Summary of Study ST003261

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 PR002023. The data can be accessed directly via it's Project DOI: 10.21228/M8423Z 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	ST003261
Study Title	Exploration of RSL3 or Chlorido[N,N’-disalicylidene-1,2-phenylenediamine]iron(III) complex-induced changes in the phospholipid oxidation of MDA-MB-231 breast cancer cells
Study Summary	Chlorido[N,N’-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) exhibit potent anti-cancer properties through incompletely understood molecular mechanisms. Here, we treated human MDA-MB-231 triple-negative breast cancer cells with the glutathione peroxidase (GPX)4 inhibitor RSL3 or chlorido[N,N’-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs) and analyzed their oxidized phospholipid profile by targeted lipidomics. SCs induce extensive (hydroper)oxidation of arachidonic acid and adrenic acid in membrane phospholipids, particularly phosphatidylethanolamines (PE) and phosphatidylinositols (PC). In this process, SCs have demonstrated superior efficacy compared to the GPX4 inhibitor RSL3, an established ferroptosis inducer. Please note that one sample set was measured three times with the same sample-ID, but with different methods (oxPE, oxPC, oxPI), therefore each sub-class has their own raw-data file marked by their corresponding abbreviation (oxPE, oxPC, oxPI; e.g. "210309_MDA_Timecourse_RSL3_oxPE_dil_UD_Std_1ul_SFT.wiff", "210324_Rescue_Gust_compounds_oxPE_dil_UD_std_1ul_SFT.wiff", "210309_MDA_Timecourse_RSL3_oxPC_dil_UD_Std_1ul_SFT.wiff", "210324_Rescue_Gust_compounds_oxPC_dil_UD_std_1ul_SFT.wiff" or "210324_Rescue_Gust_compounds_oxPI_dil_UD_std_1ul_SFT.wiff", ).
Institute	University of Innsbruck
Last Name	Koeberle
First Name	Andreas
Address	Mitterweg 24, Innsbruck, Tyrol, 6020, Austria
Email	Andreas.Koeberle@uibk.ac.at
Phone	+43 512 507 57903
Submit Date	2024-06-12
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2024-06-27
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002023
Project DOI:	doi: 10.21228/M8423Z
Project Title:	Iron(III)-salophene catalyzes redox cycles that induce phospholipid peroxidation and deplete cancer cells of ferroptosis-protecting cofactors
Project Summary:	Ferroptosis, regulated by glutathione peroxidase 4 and redox cycles, offers new cancer treatment strategies. Chlorido[N,N’-disalicylidene-1,2-phenylenediamine]iron(III) complexes (SCs, compounds 1-3) exhibit potent anti-cancer effects by inducing ferroptosis, apoptosis, or necroptosis, including in therapy-resistant cancers. Our study shows that SCs favor ferroptosis in triple-negative breast cancer cells and are effective against invasive, chemo- or radioresistant cell lines. Redox lipidomics indicates that SCs initiate cell death through extensive oxidation of arachidonic and adrenic acids in membrane phospholipids. Mechanistically, SCs catalyze one-electron transfer reactions, reducing Fe(III) to Fe(II), forming oxo-bridged dimers, and generating organic radicals using hydrogen peroxide. This process depletes NADPH, oxidizes membrane phospholipids, and disrupts cellular detoxification of phospholipid hydroperoxides.
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:	Austrian Science Fund (FWF) (P 36299), Phospholipid Research Center Heidelberg (AKO-2022-100/2-2)
Publications:	DOI : https://doi.org/10.1016/j.redox.2024.103257
Contributors:	Fengting Su, Andreas Koeberle

Subject:

Subject ID:	SU003381
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606

Factors:

