Summary of Study ST002548

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 PR001641. The data can be accessed directly via it's Project DOI: 10.21228/M8KM6N 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.

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Study ID	ST002548
Study Title	Equal Flux of Oxidants study
Study Type	untargeted MS
Study Summary	To study the global metabolic impact of myeloperoxidase-derived oxidants on airway epithelial cells, we developed an in vitro oxidant exposure model. Experiments were conducted with BEAS-2B cells that were supplemented with 100pg/ml recombinant epidermal growth factor and 0.1% FBS prior to oxidant exposure. Measurements were taken at 2, and 6 hours for: (i) H2O2, (ii) HOCl, (iii) HOBr, (iv) HOSCN, and (v) control. Additional measurements were taken at 24 hours for HOSCN and control only.
Institute	Emory University
Department	Pediatrics
Laboratory	Joshua Chandler, PhD
Last Name	Chandler
First Name	Joshua
Address	2015 Uppergate Drive, Atlanta, GA 30322
Email	Joshua.chandler@emory.edu
Phone	404-727-3536
Submit Date	2023-01-26
Num Groups	5 (Control, H2O2, HOBr, HOCl, and HOSCN)
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2023-09-19
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001641
Project DOI:	doi: 10.21228/M8KM6N
Project Title:	Myeloperoxidase oxidants elicit distinct metabolomic responses in airway epithelial cells and reveal early cystic fibrosis disease risk factors
Project Summary:	Myeloperoxidase (MPO) is released by neutrophils in inflamed tissues. MPO oxidizes chloride, bromide, and thiocyanate to produce hypochlorous acid (HOCl), hypobromous acid (HOBr), or hypothiocyanous acid (HOSCN), respectively. These oxidants are toxic to pathogens, but they also react with host cells and may elicit redox signaling or cause collateral damage. Here, we investigated how equal dose-rate exposures of MPO-derived oxidants differentially impact the metabolome of human airway epithelial cells (AEC). AECs are important targets of oxidative stress in inflammatory lung diseases such as cystic fibrosis (CF) and chronic obstructive pulmonary disease, in which neutrophils, MPO, and reactive oxygen species are increased compared to healthy individuals. HOSCN-exposed cells were viable after 24 h of continuous oxidant exposure, while HOBr and HOCl exposures were cytotoxic by 24 h. Untargeted metabolomics revealed MPO-derived oxidants caused distinct metabolic effects in AECs. HOCl and HOBr were more similar to each other than either was to HOSCN, and all differed from hydrogen peroxide. We noted methionine oxidation products, methionine sulfoxide and dehydromethionine, were significantly increased compared to controls only in HOCl- and HOBr-exposed cells. Furthermore, the presence of these metabolites in bronchoalveolar lavage fluid from 5 years-olds with CF (n=27) was associated with increased neutrophilic inflammation and bronchiectasis. This indicates that HOCl and/or HOBr are likely present in early CF airways and either contribute to disease progression or associate with a related pathophysiological process. We anticipate this oxidant exposure model can be used to identify additional novel metabolites which could serve as candidate biomarkers of MPO-driven inflammation in human disease.
Institute:	Emory University
Department:	Pediatrics
Laboratory:	Joshua Chandler, PhD
Last Name:	Chandler
First Name:	Joshua
Address:	2015 Uppergate Drive, Atlanta, GA 30322
Email:	Joshua.chandler@emory.edu
Phone:	404-727-3536
Contributors:	Joshua Chandler, Susan Kim

Subject:

Subject ID:	SU002648
Subject Type:	Cultured cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Cell Biosource Or Supplier:	American Type Culture Collection
Cell Strain Details:	BEAS-2B (CRL-9609)

Factors:

