Summary of Study ST003876

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 PR002430. The data can be accessed directly via it's Project DOI: 10.21228/M8J53Q 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 IDST003876
Study TitleExploration of AGMO-dependent changes in the lipid mediator profile of polarized BMDM cells (M0, M1 and M2 phenoptypes)
Study SummaryMurine BMDMs (Bone marrow-derived macrophages) isolated from wildtype (WT) or AGMO (alkylglycerol monooxygenase) knockout (KO) mice were polarized to the M0, M1 or M2 phenotypes, and then incubated in PBS + Ca2+ (1 mM) for 3 h. Lipid mediators were extracted and analyzed by UPLC-MS/MS.
Institute
University of Innsbruck
Last NameRao
First NameZhigang
AddressMitterweg 24, Innsbruck, Tyrol, 6020, Austria
Emailrzgwuda@gmail.com
Phone+43 662 8044 5581
Submit Date2025-04-17
Num Groups2 (wild type and AGMO knockout)
Total Subjects17
Num Males9
Num Females8
Publicationsin revision
Raw Data AvailableYes
Raw Data File Type(s)mzML, wiff
Analysis Type DetailLC-MS
Release Date2025-04-28
Release Version1
Zhigang Rao Zhigang Rao
https://dx.doi.org/10.21228/M8J53Q
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002430
Project DOI:doi: 10.21228/M8J53Q
Project Title:Alkyglycerol monooxygenase represses prostanoid biosynthesis in a sex-dependent manner
Project Summary:Ether lipids are major constituents of biomembranes and often contain polyunsaturated fatty acids at the sn-2 position, implicating them in cellular signaling and inflammatory processes. Alkylglycerol monooxygenase is the only enzyme capable of cleaving alkylglycerols, an ether lipid subclass, but its exact role and that of its substrates remain unclear. Here, we demonstrate a sex- and cell type-dependent role of alkylglycerol monooxygenase in limiting prostanoid formation without affecting polyunsaturated fatty acid release, as revealed by metabololipidomics profiling. This female-specific effect is driven by the suppression of prostaglandin G/H synthase 2 transcription, as deficiency in alkylglycerol monooxygenase significantly elevated prostaglandin G/H synthase 2 expression in female bone marrow-derived macrophages of the M1 phenotype. Furthermore, this regulatory role of alkylglycerol monooxygenase extends to visceral white adipose tissue, where elevated prostaglandin G/H synthase 2 expression and enhanced prostaglandin production were observed in female samples following alkylglycerol monooxygenase knockout. Collectively, our results expand the immunomodulatory functions of ether lipid metabolism and highlight the role of alkylglycerol monooxygenase in controlling lipid mediator production and maintaining tissue homeostasis.
Institute:University of Innsbruck
Department:Michael Popp Institute
Last Name:Rao
First Name:Zhigang
Address:Mitetrweg 24, Innsbruck, Tyrol, 6020, Austria
Email:rzgwuda@gmail.com
Phone:+43 662 8044 5581
Publications:in revision
Contributors:Zhigang Rao, Andreas Koeberle

Subject:

Subject ID:SU004011
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

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

mb_sample_id local_sample_id Genotype Treatment
SA426432210512_BMDM_0430 M0_TG1 vehAGMO knockout -
SA426433210709_BMDM_male_M0_TG_veh_AGMO_Rao_2AGMO knockout -
SA426434210709_BMDM_male_M1_TG_veh_AGMO_Rao_4AGMO knockout -
SA426435210709_BMDM_male_M2_TG_veh_AGMO_Rao_6AGMO knockout -
SA426436210512_BMDM_0415 M0_TG1 vehAGMO knockout -
SA426437210805_BMDM_male_M0_TG_veh_AGMO_Rao_2AGMO knockout -
SA426438210805_BMDM_male_M1_TG_veh_AGMO_Rao_4AGMO knockout -
SA426439210805_BMDM_male_M2_TG_veh_AGMO_Rao_6AGMO knockout -
SA426440210805_BMDM_male_M0_TG_veh_AGMO_Rao_8AGMO knockout -
SA426441210805_BMDM_male_M1_TG_veh_AGMO_Rao_10AGMO knockout -
SA426442210805_BMDM_male_M2_TG_veh_AGMO_Rao_12AGMO knockout -
SA426443210512_BMDM_0430 M2_TG2 vehAGMO knockout -
SA426444210512_BMDM_0430 M1_TG2 vehAGMO knockout -
SA426445210512_BMDM_0430 M0_TG2 vehAGMO knockout -
SA426446210512_BMDM_0430 M2_TG1 vehAGMO knockout -
SA426447210512_BMDM_0430 M1_TG1 vehAGMO knockout -
SA426448211222_BMDM_AGMO_male_M2_TG1 veh_ 9AGMO knockout -
SA426449210512_BMDM_0415 M0_TG2 vehAGMO knockout -
SA426450211222_BMDM_AGMO_male_M1_TG1 veh_ 6AGMO knockout -
SA426451210512_BMDM_0415 M2_TG2 vehAGMO knockout -
SA426452211222_BMDM_AGMO_male_M0_TG1 veh_ 3AGMO knockout -
SA426453210512_BMDM_0415 M1_TG1 vehAGMO knockout -
SA426454210512_BMDM_0415 M2_TG1 vehAGMO knockout -
SA426455210512_BMDM_0415 M1_TG2 vehAGMO knockout -
SA426456210709_BMDM_male_M2_TG_A23187_AGMO_Rao_24AGMO knockout A23187
SA426457211222_BMDM_AGMO_male_M0_TG1 A23187_ 21AGMO knockout A23187
SA426458211222_BMDM_AGMO_male_M1_TG1 A23187_ 24AGMO knockout A23187
SA426459210805_BMDM_male_M1_TG_A23187_AGMO_Rao_34AGMO knockout A23187
SA426460210709_BMDM_male_M1_TG_A23187_AGMO_Rao_22AGMO knockout A23187
SA426461210805_BMDM_male_M0_TG_A23187_AGMO_Rao_32AGMO knockout A23187
SA426462210512_BMDM_0430 M0_TG1 A23187AGMO knockout A23187
SA426463210512_BMDM_0430 M1_TG1 A23187AGMO knockout A23187
SA426464211222_BMDM_AGMO_male_M2_TG1 A23187_ 27AGMO knockout A23187
SA426465210512_BMDM_0430 M2_TG1 A23187AGMO knockout A23187
SA426466210512_BMDM_0430 M0_TG2 A23187AGMO knockout A23187
SA426467210709_BMDM_male_M0_TG_A23187_AGMO_Rao_20AGMO knockout A23187
SA426468210512_BMDM_0430 M2_TG2 A23187AGMO knockout A23187
SA426469210512_BMDM_0415 M2_TG2 A23187AGMO knockout A23187
SA426470210512_BMDM_0415 M0_TG1 A23187AGMO knockout A23187
SA426471210512_BMDM_0415 M0_TG2 A23187AGMO knockout A23187
SA426472210805_BMDM_male_M2_TG_A23187_AGMO_Rao_30AGMO knockout A23187
SA426473210512_BMDM_0415 M1_TG1 A23187AGMO knockout A23187
SA426474210512_BMDM_0415 M1_TG2 A23187AGMO knockout A23187
SA426475210512_BMDM_0430 M1_TG2 A23187AGMO knockout A23187
SA426476210805_BMDM_male_M1_TG_A23187_AGMO_Rao_28AGMO knockout A23187
SA426477210805_BMDM_male_M2_TG_A23187_AGMO_Rao_36AGMO knockout A23187
SA426478210805_BMDM_male_M0_TG_A23187_AGMO_Rao_26AGMO knockout A23187