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

mb_sample_id	local_sample_id	Sample source	Treatment	time point
SA354175	210324_Rescue_Gust_compounds_2h_n2_Ti41_10uM_T58	Breast cancer cells	10 µM Comp. 1	2 h
SA354176	210324_Rescue_Gust_compounds_2h_n1_Ti41_10uM_T46	Breast cancer cells	10 µM Comp. 1	2 h
SA354177	210324_Rescue_Gust_compounds_2h_n3_Ti41_10uM_T70	Breast cancer cells	10 µM Comp. 1	2 h
SA354178	210324_Rescue_Gust_compounds_2h_n1_Ti41_F_10uM_T49	Breast cancer cells	10 µM Comp. 2	2 h
SA354179	210324_Rescue_Gust_compounds_2h_n2_Ti41_F_10uM_T61	Breast cancer cells	10 µM Comp. 2	2 h
SA354180	210324_Rescue_Gust_compounds_2h_n3_Ti41_F_10uM_T73	Breast cancer cells	10 µM Comp. 2	2 h
SA354181	210324_Rescue_Gust_compounds_2h_n3_Ti41_Cl_10uM_T76	Breast cancer cells	10 µM Comp. 3	2 h
SA354182	210324_Rescue_Gust_compounds_2h_n2_Ti41_Cl_10uM_T64	Breast cancer cells	10 µM Comp. 3	2 h
SA354183	210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_10uM_T52	Breast cancer cells	10 µM Comp. 3	2 h
SA354187	210309_MDA_Timecourse_RSL3_n2_24h_RSL3_10uM_24	Breast cancer cells	10 µM RSL3	24 h
SA354188	210309_MDA_Timecourse_RSL3_n3_24h_RSL3_10uM_36	Breast cancer cells	10 µM RSL3	24 h
SA354189	210309_MDA_Timecourse_RSL3_n1_24h_RSL3_10uM_12	Breast cancer cells	10 µM RSL3	24 h
SA354184	210309_MDA_Timecourse_RSL3_n3_2h_RSL3_10uM_27	Breast cancer cells	10 µM RSL3	2 h
SA354185	210309_MDA_Timecourse_RSL3_n2_2h_RSL3_10uM_15	Breast cancer cells	10 µM RSL3	2 h
SA354186	210309_MDA_Timecourse_RSL3_n1_2h_RSL3_10uM_3	Breast cancer cells	10 µM RSL3	2 h
SA354190	210309_MDA_Timecourse_RSL3_n3_4h_RSL3_10uM_30	Breast cancer cells	10 µM RSL3	4 h
SA354191	210309_MDA_Timecourse_RSL3_n2_4h_RSL3_10uM_18	Breast cancer cells	10 µM RSL3	4 h
SA354192	210309_MDA_Timecourse_RSL3_n1_4h_RSL3_10uM_6	Breast cancer cells	10 µM RSL3	4 h
SA354193	210309_MDA_Timecourse_RSL3_n2_6h_RSL3_10uM_21	Breast cancer cells	10 µM RSL3	6 h
SA354194	210309_MDA_Timecourse_RSL3_n3_6h_RSL3_10uM_33	Breast cancer cells	10 µM RSL3	6 h
SA354195	210309_MDA_Timecourse_RSL3_n1_6h_RSL3_10uM_9	Breast cancer cells	10 µM RSL3	6 h
SA354148	210324_Rescue_Gust_compounds_2h_n1_Ti41_1uM_T44	Breast cancer cells	1 µM Comp. 1	2 h
SA354149	210324_Rescue_Gust_compounds_2h_n3_Ti41_1uM_T68	Breast cancer cells	1 µM Comp. 1	2 h
SA354150	210324_Rescue_Gust_compounds_2h_n2_Ti41_1uM_T56	Breast cancer cells	1 µM Comp. 1	2 h
SA354151	210324_Rescue_Gust_compounds_2h_n2_Ti41_F_1uM_T59	Breast cancer cells	1 µM Comp. 2	2 h
SA354152	210324_Rescue_Gust_compounds_2h_n3_Ti41_F_1uM_T71	Breast cancer cells	1 µM Comp. 2	2 h
SA354153	210324_Rescue_Gust_compounds_2h_n1_Ti41_F_1uM_T47	Breast cancer cells	1 µM Comp. 2	2 h
SA354154	210324_Rescue_Gust_compounds_2h_n2_Ti41_Cl_1uM_T62	Breast cancer cells	1 µM Comp. 3	2 h
SA354155	210324_Rescue_Gust_compounds_2h_n3_Ti41_Cl_1uM_T74	Breast cancer cells	1 µM Comp. 3	2 h
SA354156	210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_1uM_T50	Breast cancer cells	1 µM Comp. 3	2 h
SA354157	210324_Rescue_Gust_compounds_2h_n3_RSL3_1uM_Ferrostatin1_3uM_T67	Breast cancer cells	1 µM RSL3 + 3 µM Ferrostatin	2 h
SA354158	210324_Rescue_Gust_compounds_2h_n1_RSL3_1uM_Ferrostatin1_3uM_T43	Breast cancer cells	1 µM RSL3 + 3 µM Ferrostatin	2 h
SA354159	210324_Rescue_Gust_compounds_2h_n2_RSL3_1uM_Ferrostatin1_3uM_T55	Breast cancer cells	1 µM RSL3 + 3 µM Ferrostatin	2 h