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

mb_sample_id	local_sample_id	Treatment	Time
SA255684	20210225_136_C11_Con_24h-neg	Control	24h
SA255685	20210225_135_C11_Con_24h-pos	Control	24h
SA255686	20210228_272_B11_Con_24h-neg	Control	24h
SA255687	20210228_271_B11_Con_24h-pos	Control	24h
SA255688	20210228_268_G11_Con_24h-neg	Control	24h
SA255689	20210225_134_J11_Con_24h-neg	Control	24h
SA255690	20210225_098_E11_Con_24h-neg	Control	24h
SA255691	20210224_084_A11_Con_24h-neg	Control	24h
SA255692	20210224_083_A11_Con_24h-pos	Control	24h
SA255693	20210227_231_D11_Con_24h-pos	Control	24h
SA255694	20210225_097_E11_Con_24h-pos	Control	24h
SA255695	20210228_267_G11_Con_24h-pos	Control	24h
SA255696	20210225_133_J11_Con_24h-pos	Control	24h
SA255697	20210227_232_D11_Con_24h-neg	Control	24h
SA255698	20210226_180_I11_Con_24h-neg	Control	24h
SA255699	20210226_172_F11_Con_24h-neg	Control	24h
SA255700	20210226_171_F11_Con_24h-pos	Control	24h
SA255701	20210228_252_H11_Con_24h-neg	Control	24h
SA255702	20210228_251_H11_Con_24h-pos	Control	24h
SA255703	20210226_179_I11_Con_24h-pos	Control	24h
SA255704	20210228_281_H01_Con_2h-pos	Control	2h
SA255705	20210226_182_D01_Con_2h-neg	Control	2h
SA255706	20210301_306_J01_Con_2h-neg	Control	2h
SA255707	20210228_279_I01_Con_2h-pos	Control	2h
SA255708	20210228_280_I01_Con_2h-neg	Control	2h
SA255709	20210228_282_H01_Con_2h-neg	Control	2h
SA255710	20210301_305_J01_Con_2h-pos	Control	2h
SA255711	20210227_224_G01_Con_2h-neg	Control	2h
SA255712	20210224_085_B01_Con_2h-pos	Control	2h
SA255713	20210224_062_A01_Con_2h-neg	Control	2h
SA255714	20210224_086_B01_Con_2h-neg	Control	2h
SA255715	20210224_087_F01_Con_2h-pos	Control	2h
SA255716	20210301_309_E01_Con_2h-pos	Control	2h
SA255717	20210224_088_F01_Con_2h-neg	Control	2h
SA255718	20210227_223_G01_Con_2h-pos	Control	2h
SA255719	20210301_310_E01_Con_2h-neg	Control	2h
SA255720	20210225_114_J06_Con_2h-neg	Control	2h
SA255721	20210224_061_A01_Con_2h-pos	Control	2h
SA255722	20210228_256_D06_Con_2h-neg	Control	2h
SA255723	20210225_113_J06_Con_2h-pos	Control	2h
SA255724	20210228_255_D06_Con_2h-pos	Control	2h
SA255725	20210226_181_D01_Con_2h-pos	Control	2h
SA255726	20210224_076_F06_Con_6h-neg	Control	6h
SA255727	20210228_266_I06_Con_6h-neg	Control	6h
SA255728	20210224_075_F06_Con_6h-pos	Control	6h
SA255729	20210224_064_E06_Con_6h-neg	Control	6h
SA255730	20210228_265_I06_Con_6h-pos	Control	6h
SA255731	20210224_063_E06_Con_6h-pos	Control	6h
SA255732	20210226_146_B06_Con_6h-neg	Control	6h
