Summary of Study ST003504
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002150. The data can be accessed directly via it's Project DOI: 10.21228/M8Q82X This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST003504 |
| Study Title | Methionine-SAM metabolism-dependent ubiquinone synthesis is crucial for ROS accumulation in ferroptosis induction |
| Study Summary | Ferroptosis is a cell death modality in which iron-dependent lipid peroxides accumulate on cell membranes. Cysteine, a limiting substrate for the glutathione system that neutralizes lipid peroxidation and prevents ferroptosis, can be converted by cystine reduction or synthesized from methionine. However, accumulating evidence shows methionine-based cysteine synthesis fails to effectively rescue intracellular cysteine levels upon cystine deprivation and is unable to inhibit ferroptosis. Here, we report that methionine-based cysteine synthesis is tissue-specific. Unexpectedly, we find that rather than inhibiting ferroptosis, methionine in fact plays an essential role during cystine deprivation-induced ferroptosis. Methionine-derived S-adenosylmethionine (SAM) contributes to methylation-dependent ubiquinone synthesis, which leads to lipid peroxides accumulation and subsequent ferroptosis. Moreover, SAM supplementation synergizes with imidazole ketone erastin in a tumor growth suppression mouse model. Inhibiting the enzyme that converts methionine to SAM protects heart tissue from doxorubicin-induced and ferroptosis-driven cardiomyopathy. This study broadens our understanding about the intersection of amino acid metabolism and ferroptosis regulation, providing insight into the underlying mechanisms and suggesting the methionine-SAM axis is a promising therapeutic strategy to treat ferroptosis-related diseases. To test, we firstly performed a metabolomics analysis of HT1080 cells that were cultured in medium lacking cystine or lacking both cystine and methionine. The metabolomics results containing full profile of the metabolites was subjected to cluster assay, and the triplicates of each group were automatically clustered together, indicating better homogeneity of the data in each condition. Then, we filtrated the metabolomics data with the threshold fold change (FC) ≥2 and P≤ 0.05. Only the RP Pos method results are used/reported in this study. The results showed that the cellular cysteine and GSH levels were expectedly depressed upon cystine limitation; meanwhile, once methionine was restricted, cellular SAM level was significantly reduced. This highlights the decisive regulatory role of methionine on the metabolite levels of SAM. |
| Institute | Northeast Normal University |
| Last Name | xia |
| First Name | chaoyi |
| Address | 5268 Renmin Street, changchun, ji lin, 130024, China |
| xiacy834@nenu.edu.cn | |
| Phone | 18166850252 |
| Submit Date | 2024-09-26 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-09-30 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002150 |
| Project DOI: | doi: 10.21228/M8Q82X |
| Project Title: | Methionine-SAM metabolism-dependent ubiquinone synthesis is crucial for ROS accumulation in ferroptosis induction |
| Project Summary: | Ferroptosis is a cell death modality in which iron-dependent lipid peroxides accumulate on cell membranes. Cysteine, a limiting substrate for the glutathione system that neutralizes lipid peroxidation and prevents ferroptosis, can be converted by cystine reduction or synthesized from methionine. However, accumulating evidence shows methionine-based cysteine synthesis fails to effectively rescue intracellular cysteine levels upon cystine deprivation and is unable to inhibit ferroptosis. Here, we report that methionine-based cysteine synthesis is tissue-specific. Unexpectedly, we find that rather than inhibiting ferroptosis, methionine in fact plays an essential role during cystine deprivation-induced ferroptosis. Methionine-derived S-adenosylmethionine (SAM) contributes to methylation-dependent ubiquinone synthesis, which leads to lipid peroxides accumulation and subsequent ferroptosis. Moreover, SAM supplementation synergizes with imidazole ketone erastin in a tumor growth suppression mouse model. Inhibiting the enzyme that converts methionine to SAM protects heart tissue from doxorubicin-induced and ferroptosis-driven cardiomyopathy. This study broadens our understanding about the intersection of amino acid metabolism and ferroptosis regulation, providing insight into the underlying mechanisms and suggesting the methionine-SAM axis is a promising therapeutic strategy to treat ferroptosis-related diseases. |
