{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003257","ANALYSIS_ID":"AN005342","PROJECT_ID":"PR002015","VERSION":"1","CREATED_ON":"May 27, 2024, 10:51 am"},

"PROJECT":{"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"},

"STUDY":{"STUDY_TITLE":"Exploration of EMT-dependent changes in the lipidome of KPC cells","STUDY_SUMMARY":"KPC cell lines that are mesenchymal (lines KPC550 and KPC701), mesenchymal/epithelial mixed (lines KPC524 and KPC438), epithelial (lines KPC661 and KPC792) and KPC cell lines with Zeb1 knockout (lines KPCZ519, KPCZ436, KPCZ426) were analyzed for their phospholipid profile by UPLC-MS/MS.","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"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"KPC550",
"Sample ID":"200326_Brabletz_EMT_ZEB1_1_KPC550_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC550",
"Sample ID":"200326_Brabletz_EMT_ZEB1_2_KPC550_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC550",
"Sample ID":"200326_Brabletz_EMT_ZEB1_3_KPC550_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC701",
"Sample ID":"200326_Brabletz_EMT_ZEB1_4_KPC701_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC701",
"Sample ID":"200326_Brabletz_EMT_ZEB1_5_KPC701_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC701",
"Sample ID":"200326_Brabletz_EMT_ZEB1_6_KPC701_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC524",
"Sample ID":"200326_Brabletz_EMT_ZEB1_7_KPC524_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC524",
"Sample ID":"200326_Brabletz_EMT_ZEB1_8_KPC524_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC524",
"Sample ID":"200326_Brabletz_EMT_ZEB1_9_KPC524_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC438",
"Sample ID":"200326_Brabletz_EMT_ZEB1_10_KPC438_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC438",
"Sample ID":"200326_Brabletz_EMT_ZEB1_11_KPC438_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC438",
"Sample ID":"200326_Brabletz_EMT_ZEB1_12_KPC438_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"mesenchymal/epithelial mixed"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC661",
"Sample ID":"200326_Brabletz_EMT_ZEB1_13_KPC661_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC661",
"Sample ID":"200326_Brabletz_EMT_ZEB1_14_KPC661_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC661",
"Sample ID":"200326_Brabletz_EMT_ZEB1_15_KPC661_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC792",
"Sample ID":"200326_Brabletz_EMT_ZEB1_16_KPC792_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC792",
"Sample ID":"200326_Brabletz_EMT_ZEB1_17_KPC792_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPC792",
"Sample ID":"200326_Brabletz_EMT_ZEB1_18_KPC792_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"epithelial"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ519",
"Sample ID":"200326_Brabletz_EMT_ZEB1_19_KPCZ519_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ519",
"Sample ID":"200326_Brabletz_EMT_ZEB1_20_KPCZ519_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ519",
"Sample ID":"200326_Brabletz_EMT_ZEB1_21_KPCZ519_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ436",
"Sample ID":"200326_Brabletz_EMT_ZEB1_22_KPCZ346_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ436",
"Sample ID":"200326_Brabletz_EMT_ZEB1_23_KPCZ346_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ436",
"Sample ID":"200326_Brabletz_EMT_ZEB1_24_KPCZ346_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ426",
"Sample ID":"200326_Brabletz_EMT_ZEB1_25_KPCZ426_1",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ426",
"Sample ID":"200326_Brabletz_EMT_ZEB1_26_KPCZ426_2",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
},
{
"Subject ID":"KPCZ426",
"Sample ID":"200326_Brabletz_EMT_ZEB1_27_KPCZ426_3",
"Factors":{"Sample source":"KPC cells","Phenotype":"Zeb1 knockout"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name for PE)":"200326_Brabletz_EMT_ZEB1_PE_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PC)":"200326_Brabletz_EMT_ZEB1_PC_dil_1_10.wiff","RAW_FILE_NAME(Raw file name for PI)":"200326_Brabletz_EMT_ZEB1_PI_dil_1_10.wiff"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"• Cultured cells were washed, trypsinized, counted and flash-frozen in liquid N2 and stored at -80°C.","SAMPLE_TYPE":"Tumor cells","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Tumor-derived pancreatic cancer KPC cells that are mesenchymal (lines KPC550 and KPC701), mesenchymal/epithelial mixed (lines KPC524 and KPC438), epithelial (lines KPC661 and KPC792) and KPC cell lines with Zeb1 knockout (lines KPCZ519, KPCZ436, KPCZ426) were obtained from tumors of KPC mice as described (Krebs et al. 2017, DOI: 10.1038/ncb3513)."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Phospholipids were extracted from cell pellets 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℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity H-Class","COLUMN_NAME":"Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)","SOLVENT_A":"water/acetonitrile 90/10, 2 mM ammonium acetate","SOLVENT_B":"water/acetonitrile 5/95, 2 mM ammonium acetate","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","COLUMN_TEMPERATURE":"45"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"ABI Sciex 6500+","INSTRUMENT_TYPE":"QTRAP","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","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)."},

"MS_METABOLITE_DATA":{
"Units":"relative intensities",

"Data":[{"Metabolite":"PE 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}

}