Summary of Study ST003259

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 PR002015. The data can be accessed directly via it's Project DOI: 10.21228/M85238 This work is supported by NIH grant, U2C- DK119886.

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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.

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Study IDST003259
Study TitleExploration of EMT-dependent changes of phosphatidylethanolamine profiles in KPC allografts
Study SummaryCryo-conserved tumors from subcutaneous allografts as described (Krebs et al. 2017, DOI: 10.1038/ncb3513) were retrieved. Three tumors derived from mesenchymal KPC cell lines (lines KPC550 and KPC701) and epithelial KPC cell lines (lines KPC438 and KPC661) were analyzed for their phosphatidylethanolamine profile by UPLC-MS/MS.
Institute
University of Innsbruck
DepartmentMichael Popp Institute
Last NameKoeberle
First NameAndreas
AddressMitterweg 24, Innsbruck, Tyrol, 6020, Austria
EmailAndreas.Koeberle@uibk.ac.at
Phone+43 512 507 57903
Submit Date2024-05-27
Raw Data AvailableYes
Raw Data File Type(s)wiff
Analysis Type DetailLC-MS
Release Date2024-07-05
Release Version1
Andreas Koeberle Andreas Koeberle
https://dx.doi.org/10.21228/M85238
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002015
Project DOI:doi: 10.21228/M85238
Project Title:Zeb1-mediated control of the phospholipid PUFA/MUFA ratio in EMT/plasticity-associated 1 cancer cell ferroptosis
Project Summary:Therapy resistance and metastasis, the most fatal steps in cancer, are often triggered by a (partial) activation of the epithelial-mesenchymal-transition (EMT)-program. A mesenchymal phenotype predisposes to ferroptosis, a cell death pathway exerted by an iron and oxygen-radical mediated peroxidation of phospholipids containing polyunsaturated fatty acids (PUFAs). We here describe that various forms of EMT-activation increase ferroptosis-susceptibility in cancer cells, which depends on the EMT-transcription factor Zeb1. To further investigate the underlying mechanisms of an EMT/Zeb1-coupled ferroptosis sensitivity, we analyzed key determinants of ferroptotic cell death, focusing on the proportion and (per)oxidation of fatty acid species in phospholipid subclasses. Using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), we demonstrate that GPX4 inhibition in human breast cancer MDA-MB-231 cells (Zeb1high) led to a rapid (per)oxidation of PUFA-containing phospholipids (oxPL), which is absent in cells depleted of Zeb1 (shZeb1). Mechanistically, Zeb1 increases the ratio of phospholipids containing pro-ferroptotic PUFAs over cyto-protective monounsaturated fatty acids (MUFAs) in MDA-MB-231 cells, tumor-derived pancreatic cancer KPC cells as well as mice tumor allografts via the modulation of crucial lipogenic enzymes.
Institute:University of Innsbruck
Department:Michael Popp Institute
Last Name:Koeberle
First Name:Andreas
Address:Mitterweg 24, Innsbruck, Tyrol, 6020, Austria
Email:Andreas.Koeberle@uibk.ac.at
Phone:+43 512 507 57903
Funding Source:the Austrian Science Fund (FWF) (P 36299), the German Research Council (GRK 1715), and the Phospholipid Research Center (Grant Number AKO‐2019‐070/2‐1, AKO-2O22-100/2-2), the Tyrolean Science Fund (TWF) (F.33467/7-2021).
Publications:in revision
Contributors:Zhigang Rao, Jie Zhang, André Gollowitzer, Leonhard Bereuter, Andreas Koeberle

Subject:

Subject ID:SU003379
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammals

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Phenotype
SA354070230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_01Pancreatic Cancer cell allografts epithelial/mixed
SA354071230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_02Pancreatic Cancer cell allografts epithelial/mixed
SA354072230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_03Pancreatic Cancer cell allografts epithelial/mixed
SA354073230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_04Pancreatic Cancer cell allografts mesenchymal
SA354074230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_05Pancreatic Cancer cell allografts mesenchymal
SA354075230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_06Pancreatic Cancer cell allografts mesenchymal
Showing results 1 to 6 of 6

Collection:

Collection ID:CO003372
Collection Summary:Cryo-conserved tumors from subcutaneous allografts described in (Krebs et al. 2017, DOI: 10.1038/ncb3513) were retrieved.
Sample Type:Tumor allograft
Storage Conditions:-80℃

Treatment:

Treatment ID:TR003388
Treatment Summary:KPC cells were subcutaneously injected into the flanks of C57BL/6 mice for engraftment as described (Krebs et al. 2017, DOI: 10.1038/ncb3513). Three tumor allografts derived from mesenchymal KPC cell lines (lines KPC550 and KPC701) and epithelial KPC cell lines (lines KPC438 and KPC661) were collected, cryo-conserved and stored at -80°C.

Sample Preparation:

Sampleprep ID:SP003386
Sampleprep Summary:Phospholipids were extracted from allograft tumor tissue by successive addition of PBS pH 7.4, methanol, chloroform, and saline to a final ratio of 14:34:35:17. Evaporation of the organic layer yielded a lipid film that was dissolved in methanol and subjected to UPLC-MS/MS.
Extract Storage:-80℃

Combined analysis:

Analysis ID AN005343
Analysis type MS
Chromatography type Reversed phase
Chromatography system Waters Acquity H-Class
Column Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex 6500+ QTrap
Ion Mode NEGATIVE
Units relative intensities

Chromatography:

Chromatography ID:CH004045
Chromatography Summary:Chromatographic separation of phospholipids was carried out on an Acquity BEH C8 column (1.7 μm, 2.1×100 mm, Waters, Milford, MA) using an Acquity UHPLC.
Instrument Name:Waters Acquity H-Class
Column Name:Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)
Column Temperature:45
Flow Gradient:The gradient was ramped from 75 to 85% B over 5 min and further increased to 100% B within 2 min, followed by isocratic elution for another 2 min.
Flow Rate:0.75 mL/min
Solvent A:90% Water, 10% Acetonitrile; 2 mM ammonium acetate
Solvent B:5% Water, 95% Acetonitrile; 2 mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS005073
Analysis ID:AN005343
Instrument Name:ABI Sciex 6500+ QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Targeted MRM with pre-optimized settings and subsequent automated integration of selected signals using Analyst 1.6.3 or Analyst 1.7.1 (Sciex). Phospholipids were analyzed in the negative ion mode, and both fatty acid anion fragments were detected by multiple reaction monitoring (MRM). For quantitation, the mean of both transitions was calculated. For the calculation of relative intensities (i.e., the proportion of lipids), all analyzed signals within the subgroup were summarized (= 100%), and the signals of individual lipid species or lipid subfractions are expressed as percentage of this sum.
Ion Mode:NEGATIVE
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