SA255733	20210228_259_G06_Con_6h-pos	Control	6h
SA255734	20210225_101_A06_Con_6h-pos	Control	6h
SA255735	20210225_103_C06_Con_6h-pos	Control	6h
SA255736	20210226_176_C01_Con_6h-neg	Control	6h
SA255737	20210226_175_C01_Con_6h-pos	Control	6h
SA255738	20210225_104_C06_Con_6h-neg	Control	6h
SA255739	20210228_260_G06_Con_6h-neg	Control	6h
SA255740	20210226_145_B06_Con_6h-pos	Control	6h
SA255741	20210225_102_A06_Con_6h-neg	Control	6h
SA255742	20210224_056_H06_Con_6h-neg	Control	6h
SA255743	20210224_055_H06_Con_6h-pos	Control	6h
SA255744	20210301_344_QC_Con_ddMS2_GL-CompoundLibrary-neg	Control	n/a
SA255745	20210301_342_QC_Con_ddMS2_EX-neg	Control	n/a
SA255746	20210301_339_QC_Con_ddMS2-pos	Control	n/a
SA255747	20210301_343_QC_Con_ddMS2_GL-CompoundLibrary-pos	Control	n/a
SA255748	20210301_378_QC_Con_ddMS2_SRM-neg	Control	n/a
SA255749	20210301_340_QC_Con_ddMS2-neg	Control	n/a
SA255750	20210301_341_QC_Con_ddMS2_EX-pos	Control	n/a
SA255751	20210301_377_QC_Con_ddMS2_SRM-pos	Control	n/a
SA255752	20210228_250_E02_H2O2_2h-neg	H2O2	2h
SA255753	20210301_299_D07_H2O2_2h-pos	H2O2	2h
SA255754	20210225_123_G02_H2O2_2h-pos	H2O2	2h
SA255755	20210228_244_J07_H2O2_2h-neg	H2O2	2h
SA255756	20210225_124_G02_H2O2_2h-neg	H2O2	2h
SA255757	20210228_249_E02_H2O2_2h-pos	H2O2	2h
SA255758	20210226_162_B02_H2O2_2h-neg	H2O2	2h
SA255759	20210301_311_J02_H2O2_2h-pos	H2O2	2h
SA255760	20210301_312_J02_H2O2_2h-neg	H2O2	2h
SA255761	20210225_119_I02_H2O2_2h-pos	H2O2	2h
SA255762	20210225_120_I02_H2O2_2h-neg	H2O2	2h
SA255763	20210226_161_B02_H2O2_2h-pos	H2O2	2h
SA255764	20210301_300_D07_H2O2_2h-neg	H2O2	2h
SA255765	20210228_243_J07_H2O2_2h-pos	H2O2	2h
SA255766	20210224_074_F02_H2O2_2h-neg	H2O2	2h
SA255767	20210224_073_F02_H2O2_2h-pos	H2O2	2h
SA255768	20210301_296_D02_H2O2_2h-neg	H2O2	2h
SA255769	20210301_295_D02_H2O2_2h-pos	H2O2	2h
SA255770	20210227_233_A02_H2O2_2h-pos	H2O2	2h
SA255771	20210227_234_A02_H2O2_2h-neg	H2O2	2h
SA255772	20210301_297_H02_H2O2_2h-pos	H2O2	2h
SA255773	20210301_298_H02_H2O2_2h-neg	H2O2	2h
SA255774	20210224_059_F07_H2O2_6h-pos	H2O2	6h
SA255775	20210227_204_A07_H2O2_6h-neg	H2O2	6h
SA255776	20210301_308_E07_H2O2_6h-neg	H2O2	6h
SA255777	20210301_307_E07_H2O2_6h-pos	H2O2	6h
SA255778	20210226_150_C02_H2O2_6h-neg	H2O2	6h
SA255779	20210224_060_F07_H2O2_6h-neg	H2O2	6h
SA255780	20210224_079_C07_H2O2_6h-pos	H2O2	6h
SA255781	20210224_080_C07_H2O2_6h-neg	H2O2	6h
SA255782	20210227_203_A07_H2O2_6h-pos	H2O2	6h
SA255783	20210226_149_C02_H2O2_6h-pos	H2O2	6h

