Summary of study ST001451

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 PR000997. The data can be accessed directly via it's Project DOI: 10.21228/M8TH75 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001451
Study TitleEleostearic acid effects on TAGs and oxLipids
Study SummaryQuantification of lipid species from cultured human MDA-MB-231 and BT549 cells with or without siRNA knockdown of ACSL1 and treated or not with eleostearic acid
Institute
Fox Chase Cancer Center
Last NamePeterson
First NameJeffrey
Address333 Cottman Avenue Philadelphia, PA 19111
EmailJeffrey.peterson@fccc.edu
Phone215-728-3568
Submit Date2020-08-20
Raw Data AvailableYes
Raw Data File Type(s).raw
Analysis Type DetailLC-MS
Release Date2020-09-10
Release Version1
Jeffrey Peterson Jeffrey Peterson
https://dx.doi.org/10.21228/M8TH75
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000997
Project DOI:doi: 10.21228/M8TH75
Project Title:ACSL1 role in conjugated PUFA metabolism
Project Type:MS quantitative lipidomic study
Project Summary:Ferroptosis is associated with lipid hydroperoxides generated by oxidation of polyunsaturated acyl chains. Lipid hydroperoxides are reduced by glutathione peroxidase 4 (GPX4) and GPX4 inhibitors induce ferroptosis. However, the therapeutic potential of triggering ferroptosis in cancer cells with polyunsaturated fatty acids is unknown. We identified conjugated linoleates including α-eleostearate (αESA) as novel ferroptosis inducers. αESA did not alter GPX4 activity but was incorporated into cellular lipids and promoted lipid peroxidation and cell death in diverse cancer cell types. αESA-triggered death was mediated by acyl-CoA synthetase long-chain isoform 1, which promoted αESA incorporation into neutral lipids including triacylglycerols. Interfering with triacylglycerol biosynthesis suppressed ferroptosis triggered by αESA.
Institute:Fox Chase Cancer Center
Last Name:Peterson
First Name:Jeffrey
Address:333 Cottman Avenue, Philadelphia, PA, 19111, USA
Email:Jeffrey.peterson@fccc.edu
Phone:215-728-3568
Funding Source:DOD, NIH
Publications:in process
Contributors:Valarian Kagan

Subject:

Subject ID:SU001525
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Gender:Female

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id siRNA Treatment (8hrs)
SA124011Nov5_2018_036ACSL1 eleostearic acid (25µM)
SA124012Nov5_2018_037ACSL1 eleostearic acid (25µM)
SA124013Jun17_2020_071ACSL1 eleostearic acid (25µM)
SA124014Jun17_2020_073ACSL1 eleostearic acid (25µM)
SA124015Nov5_2018_035ACSL1 eleostearic acid (25µM)
SA124016Jun17_2020_069ACSL1 eleostearic acid (25µM)
SA124017Nov5_2018_033ACSL1 eleostearic acid (25µM)
SA124018Jun17_2020_063ACSL1 eleostearic acid (25µM)
SA124019Nov5_2018_034ACSL1 eleostearic acid (25µM)
SA124020Jun17_2020_065ACSL1 eleostearic acid (25µM)
SA124021Jun17_2020_067ACSL1 eleostearic acid (25µM)
SA124022Nov5_2018_032ACSL1 eleostearic acid (25µM)
SA123999Nov5_2018_029ACSL1 Vehicle (methanol)
SA124000Nov5_2018_027ACSL1 Vehicle (methanol)
SA124001Nov5_2018_026ACSL1 Vehicle (methanol)
SA124002Nov5_2018_030ACSL1 Vehicle (methanol)
SA124003Jun17_2020_055ACSL1 Vehicle (methanol)
SA124004Jun17_2020_051ACSL1 Vehicle (methanol)
SA124005Jun17_2020_053ACSL1 Vehicle (methanol)
SA124006Jun17_2020_057ACSL1 Vehicle (methanol)
SA124007Nov5_2018_031ACSL1 Vehicle (methanol)
SA124008Nov5_2018_028ACSL1 Vehicle (methanol)
SA124009Jun17_2020_059ACSL1 Vehicle (methanol)
SA124010Jun17_2020_061ACSL1 Vehicle (methanol)
SA124035Jun17_2020_045Control eleostearic acid (25µM)
SA124036Jun17_2020_043Control eleostearic acid (25µM)
SA124037Jun17_2020_041Control eleostearic acid (25µM)
SA124038Jun17_2020_039Control eleostearic acid (25µM)
SA124039Jun17_2020_047Control eleostearic acid (25µM)
SA124040Nov5_2018_022Control eleostearic acid (25µM)
SA124041Jun17_2020_049Control eleostearic acid (25µM)
SA124042Nov5_2018_025Control eleostearic acid (25µM)
SA124043Nov5_2018_024Control eleostearic acid (25µM)
SA124044Nov5_2018_023Control eleostearic acid (25µM)
SA124045Nov5_2018_021Control eleostearic acid (25µM)
SA124023Jun17_2020_037Control Vehicle (methanol)
SA124024Jun17_2020_035Control Vehicle (methanol)
SA124025Jun17_2020_027Control Vehicle (methanol)
SA124026Nov5_2018_020Control Vehicle (methanol)
SA124027Jun17_2020_033Control Vehicle (methanol)
SA124028Nov5_2018_019Control Vehicle (methanol)
SA124029Nov5_2018_018Control Vehicle (methanol)
SA124030Nov5_2018_016Control Vehicle (methanol)
SA124031Nov5_2018_017Control Vehicle (methanol)
SA124032Jun17_2020_029Control Vehicle (methanol)
SA124033Nov5_2018_015Control Vehicle (methanol)
SA124034Jun17_2020_031Control Vehicle (methanol)
Showing results 1 to 47 of 47

