Summary of Study ST002115
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 PR001340. The data can be accessed directly via it's Project DOI: 10.21228/M8HD8Q 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 | ST002115 |
| Study Title | LC-MS analysis of metabolic changes induced by GPX4 inhibitor treatment in cultured HT1080 cells |
| Study Summary | HT1080 cells were treated with vehicle (DMSO), RSL3 (10 micromolar), ML210 (10 micromolar), or ML162 (10 micromolar) for 2 hours. Cellular metabolites were then extracted and analyzed by LC-MS. |
| Institute | University of Texas MD Anderson Cancer Center |
| Last Name | Gan |
| First Name | Boyi |
| Address | 6565 MD Anderson Blvd, Houston TX, 77030 |
| bgan@mdanderson.org | |
| Phone | 713-792-8653 |
| Submit Date | 2022-03-02 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML |
| Analysis Type Detail | LC-MS |
| Release Date | 2022-04-14 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001340 |
| Project DOI: | doi: 10.21228/M8HD8Q |
| Project Title: | A ferroptosis defense mechanism mediated by glycerol 3-phosphate dehydrogenase 2 in mitochondria |
| Project Summary: | Mechanisms of defense against ferroptosis (an iron-dependent form of cell death induced by lipid peroxidation) in cellular organelles remain poorly understood, hindering our ability to target ferroptosis in disease treatment. In this study, metabolomic analyses revealed that treatment of cancer cells with glutathione peroxidase 4 (GPX4) inhibitors results in intracellular glycerol 3-phosphate (G3P) depletion. We further showed that supplementation of cancer cells with G3P attenuates ferroptosis induced by GPX4 inhibitors in a G3P dehydrogenase 2 (GPD2)-dependent manner; GPD2 deletion sensitizes cancer cells to GPX4 inhibition-induced mitochondrial lipid peroxidation and ferroptosis, and combined deletion of GPX4 and GPD2 synergistically suppresses tumor growth by inducing ferroptosis in vivo. Mechanistically, inner mitochondrial membrane-localized GPD2 couples G3P oxidation with ubiquinone reduction to ubiquinol, which acts as a radical-trapping antioxidant to suppress ferroptosis in mitochondria. Taken together, these results reveal that GPD2 participates in ferroptosis defense in mitochondria by generating ubiquinol. |
| Institute: | University of Texas MD Anderson Cancer Center |
| Last Name: | Gan |
| First Name: | Boyi |
| Address: | 6565 MD Anderson Blvd, Houston, TX 77030 |
| Email: | bgan@mdanderson.org |
| Phone: | 713-792-8653 |
Subject:
| Subject ID: | SU002231 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HT1080 |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA205788 | HT1080_DMSO_01 | DMSO |
| SA205789 | HT1080_DMSO_03 | DMSO |
| SA205790 | HT1080_DMSO_02 | DMSO |
| SA205791 | HT1080_ML162_03 | ML162 |
| SA205792 | HT1080_ML162_01 | ML162 |
| SA205793 | HT1080_ML162_02 | ML162 |
| SA205794 | HT1080_ML210_03 | ML210 |
| SA205795 | HT1080_ML210_01 | ML210 |
| SA205796 | HT1080_ML210_02 | ML210 |
| SA205797 | HT1080_RSL3_03 | RSL3 |
| SA205798 | HT1080_RSL3_02 | RSL3 |
| SA205799 | HT1080_RSL3_01 | RSL3 |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO002224 |
| Collection Summary: | Metabolites were extracted from cells in 35 mm culture plates by rapidly aspirating the culture medium and incubating the plates with 0.6 ml of an 80% methanol: 20% water mixture on a cold block on dry ice for 15 min. Next, the cell material was scraped into Eppendorf tubes pre-chilled on ice. After centrifugation at 13,000 RCF for 5 min at 4 °C, the supernatant was collected into a fresh tube and stored on dry ice until analysis. |
| Sample Type: | Cultured cells |
| Storage Conditions: | -80? |
Treatment:
| Treatment ID: | TR002243 |
| Treatment Summary: | Cells were seeded in 35-mm culture plates. When the cell confluence reached 70-80%, cells were treated with RSL3, ML210, or ML162 for 2 hours. |
Sample Preparation:
| Sampleprep ID: | SP002237 |
| Sampleprep Summary: | For analysis by reverse phase chromatography, just before analysis, 500 µL of extract was dried under a nitrogen gas flow and then resuspended in 100 µL of water. For analysis by HILIC chromatography, the extracts were analyzed directly. |
| Processing Storage Conditions: | 4? |
| Extract Storage: | 4? |
Combined analysis:
| Analysis ID | AN003513 | AN003514 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | HILIC |
| Chromatography system | Thermo Accela 1250 | Thermo Accela 1250 |
| Column | Phenomenex Synergi Hydro RP 100 A (100 x 2mm,2.5um) | Waters XBridge BEH Amide (150 x 2.1mm,2.5um,100A) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Exactive Orbitrap | Thermo Exactive Orbitrap |
| Ion Mode | NEGATIVE | NEGATIVE |
| Units | Peak area (top) | Peak area (top) |
Chromatography:
| Chromatography ID: | CH002593 |
| Chromatography Summary: | The gradient was 0 min, 0% B; 2.5 min, 0% B; 5 min, 20% B; 7.5 min, 20% B; 13 min, 55% B; 15.5 min, 95% B; 18.5 min, 95% B; 19 min, 0% B; and 25 min, 0% B. Solvent A was 10 mM tributylamine and 15 mm acetic acid in water; Solvent B was methanol. The injection volume was 10 µL. |
| Instrument Name: | Thermo Accela 1250 |
| Column Name: | Phenomenex Synergi Hydro RP 100 A (100 x 2mm,2.5um) |
| Column Temperature: | 40 |
| Flow Rate: | 200 |
| Solvent A: | 100% water; 15 mM acetic acid; 10 mM tributylamine |
| Solvent B: | 100% methanol |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH002594 |
| Chromatography Summary: | The gradient was 0 min, 85% B; 2 min, 85% B; 3 min, 80% B; 5 min, 80% B; 6 min, 75% B; 7 min, 75% B; 8 min, 70% B; 9 min, 70% B; 10 min, 50% B; 12 min, 50% B; 13 min, 25% B; 16 min, 25% B; 18 min, 0% B; 23 min, 0% B; 24 min, 85% B; 30 min, 85% B. The injection volume was 5 µL. |
| Instrument Name: | Thermo Accela 1250 |
| Column Name: | Waters XBridge BEH Amide (150 x 2.1mm,2.5um,100A) |
| Column Temperature: | 40 |
| Flow Rate: | 150 |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate, pH 9.4 |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003271 |
| Analysis ID: | AN003513 |
| Instrument Name: | Thermo Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The scan range was 80-1000 m/z. Raw data files were converted to mzXML format using msconvert (ProteoWizard). Data was analyzed in the MAVEN software suite and metabolite assignments were made using a previously generated list of retention times derived from pure standard solutions. |
| Ion Mode: | NEGATIVE |
| MS ID: | MS003272 |
| Analysis ID: | AN003514 |
| Instrument Name: | Thermo Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The scan range was 80-1000 m/z. Raw data files were converted to mzXML format using msconvert (ProteoWizard). Data was analyzed in the MAVEN software suite and metabolite assignments were made using a previously generated list of retention times derived from pure standard solutions. |
| Ion Mode: | NEGATIVE |