SA426479210512_BMDM_0415 M2_TG1 A23187AGMO knockout A23187
SA426480210709_BMDM_male_M0_TG_SACM_AGMO_Rao_11AGMO knockout SACM
SA426481210805_BMDM_male_M1_TG_SACM_AGMO_Rao_16AGMO knockout SACM
SA426482210709_BMDM_male_M1_TG_SACM_AGMO_Rao_13AGMO knockout SACM
SA426483210805_BMDM_male_M0_TG_SACM_AGMO_Rao_14AGMO knockout SACM
SA426484210805_BMDM_male_M2_TG_SACM_AGMO_Rao_24AGMO knockout SACM
SA426485210805_BMDM_male_M1_TG_SACM_AGMO_Rao_22AGMO knockout SACM
SA426486210805_BMDM_male_M0_TG_SACM_AGMO_Rao_20AGMO knockout SACM
SA426487210805_BMDM_male_M2_TG_SACM_AGMO_Rao_18AGMO knockout SACM
SA426488211222_BMDM_AGMO_male_M2_TG1 SACM_ 18AGMO knockout SACM
SA426489211222_BMDM_AGMO_male_M1_TG1 SACM_ 15AGMO knockout SACM
SA426490211222_BMDM_AGMO_male_M0_TG1 SACM_ 12AGMO knockout SACM
SA426491210709_BMDM_male_M2_TG_SACM_AGMO_Rao_15AGMO knockout SACM
SA426492210512_BMDM_0415 M2_TG1 SACMAGMO knockout SACM
SA426493210512_BMDM_0430 M0_TG2 SACMAGMO knockout SACM
SA426494210512_BMDM_0415 M2_TG2 SACMAGMO knockout SACM
SA426495210512_BMDM_0430 M2_TG1 SACMAGMO knockout SACM
SA426496210512_BMDM_0430 M2_TG2 SACMAGMO knockout SACM
SA426497210512_BMDM_0430 M1_TG2 SACMAGMO knockout SACM
SA426498210512_BMDM_0430 M0_TG1 SACMAGMO knockout SACM
SA426499210512_BMDM_0415 M1_TG2 SACMAGMO knockout SACM
SA426500210512_BMDM_0415 M1_TG1 SACMAGMO knockout SACM
SA426501210512_BMDM_0415 M0_TG2 SACMAGMO knockout SACM
SA426502210512_BMDM_0415 M0_TG1 SACMAGMO knockout SACM
SA426503210512_BMDM_0430 M1_TG1 SACMAGMO knockout SACM
SA426504210805_BMDM_male_M1_WT_veh_AGMO_Rao_9wildtype -
SA426505210512_BMDM_0430 M2_WT vehwildtype -
SA426506210805_BMDM_male_M2_WT_veh_AGMO_Rao_11wildtype -
SA426507210512_BMDM_0430 M2_WT2 vehwildtype -
SA426508210805_BMDM_male_M0_WT_veh_AGMO_Rao_7wildtype -
SA426509210512_BMDM_0430 M0_WT2 vehwildtype -
SA426510210512_BMDM_0415 M0_WT vehwildtype -
SA426511210512_BMDM_0430 M1_WT vehwildtype -
SA426512210512_BMDM_0430 M0_WT1 vehwildtype -
SA426513211222_BMDM_AGMO_male_M0_WT1 veh_ 1wildtype -
SA426514211222_BMDM_AGMO_male_M0_WT2 veh_ 2wildtype -
SA426515211222_BMDM_AGMO_male_M1_WT1 veh_ 4wildtype -
SA426516211222_BMDM_AGMO_male_M1_WT2 veh_ 5wildtype -
SA426517211222_BMDM_AGMO_male_M2_WT1 veh_ 7wildtype -
SA426518211222_BMDM_AGMO_male_M2_WT2 veh_ 8wildtype -
SA426519210512_BMDM_0415 M2_WT vehwildtype -
SA426520210512_BMDM_0415 M1_WT vehwildtype -
SA426521210805_BMDM_male_M2_WT_veh_AGMO_Rao_5wildtype -
SA426522210512_BMDM_0430 M1_WT2 vehwildtype -
SA426523210805_BMDM_male_M1_WT_veh_AGMO_Rao_3wildtype -
SA426524210709_BMDM_male_M2_WT_veh_AGMO_Rao_5wildtype -
SA426525210805_BMDM_male_M0_WT_veh_AGMO_Rao_1wildtype -
SA426526210709_BMDM_male_M0_WT_veh_AGMO_Rao_1wildtype -
SA426527210709_BMDM_female_M2_WT_veh_AGMO_Rao_9wildtype -
SA426528210709_BMDM_female_M1_WT_veh_AGMO_Rao_8wildtype -
SA426529210709_BMDM_male_M1_WT_veh_AGMO_Rao_3wildtype -
SA426530210709_BMDM_female_M0_WT_veh_AGMO_Rao_7wildtype -
SA426531210805_BMDM_male_M0_WT_A23187_AGMO_Rao_31wildtype A23187
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Collection:

