Summary of Study ST001124

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 PR000732. The data can be accessed directly via it's Project DOI: 10.21228/M82D79 This work is supported by NIH grant, U2C- DK119886.

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Study IDST001124
Study TitleEarly Mechanistic Events Induced by Secondhand Smoke Prevalent Low Molecular Weight Polycyclic Aromatic Hydrocarbons in Mouse Lung Epithelial Cells
Study SummaryWe evaluated lung epithelial cells exposed to low molecular weight polycyclic aromatic hydrocarbons and what lipid metabolites were produced following early exposure, prior to metabolism. For the targeted study (AKB study 2, we used 40uM dose of the 2PAHs (1methylanthracene and fluoranthene; 1:1 ratio)and assessed 2,4,8,and 12 hrs of treatment with the PAHs. We also examined a 24 h time point in another study at a lower dose (15 uM LMW PAH mixture; 1:1 ratio of 1-methanthrancene and fluoranthene).
Institute
University of Colorado Denver
DepartmentAnschutz Medical Campus
Last NameBauer
First NameAlison
Address12850 E. Montview Dr. Rm 3125, Aurora, CO 80045
Emailalison.bauer@ucdenver.edu
Phone303-724-6297
Submit Date2018-11-06
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2019-02-27
Release Version1
Alison Bauer Alison Bauer
https://dx.doi.org/10.21228/M82D79
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR000732
Project DOI:doi: 10.21228/M82D79
Project Title:Adverse effects of PAHs on lung cells
Project Summary:Low-molecular-weight (LMW) polycyclic aromatic hydrocarbons (PAHs) are more prevalent in the environment, occupational settings, as well as in secondhand smoke (SHS), when compared to their high molecular weight counterparts, such as benzo[a]pyrene (B[a]P). Previously, we demonstrated that SHS-prevalent LMW PAHs activate p38-MAPK-dependent dysregulation of gap junction intercellular communication (GJIC) and increased cytokines involved in inflammatory lung diseases. However, there is little known about the early mechanistic events leading to inflammation, specifically those mediated through lipid signaling and eicosanoids. Secondhand smoke is a complex mixture and to model this feature in vitro we examined the effects of a binary mixture of 1-methylanthracene (1-MeA) and fluoranthene (Flthn) in C10 cells, a mouse, non-tumorigenic alveolar type II cell line via a global metabolomics approach to evaluate the lipids.
Institute:University of Colorado Denver
Department:Pharmaceutical Sciences, Anschutz Medical Campus
Last Name:Bauer
First Name:Alison
Address:12850 E. Montview Dr. Rm 3125, Aurora, CO 80045
Email:alison.bauer@ucdenver.edu
Phone:303-724-6297
Funding Source:R15 ES 024893-01 (AKB) and the Flight Attendant Medical Research Institute (FAMRI) CIA 130022 (AKB).
Project Comments:There are two studies that will be uploaded. AKB study 1 and AKB study 2.
Contributors:Katelyn J. Siegrist, DeeDee Romo, Brad L. Upham, Michael Armstrong, Kevin Quinn, Lauren Vanderlinden, Kalpana Velmurugan, Mark Elie, Dominik Reinhold, Nichole Reisdorph, Laura Saba

Subject:

Subject ID:SU001185
Subject Type:Cultured cells
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