Collection:

Collection ID:CO001520
Collection Summary:BT-549 cells were transfected with ACSL1 or non-targeting siRNA 72 hours prior to incubation with 25 uM alpha eleostearic acid or vehicle (methanol) for 8 hours. After 8 hours, cells (~4-6 x 106 per replicate) were trypsinized, collected, washed once with phosphate-buffered saline, and frozen in liquid nitrogen.
Sample Type:Breast cancer cells
Storage Conditions:-80℃

Treatment:

Treatment ID:TR001540
Treatment Summary:Cells were treated in complete culture medium with 25uM alpha eleostearic acid or methanol (vehicle control) for eight hours
Treatment Compound:alpha eleostearic acid

Sample Preparation:

Sampleprep ID:SP001533
Sampleprep Summary:Cells were re-suspended in 25 mM HEPES buffer (pH 7.4) containing 200 μM DTPA and sonicated.

Combined analysis:

Analysis ID AN002425 AN002426
Analysis type MS MS
Chromatography type Normal phase Reversed phase
Chromatography system Thermo Dionex Ultimate 3000 Thermo Dionex Ultimate 3000
Column Phenomenex Luna Silica (150 x 2.0mm , 3um) Phenomenex Luna Silica (150 x 1.0mm , 3um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Fusion Orbitrap Thermo Q Exactive HF hybrid Orbitrap
Ion Mode NEGATIVE POSITIVE
Units pmol/mg pmol/umol

Chromatography:

Chromatography ID:CH001782
Chromatography Summary:Phospholipid protocol
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:Phenomenex Luna Silica (150 x 2.0mm , 3um)
Chromatography Type:Normal phase
  
Chromatography ID:CH001783
Chromatography Summary:Triacylglycerols
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:Phenomenex Luna Silica (150 x 1.0mm , 3um)
Chromatography Type:Reversed phase

MS:

MS ID:MS002262
Analysis ID:AN002425
Instrument Name:Thermo Fusion Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:Analysis of LC/MS data was performed using software package Compound Discoverer (ThermoFisher Scientific) with an in-house generated analysis workflow and oxidized phospholipid database.
Ion Mode:NEGATIVE
  
MS ID:MS002263
Analysis ID:AN002426
Instrument Name:Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:TAG cations were formed through molecular ammonium adduction (+NH4). Analysis was performed at a resolution of 140,000 for the full MS scan and 17,500 for the MS2 scan in a data-dependent mode. The scan range for MS analysis was 300–1200 m/z with a maximum injection time of 128 ms using one microscan. A maximum injection time of 500 ms was used for MS2 (high energy collisional dissociation (HCD)) analysis with collision energy set to 24. An isolation window of 1.0 Da was set for the MS and MS2 scans. Capillary spray voltage was set at 4.5 kV, and capillary temperature was 320 °C. Sheath gas was set to eight arbitrary units and the S-lens Rf level was set to 60. Standards for TAGs and their oxygenated metabolites were from Avanti Polar Lipids and Cayman Chemicals.
Ion Mode:POSITIVE
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