Collection ID:CO004004
Collection Summary:Murine BMDMs were isolated from wildtype or Agmo knockout mice (9 female, 8 male mice) and then kept in culture in a humidified atmosphere at 37 °C with 5% CO2. After polarization into M0, M1 or M2 phenotypes, cells were treated and supernatants (172 samples) were collected in glass tubes containing ice-cold methanol (on ice), stored at -20 °C for more than 2 h to allow protein precipitation. After centrifugation (1200 × g, 10 min, 4 °C), supernatants were transferred to new glass tubes and mixed with 8 mL of ice-cold acidified water (pH = 3.5), followed by solid phase extraction (SPE). SPE eluents were collected and brought to dryness using a TurboVap system (TurboVap LV, Biotage, Uppsala, Sweden) under nitrogen flow at 30 °C. Then the dried lipid film was dissolved in methanol/water (50/50, v/v), stored at -80 °C before subjection to lipid mediator analysis by UPLC-MS/MS.
Sample Type:Macrophages
Storage Conditions:-80℃

Treatment:

Treatment ID:TR004020
Treatment Summary:Murine BMDMs isolated from wildtype or Agmo knockout mice were polarized to the M0, M1 or M2 phenotypes, and then stimulated with Staphylococcus aureus-conditioned medium (SACM, 1%, 3 h) or A23187 (2.5 µM, 15 min) in PBS + Ca2+ (1 mM) for the indicated time.

Sample Preparation:

Sampleprep ID:SP004017
Sampleprep Summary:Cell supernatants (1.5 mL) or tissue homogenates (150 µL) were mixed with ice-cold methanol (3 mL) containing the deuterated internal standards d8-5S-HETE, d4-LTB4, d5-LXA4, d5-RvD2, d4-PGE2 (20 pg/µL, 10 µL each, Cayman Chemicals, Ann Arbor, MI) and d8-arachidonic acid (200 pg/µL, 10 µL, Cayman Chemicals). The sample mixtures were first kept at -20°C for more than 2 h to allow protein precipitation, and then centrifuged (1200 × g, 10 min, 4°C). Supernatants were transferred to new vials and mixed with 8 mL of acidified water (pH = 3.5) before loading onto Sep-Pak C18 6cc Vac Cartridges (500 mg; Waters, Milford, MA) (Neukirch et al. 2021) pre-equilibrated with 6 mL methanol and 2 mL water. After several washing steps with water (6 mL) and n-hexane (6 mL), lipid mediators were eluted with methyl formate. The eluate was brought to dryness using a TurboVap system (TurboVap LV, Biotage, Uppsala, Sweden) under nitrogen flow at 30°C. The dried lipid film was dissolved in methanol/water (50/50, v/v) and analyzed by ultra-performance liquid chromatography‒tandem mass spectrometry (UPLC-MS/MS).
Processing Storage Conditions:-20℃
Extract Storage:-80℃

Chromatography:

Chromatography ID:CH004831
Chromatography Summary:Chromatographic separation of phospholipids was carried out on an Acquity BEH C18 column (ACQUITY UPLC® BEH C18; 1.7 µm; 2.1 mm × 100 mm; Waters) using an Exion LC system (Sciex, Darmstadt, Germany).
Instrument Name:Waters Acquity H-Class
Column Name:Waters ACQUITY UPLC BEH C18 (100 x 2.1 mm, 1.7 µm)
Column Temperature:55°C
Flow Gradient:The gradient was ramped from 35.6% B to 84.4% B over 12.5 min, then to 97.8% B and maintained for another 5 min, and finally isocratically set to 35.6% B for 2.5 min.
Flow Rate:0.35 mL/min
Solvent A:90% water/10% Methanol; 0.01% Acetic acid
Solvent B:100% Methanol; 0.01% Acetic acid
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN006368
Analysis Type:MS
Chromatography ID:CH004831
Num Factors:6
Num Metabolites:29
Units:absolute intensity (pg)
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