species theoretically contain nearly 200000 lipids. Lipids can be broadly classified into three categories, namely, polar lipids including glycerophospholipids and sphingolipids, and nonpolar lipids including glycerol esters and other lipid metabolites such as fatty acids, which are the precursors of biologically active lipid molecules.
The National Institutes of Health (NIH) classifies lipids into eight specific categories including fattyacids, sphingolipids, glycerolipids, glycerophosph1ipids, sterol lipids, saecharolipids, pregnenolipids, and polyketides.
Lipidomics Analysis Workflow
Equipped with advanced tandem mass spectrometry for the determination of lipid composition, powerful bioinformatic analytic systems, and a panel of experienced scientists and technicians, Creative Proteomics provide customer-tailored service with rapid analysis procedures and easy to read report, to speed up your scientific research.
Technology Platform
With an integrated set of separation, characterization, identification and quantification system, featured with excellent robustness & reproducibility, high and ultra-sensitivity, Creative Proteomics provides reliable, rapid and cost-effective targeted lipidomics services as below.
Featured Lipid Analysis Service
Fatty Acids Analysis Service
Fatty Acids Derivatives Analysis Service
Fatty Acids Metabolism Analysis Service
Glycerolipids Analysis Service
Phospholipidomics Service
Glycerophospholipids Analysis Service
Sphingolipids
Isoprenoids
Sterols
Other Analysis Services
Data Analysis
| Standard | Analysis content | |
|---|---|---|
| Standard analysis | Quality control | Get high quality data |
| Method evaluation | Assess the quality of the established method | |
| Difference analysis (histogram, violin plot, etc.) | Assess group differences for each lipid molecules | |
| Advanced analysis | Cluster Analysis | Exploring the content trend pattern of lipid molecules |
| Machine learning | Biomarkers screening with good diagnostic performance | |
Technical Advantages
- Platform advantages: orbitrap mass analyzer, ultra-high resolution mass spectrometry, high-quality data
- Wide applicability: no species restriction, no standard product restriction
- High throughput: detect dozens of lipid molecules at one time, saving samples and costs
- Strong quantitative ability: the sensitivity can reach ppm level, and the linear range can reach 5-6 orders of magnitude
- State of art facilities
- Constantly optimized protocol and analytical software
- Professional experiment design
- Quick turnaround time
Sample requirement
| Sample Type | Minimum Sample Amount |
|---|---|
| Cells | Not less than 1x10^7 cells |
| Tissues | Not less than 250 mg |
| Serum, Plasma | Not less than 300 μL |
| Feces and Intestinal Contents | Not less than 200 mg |
| Microorganisms (Bacteria, Fungi, etc.) | Not less than 500 mg, not less than 10^7 CFU/mL |
| Other Liquid Samples | "Urine not less than 1 mL, Saliva not less than 500 μL, |
| Amniotic Fluid, Bile, Cerebrospinal Fluid, Lymph Fluid, etc., not less than 300 μL" |
Ordering Procedure
Retinol Saturase Mediates Retinoid Metabolism to Impair a Ferroptosis Defense System in Cancer Cells
Journal: Cancer Res
Published: 2023
Research Methods: Targeted Lipidomics, qRT-PCR, LC-MS, Immunohistochemistry
Background
Ferroptosis is an iron-dependent regulated cell death pathway caused by lipid peroxidation overload on the cell membrane. In recent years, ferroptosis has garnered attention and has been extensively studied. This research employs methods such as targeted lipidomics and immunohistochemistry to explore the induction of ferroptosis in tumor cells, suggesting its potential as a therapeutic approach. However, the multifaceted regulation through various pathways from different dimensions still requires further exploration. The concept of ferroptosis and its underlying mechanisms provide a new direction for advancing targeted therapies in cancer treatment.
Results
Targeted lipidomics reveals that RETSAT depletion promotes monounsaturated fatty acid generation Targeted lipidomics discovered a significant increase in fatty acids and phospholipids in A549 cells carrying RETSAT-KO. The lipidomic results align with the increased resistance of RETSAT-KO cell lines to iron-deficiency anemia, indicating that the RETSAT-RAR pathway mediates ferroptosis by promoting the generation of monounsaturated fatty acids (or acyls) (MUFA)
Figure: RETSAT-RAR Pathway Promotes Ferroptosis Anemia through SCD-Mediated MUFA Generation
The RETSAT-RAR pathway promotes sensitivity to ferroptosis anemia by SCD-mediated MUFA generation
Targeted lipidomics revealed that RETSAT-KO and RAR activation induced the expression of SCD and the production of MUFA. RARα binding sites (RARE), RETSAT depletion, and RAR agonist activation led to transcriptional activation, confirming that RAR directly activates SCD transcription by binding to its promoter. Inhibition of RETSAT suppresses the activity of the RAR signaling pathway, slowing down RAR-targeted transcription of SCD, consequently resulting in reduced MUFA generation.
Conclusions
The study revealed the regulatory role of retinoids in ferroptosis is associated with lipid metabolism. Retinoids possess antioxidant capabilities that prevent cell membrane rupture caused by lipid peroxidation, ultimately terminating ferroptosis. Through investigations like targeted lipidomics, it was also found that retinoids can promote the generation of monounsaturated fatty acids. Moreover, intracellular retinoid metabolism can alter tumor iron descent and is correlated with the prognosis of LUAD patients.
This study, employing targeted metabolomics, immunohistochemistry, and other methodologies, unveiled the antagonistic mechanism of retinoids against tumor cell ferroptosis. This discovery contributes to the exploration of therapeutic strategies that target tumor cell retinoid metabolism to induce ferroptosis, offering novel avenues and approaches for advancing targeted cancer therapies.
Reference
- Bi G, Liang J, Shan G, Bian Y, Chen Z, Huang Y, Lu T, Li M, Besskaya V, Zhao M, Fan H, Wang Q, Gan B, Zhan C. Retinol Saturase Mediates Retinoid Metabolism to Impair a Ferroptosis Defense System in Cancer Cells. Cancer Res. 2023 Jul 14;83(14):2387-2404.