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

mb_sample_id local_sample_id Treatment Time
SA077960592PAHS+p38inhibitor 12
SA077961602PAHS+p38inhibitor 12
SA077962582PAHS+p38inhibitor 12
SA077963572PAHS+p38inhibitor 12
SA07796492PAHS+p38inhibitor 2
SA077965112PAHS+p38inhibitor 2
SA077966122PAHS+p38inhibitor 2
SA077967102PAHS+p38inhibitor 2
SA077968762PAHS+p38inhibitor 24
SA077969742PAHS+p38inhibitor 24
SA077970752PAHS+p38inhibitor 24
SA077971732PAHS+p38inhibitor 24
SA077972272PAHS+p38inhibitor 4
SA077973262PAHS+p38inhibitor 4
SA077974252PAHS+p38inhibitor 4
SA077975282PAHS+p38inhibitor 4
SA077976442PAHS+p38inhibitor 8
SA077977432PAHS+p38inhibitor 8
SA077978422PAHS+p38inhibitor 8
SA077979412PAHS+p38inhibitor 8
SA077980502PAHs 12
SA077981522PAHs 12
SA077982492PAHs 12
SA077983512PAHs 12
SA07798432PAHs 2
SA07798512PAHs 2
SA07798622PAHs 2
SA07798742PAHs 2
SA077988652PAHs 24
SA077989672PAHs 24
SA077990682PAHs 24
SA077991662PAHs 24
SA077992202PAHs 4
SA077993172PAHs 4
SA077994182PAHs 4
SA077995192PAHs 4
SA077996352PAHs 8
SA077997332PAHs 8
SA077998362PAHs 8
SA077999342PAHs 8
SA07800054DMSO 12
SA07800155DMSO 12
SA07800256DMSO 12
SA07800353DMSO 12
SA0780048DMSO 2
SA0780056DMSO 2
SA0780067DMSO 2
SA0780075DMSO 2
SA07800870DMSO 24
SA07800969DMSO 24
SA07801072DMSO 24
SA07801171DMSO 24
SA07801222DMSO 4
SA07801321DMSO 4
SA07801423DMSO 4
SA07801524DMSO 4
SA07801639DMSO 8
SA07801738DMSO 8
SA07801837DMSO 8
SA07801940DMSO 8
SA07802064p38 inhibitor 12
SA07802163p38 inhibitor 12
SA07802261p38 inhibitor 12
SA07802362p38 inhibitor 12
SA07802415p38 inhibitor 2
SA07802514p38 inhibitor 2
SA07802616p38 inhibitor 2
SA07802713p38 inhibitor 2
SA07802877p38 inhibitor 24
SA07802979p38 inhibitor 24
SA07803080p38 inhibitor 24
SA07803178p38 inhibitor 24
SA07803230p38 inhibitor 4
SA07803331p38 inhibitor 4
SA07803429p38 inhibitor 4
SA07803532p38 inhibitor 4
SA07803645p38 inhibitor 8
SA07803746p38 inhibitor 8
SA07803848p38 inhibitor 8
SA07803947p38 inhibitor 8
Showing results 1 to 80 of 80

Collection:

Collection ID:CO001179
Collection Summary:Immediately following treatment, cell monolayers were washed 3 times with cold PBS, and cells were scraped and fixed in 500 μL 70% MeOH. Samples were stored immediately at -80°C until pretreatment for solid phase extraction (SPE), as follows.
Collection Protocol Filename:AKB_Targeted-protocol.docx
Collection Protocol Comments:For AKB study 2 protocol, the collection, treatment, and sample prep are all in the same file.
Sample Type:Epithelial cells

Treatment:

Treatment ID:TR001200
Treatment Summary:The C10 cell line was obtained from Dr. Lori Nield (University of Colorado). These cells are an immortalized, non-transformed alveolar type II cell line originally derived from a BALB mouse (Bentel et al. 1989; Malkinson et al. 1997). C10 cells were treated with or without p38 inhibitor (CAY10502) 24 h following serum deprivation with either DMSO control or 40 μM the binary LMW PAH mixture for 2, 4, 8, 12, or 24 hr and DMSO control with an n=4/treatment.
Treatment Protocol Filename:AKB_Targeted-protocol.docx
Treatment Protocol Comments:For AKB study 2 protocol, the collection, treatment, and sample prep are all in the same file.

Sample Preparation:

Sampleprep ID:SP001193
Sampleprep Summary:Briefly, before SPE cleanup, the frozen samples were removed from the freezer and centrifuged at 14,000 RPM at 4°C for 10 minutes. The supernatant was removed, transferred to a new microcentrifuge tube, and spiked with 10 μL of 10 pg/μL internal standard solution (100 pg of each: 5(S)-HETE-d8, 8-iso-PGF2a-d4, 9(S)-HODE-d4, LTB4-d4, LTD4-d5, LTE4-d5, PGE2-d4, PGF2a-d9 and RvD2-d5 in ethanol). The supernatant was dried in a vacuum centrifuge and reconstituted with 1.0 mL of 10% methanol. The reconstituted extracts were loaded on a Strata-X 33 μm 30 mg/1maL SPE column (Phenomenex, Torrance, California, USA) preconditioned with 1.0 mL of methanol followed by 1.0 mL of water. The SPE column was washed with 10% methanol and then eluted directly into a reduced surface activity/maximum recovery glass autosampler vial with 1.0 mL of methyl formate. The methyl formate was evaporated completely from the vial with a stream of nitrogen and the SPE cartridge was then eluted with 1.0 mL of methanol directly into the same autosampler vial. The methanol was evaporated to dryness with a stream of nitrogen and the sample was reconstituted with 20 uL of ethanol. The samples were analyzed immediately or frozen at -70 °C until analysis.
Sampleprep Protocol Filename:AKB_Targeted-protocol.docx
Sampleprep Protocol Comments:For AKB study 2 protocol, the collection, treatment, and sample prep are all in the same file.

Combined analysis:

Analysis ID AN001849
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1260
Column Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6490 QQQ
Ion Mode NEGATIVE
Units pg per 1.5 million cells

Chromatography:

Chromatography ID:CH001326
Instrument Name:Agilent 1260
Column Name:Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um)
Chromatography Type:Reversed phase

MS:

MS ID:MS001710
Analysis ID:AN001849
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:none
Ion Mode:NEGATIVE
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