SA354166	210309_MDA_Timecourse_RSL3_n3_24h_RSL3_1uM_35	Breast cancer cells	1 µM RSL3	24 h
SA354167	210309_MDA_Timecourse_RSL3_n2_24h_RSL3_1uM_23	Breast cancer cells	1 µM RSL3	24 h
SA354168	210309_MDA_Timecourse_RSL3_n1_24h_RSL3_1uM_11	Breast cancer cells	1 µM RSL3	24 h
SA354160	210324_Rescue_Gust_compounds_2h_n2_RSL3_1uM_T54	Breast cancer cells	1 µM RSL3	2 h
SA354161	210324_Rescue_Gust_compounds_2h_n3_RSL3_1uM_T66	Breast cancer cells	1 µM RSL3	2 h
SA354162	210309_MDA_Timecourse_RSL3_n3_2h_RSL3_1uM_26	Breast cancer cells	1 µM RSL3	2 h
SA354163	210309_MDA_Timecourse_RSL3_n1_2h_RSL3_1uM_2	Breast cancer cells	1 µM RSL3	2 h
SA354164	210324_Rescue_Gust_compounds_2h_n1_RSL3_1uM_T42	Breast cancer cells	1 µM RSL3	2 h
SA354165	210309_MDA_Timecourse_RSL3_n2_2h_RSL3_1uM_14	Breast cancer cells	1 µM RSL3	2 h
SA354169	210309_MDA_Timecourse_RSL3_n3_4h_RSL3_1uM_29	Breast cancer cells	1 µM RSL3	4 h
SA354170	210309_MDA_Timecourse_RSL3_n1_4h_RSL3_1uM_5	Breast cancer cells	1 µM RSL3	4 h
SA354171	210309_MDA_Timecourse_RSL3_n2_4h_RSL3_1uM_17	Breast cancer cells	1 µM RSL3	4 h
SA354172	210309_MDA_Timecourse_RSL3_n3_6h_RSL3_1uM_32	Breast cancer cells	1 µM RSL3	6 h
SA354173	210309_MDA_Timecourse_RSL3_n1_6h_RSL3_1uM_8	Breast cancer cells	1 µM RSL3	6 h
SA354174	210309_MDA_Timecourse_RSL3_n2_6h_RSL3_1uM_20	Breast cancer cells	1 µM RSL3	6 h
SA354196	210324_Rescue_Gust_compounds_2h_n1_Ti41_3uM_T45	Breast cancer cells	3 µM Comp. 1	2 h
SA354197	210324_Rescue_Gust_compounds_2h_n3_Ti41_3uM_T69	Breast cancer cells	3 µM Comp. 1	2 h
SA354198	210324_Rescue_Gust_compounds_2h_n2_Ti41_3uM_T57	Breast cancer cells	3 µM Comp. 1	2 h
SA354199	210324_Rescue_Gust_compounds_2h_n1_Ti41_F_3uM_T48	Breast cancer cells	3 µM Comp. 2	2 h
SA354200	210324_Rescue_Gust_compounds_2h_n3_Ti41_F_3uM_T72	Breast cancer cells	3 µM Comp. 2	2 h
SA354201	210324_Rescue_Gust_compounds_2h_n2_Ti41_F_3uM_T60	Breast cancer cells	3 µM Comp. 2	2 h
SA354202	210324_Rescue_Gust_compounds_2h_n1_Ti41_Cl_3uM_T51	Breast cancer cells	3 µM Comp. 3	2 h
SA354203	210324_Rescue_Gust_compounds_2h_n3_Ti41_Cl_3uM_T75	Breast cancer cells	3 µM Comp. 3	2 h
SA354204	210324_Rescue_Gust_compounds_2h_n2_Ti41_Cl_3uM_T63	Breast cancer cells	3 µM Comp. 3	2 h
SA354211	210309_MDA_Timecourse_RSL3_n3_24h_DMSO_34	Breast cancer cells	DMSO	24 h
SA354212	210309_MDA_Timecourse_RSL3_n2_24h_DMSO_22	Breast cancer cells	DMSO	24 h
SA354213	210309_MDA_Timecourse_RSL3_n1_24h_DMSO_10	Breast cancer cells	DMSO	24 h
SA354205	210324_Rescue_Gust_compounds_2h_n3_DMSO_T65	Breast cancer cells	DMSO	2 h
SA354206	210309_MDA_Timecourse_RSL3_n1_2h_DMSO_1	Breast cancer cells	DMSO	2 h
SA354207	210324_Rescue_Gust_compounds_2h_n2_DMSO_T53	Breast cancer cells	DMSO	2 h
SA354208	210309_MDA_Timecourse_RSL3_n3_2h_DMSO_25	Breast cancer cells	DMSO	2 h
SA354209	210309_MDA_Timecourse_RSL3_n2_2h_DMSO_13	Breast cancer cells	DMSO	2 h
SA354210	210324_Rescue_Gust_compounds_2h_n1_DMSO_T41	Breast cancer cells	DMSO	2 h
SA354214	210309_MDA_Timecourse_RSL3_n3_4h_DMSO_28	Breast cancer cells	DMSO	4 h
SA354215	210309_MDA_Timecourse_RSL3_n2_4h_DMSO_16	Breast cancer cells	DMSO	4 h
SA354216	210309_MDA_Timecourse_RSL3_n1_4h_DMSO_4	Breast cancer cells	DMSO	4 h
SA354217	210309_MDA_Timecourse_RSL3_n3_6h_DMSO_31	Breast cancer cells	DMSO	6 h
SA354218	210309_MDA_Timecourse_RSL3_n2_6h_DMSO_19	Breast cancer cells	DMSO	6 h
SA354219	210309_MDA_Timecourse_RSL3_n1_6h_DMSO_7	Breast cancer cells	DMSO	6 h