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Collection:

Collection ID:	CO002641
Collection Summary:	Cells grown in 60 mm dishes were washed once with 1X PBS before being scraped into 250 ul of 2:2:1 acetonitrile:methanol:water plus 0.1 M formic acid. Then a second 250 ul of PBS was added and pooled with the first to maximize recovery.
Sample Type:	Cultured cells

Treatment:

Treatment ID:	TR002660
Treatment Summary:	Cells were treated with 2 U/L of glucose oxidase (GOX) to produce H2O2. To produce HOCl, 200 U/L of myeloperoxidase (MPO) was added in addition to GOX. To produce HOBr and HOSCN, 1 mM of sodium bromide or sodium thiocyanate were added, respectively, in addition to GOX and MPO. Cells were exposed for up to 2h, 6h, and 24h.

Sample Preparation:

Sampleprep ID:	SP002654
Sampleprep Summary:	Metabolites were extracted from cells using 2:2:1 acetonitrile:methanol:water plus 0.1 M formic acid method. Formic acid was neutralized with ammonium bicarbonate. The samples were incubated at -20C for 30 min then centrifuged at 20,000 g and 4C for 10 min. Supernatants and pellets were stored at -80C. Pellets were later dissolved for total protein assay and the supernatant was normalized for protein concentration. A small, equal volume of each sample was pooled to make a global quality control samples and experimental group-specific quality controls.

Sampleprep ID:

SP002654

Sampleprep Summary:

Metabolites were extracted from cells using 2:2:1 acetonitrile:methanol:water plus 0.1 M formic acid method. Formic acid was neutralized with ammonium bicarbonate. The samples were incubated at -20C for 30 min then centrifuged at 20,000 g and 4C for 10 min. Supernatants and pellets were stored at -80C. Pellets were later dissolved for total protein assay and the supernatant was normalized for protein concentration. A small, equal volume of each sample was pooled to make a global quality control samples and experimental group-specific quality controls.

Combined analysis:

Analysis ID	AN004195	AN004196
Analysis type	MS	MS
Chromatography type	HILIC	HILIC
Chromatography system	Thermo Vanquish	Thermo Vanquish
Column	HILICON iHILIC-(P) Classic (150 x 2.1mm, 5um)	HILICON iHILIC-(P) Classic (150 x 2.1mm, 5um)
MS Type	ESI	ESI
MS instrument type	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive HF hybrid Orbitrap	Thermo Q Exactive HF hybrid Orbitrap
Ion Mode	POSITIVE	NEGATIVE
Units	normalized peak area	normalized peak area

Chromatography:

Chromatography ID:	CH003109
Instrument Name:	Thermo Vanquish
Column Name:	HILICON iHILIC-(P) Classic (150 x 2.1mm, 5um)
Column Temperature:	40
Flow Gradient:	Gradient began at 10% A and progressed to 90% A, followed by a 2-minute hold and 8-minute re-equilibration of the column at 10% A
Flow Rate:	0.2 ml/min
Solvent A:	100% water; 15 mM ammonium acetate, pH 9.4
Solvent B:	100% acetonitrile
Chromatography Type:	HILIC

MS:

MS ID:	MS003942
Analysis ID:	AN004195
Instrument Name:	Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The data acquisition sequence was randomized before injection and a global quality control was injected every 10 samples to determine measurement reproducibility. Data was analyzed using Compound Discoverer 3.3 and FreeStyle 1.7 (Thermo Fisher Scientific). mzCloud (mzcloud.org) and LipidSearch (ThermoFisher) were used for MS/MS identifications
Ion Mode:	POSITIVE

MS ID:	MS003943
Analysis ID:	AN004196
Instrument Name:	Thermo Q Exactive HF hybrid Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The data acquisition sequence was randomized before injection and a global quality control was injected every 10 samples to determine measurement reproducibility. Data was analyzed using Compound Discoverer 3.3 and FreeStyle 1.7 (Thermo Fisher Scientific). mzCloud (mzcloud.org) and LipidSearch (ThermoFisher) were used for MS/MS identifications
Ion Mode:	NEGATIVE