| Institute: | Northeast Normal University |
| Last Name: | xia |
| First Name: | chaoyi |
| Address: | 5268 Renmin Street, changchun, ji lin, 130024, China |
| Email: | xiacy834@nenu.edu.cn |
| Phone: | 18166850252 |
Subject:
| Subject ID: | SU003633 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Male |
Factors:
Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Treatment |
|---|---|---|---|
| SA385788 | XiaChaoYi_A4_RP pos | HT1080 human epithelial cancer cells | +CC+Met |
| SA385789 | XiaChaoYi_A7_RP pos | HT1080 human epithelial cancer cells | +CC+Met |
| SA385790 | XiaChaoYi_A10_RP pos | HT1080 human epithelial cancer cells | +CC+Met |
| SA385791 | XiaChaoYi_A5_RP pos | HT1080 human epithelial cancer cells | -CC+Met |
| SA385792 | XiaChaoYi_A8_RP pos | HT1080 human epithelial cancer cells | -CC+Met |
| SA385793 | XiaChaoYi_A11_RP pos | HT1080 human epithelial cancer cells | -CC+Met |
| SA385794 | XiaChaoYi_A6_RP pos | HT1080 human epithelial cancer cells | -CC-Met |
| SA385795 | XiaChaoYi_A9_RP pos | HT1080 human epithelial cancer cells | -CC-Met |
| SA385796 | XiaChaoYi_A12_RP pos | HT1080 human epithelial cancer cells | -CC-Met |
| Showing results 1 to 9 of 9 |
Collection:
| Collection ID: | CO003626 |
| Collection Summary: | Cells were seeded at a density of 2×106 per well in a 10 cm glass bottom cell culture dish and grown for 12 h in DMEM supplemented with 10% Fetal Bovine Serum（FBS）, 1% Penicillin-Streptomycin(P/S). |
| Collection Protocol Filename: | 2024-NC.pdf |
| Sample Type: | HT1080 Human epithelial cancer cells |
Treatment:
| Treatment ID: | TR003642 |
| Treatment Summary: | HT1080 cells were cultured in the medium ± cystine or ± methionine as indicated for 10 h. The metabolites levels were measured by UHPLC-HRMS. |
Sample Preparation:
| Sampleprep ID: | SP003640 |
| Sampleprep Summary: | Cells were trypsinized, washed and resuspended in methanol: acetonitrile: ddH2O2 (2:2:1, v/v) after indicated treatment. Then subjected to LC/MS analysis at LipidALL Technologies (Changzhou, China). Briefly, polar metabolites were extracted using 1000 µl of ice-cold methanol: H2O (4:1, v/v), and incubated at 2000 × g for 30 min at 4oC. At the end of the incubation, samples were centrifuged for 10 min at 13,000 × g at 4oC. Clean supernatant was transferred to a new tube. Extracts were dried in a centrifugal concentrator. |
| Sampleprep Protocol Filename: | 2024-NC.pdf |
Combined analysis:
| Analysis ID | AN005752 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II |
| Column | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Triple TOF |
| MS instrument name | ABI Sciex 5600+ TripleTOF |
| Ion Mode | POSITIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH004366 |
| Methods Filename: | 2024-NC.pdf |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0 - 1.0 min with 2% B, 1.0 - 6.0 min with 2% - 42% B, 6.0 - 8.0 min with 42% - 65% B, 8.0 - 10.0 min with 65% - 76% B, 10.0 - 11.0 min with 76% - 100% B, 11.0 - 14.0 min with 100% - 100% B. |
| Flow Rate: | 0.35 mL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005475 |
| Analysis ID: | AN005752 |
| Instrument Name: | ABI Sciex 5600+ TripleTOF |
| Instrument Type: | Triple TOF |
| MS Type: | ESI |
| MS Comments: | MS parameters for detection were: ESI source voltage positive ion mode 5.5 kV, negative ion mode -4.5 kV; vaporizer temperature, 450°C; drying gas (N2) pressure, 50 psi; nebulizer gas (N2) pressure, 50 psi; curtain gas (N2) pressure, 35 psi; The scan range was m/z 60-800. Information-dependent acquisition mode was used for MS/MS analyses of the metabolites. Collision energy was set at (±) 35 ± 15 eV. Data acquisition and processing were performed using Analyst® TF 1.7.1 Software (AB Sciex, Concord, ON, Canada). |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | 2024-NC.pdf |