Showing results 1 to 72 of 72

Showing results 1 to 72 of 72

Collection:

Collection ID:	CO003374
Collection Summary:	Cultured cells were washed, trypsinized, counted and flash-frozen in liquid N2 and stored at -80°C.
Sample Type:	Breast cancer cells
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR003390
Treatment Summary:	Treatment of brest cancer cells (MDA-MB-231 cells) for the analysis of oxidized phosphatidylethanolamine (oxPE), oxidized phosphatidylcholine (oxPC), and oxidized phosphatidylinositol (oxPI): Human MDA-MB-231 breast cancer cells were treated with vehicle (DMSO) or RSL3 (1 or 10 µM) with or without ferrostatin-1 (3 µM) for 2 h, 4 h, 6 h, or 24 h or with SCs (1, 3, and 10 µM) for 2 h at 37°C and 5% CO2. Cells were harvested, washed with PBS pH 7.4, snap-frozen, and stored at -80°C.

Treatment ID:

TR003390

Treatment Summary:

Treatment of brest cancer cells (MDA-MB-231 cells) for the analysis of oxidized phosphatidylethanolamine (oxPE), oxidized phosphatidylcholine (oxPC), and oxidized phosphatidylinositol (oxPI): Human MDA-MB-231 breast cancer cells were treated with vehicle (DMSO) or RSL3 (1 or 10 µM) with or without ferrostatin-1 (3 µM) for 2 h, 4 h, 6 h, or 24 h or with SCs (1, 3, and 10 µM) for 2 h at 37°C and 5% CO2. Cells were harvested, washed with PBS pH 7.4, snap-frozen, and stored at -80°C.

Sample Preparation:

Sampleprep ID:	SP003388
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℃

Combined analysis:

Analysis ID	AN005345
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	absolute intensities

Chromatography:

Chromatography ID:	CH004047
Chromatography Summary:	Chromatographic separation of phospholipids was carried out on an Acquity BEH C8 column (1.7 μm, 130 Å, 2.1×100 mm, Waters, Milford, MA) using an ExionLC UHPLC system.
Instrument Name:	Waters Acquity H-Class
Column Name:	Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um)
Column Temperature:	45°C
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:	water/acetonitrile 90/10, 2 mM ammonium acetate
Solvent B:	water/acetonitrile 5/95, 2 mM ammonium acetate
Chromatography Type:	Reversed phase

MS:

MS ID:	MS005075
Analysis ID:	AN005345
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). Oxidized phospholipid anions (PE and PI: [M-H]-; PC: [M+OAc]-) were detected after fragmentation to both fatty acid anions. Retention time windows for oxidized PC and oxidized PE were predicted based on the analysis of oxidized PC(16:0/20:4) (oxPAPC) (Avanti Polar Lipids, Alabaster, AL; 1[O]: 2.8–4.6 min; 2[O]: 2.9–4.3 min; 3[O]: 1.45–2.3 min). Quantitation is based on the most intense signals of oxidized fatty acid anion fragments. The fractions of phospholipids with one 1[O], two 2[O], or three oxygens 3[O] incorporated comprise multiple isomeric species that were summed and normalized to DMPC (for oxidized PC and oxidized PI) or DMPE (for oxidized PE) and cell number.
Ion Mode:	NEGATIVE