Summary of Study ST001788

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 PR001029. The data can be accessed directly via it's Project DOI: 10.21228/M8PM56 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 IDST001788
Study Titleβ-Adrenergic regulation of metabolism in macrophages (part-IV)
Study SummaryMacrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. β-adrenoceptor (βAR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of β-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that βAR signalling has on the metabolome and proteome. We found that βAR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that βAR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that βAR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that βAR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
Institute
Monash University
Last NamePeterson
First NameAmanda
AddressDrug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Emailamanda.peterson@monash.edu
Phone99039282
Submit Date2021-05-13
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2021-05-28
Release Version1
Amanda Peterson Amanda Peterson
https://dx.doi.org/10.21228/M8PM56
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001029
Project DOI:doi: 10.21228/M8PM56
Project Title:β-Adrenergic regulation of metabolism in macrophages
Project Summary:Macrophages have important roles in the immune system including clearing pathogens and wound healing. Metabolic phenotypes have been associated with functional phenotypes, where pro-inflammatory macrophages have an increased rate of glycolysis and anti-inflammatory macrophages primarily use oxidative phosphorylation. β-adrenoceptor (βAR) signalling in macrophages has been implicated in disease states such as cancer, atherosclerosis and rheumatoid arthritis. The impact of β-adrenoceptor signalling on macrophage metabolism has not been defined. Here we expand on defining the phenotype of macrophages treated with isoprenaline and describe the impact that βAR signalling has on the metabolome and proteome. We found that βAR signalling alters proteins involved in cytoskeletal rearrangement and redox control of the cell. We showed that βAR signalling in macrophages shifts glucose metabolism from glycolysis towards the tricarboxylic acid cycle and pentose phosphate pathways. We also show that βAR signalling perturbs purine metabolism by accumulating adenylate pools. Taken together these results indicate that βAR signalling shifts metabolism to support redox perturbations and upregulate proteins involved in cytoskeletal changes that may impact migration and phagocytosis processes.
Institute:Monash University
Last Name:Peterson
First Name:Amanda
Address:Drug delivery, disposition and dynamics, Pharmacy and Pharmaceutical Sciences, 381 Royal Parade, Parkville, Victoria, 3052, Australia
Email:amanda.peterson@monash.edu
Phone:99039282

Subject:

Subject ID:SU001865
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id treatment
SA166492M_labcontrol_2Control_13C-Glucose_4hr
SA166493M_labcontrol_3Control_13C-Glucose_4hr
SA166494M_labcontrol_4Control_13C-Glucose_4hr
SA166495CP_labcontrol_1Control_13C-Glucose_4hr
SA166496M_labcontrol_1Control_13C-Glucose_4hr
SA166497CP_labcontrol_2Control_13C-Glucose_4hr
SA166498CP_labcontrol_4Control_13C-Glucose_4hr
SA166499CP_labcontrol_3Control_13C-Glucose_4hr
SA166500M_labisoprenaline_3Isoprenaline_13C-Glucose_4hr
SA166501M_labisoprenaline_4Isoprenaline_13C-Glucose_4hr
SA166502CP_labisoprenaline_4Isoprenaline_13C-Glucose_4hr
SA166503M_labisoprenaline_2Isoprenaline_13C-Glucose_4hr
SA166504M_labisoprenaline_1Isoprenaline_13C-Glucose_4hr
SA166505CP_labisoprenaline_2Isoprenaline_13C-Glucose_4hr
SA166506CP_labisoprenaline_3Isoprenaline_13C-Glucose_4hr
SA166507CP_labisoprenaline_1Isoprenaline_13C-Glucose_4hr
Showing results 1 to 16 of 16

Collection:

Collection ID:CO001858
Collection Summary:500 μL of culture medium was collected and centrifuged for 5 minutes at 200 x g at 4°C. 20 μL of the supernatant is transferred to a new Eppendorf tube where 180 μL of 50:50, methanol: acetonitrile (ACN) containing 1 μM internal standards (CHAPS, CAPS, PIPES and TRIS) is added. The samples are placed on a shaker for 30 minutes. Samples were then vortexed for 30 seconds before centrifuging at 20,000 x g for 10 minutes at 4°C. 160 μL of supernatant was transferred to a plastic LC-MS vial and stored at -80°C until LC-MS analysis.
Sample Type:Macrophages

Treatment:

Treatment ID:TR001878
Treatment Summary:Differentiated macrophage-like cells (9.9x106) were seeded onto fibronectin-coated glass dishes. After 24 hours in culture, cells were treated with 1 µM isoprenaline (or vehicle) for 20 hours and 11 mM U-13C6-D-Glucose labelled medium was spiked in for the final 4 hours (to give a final ratio of 50:50 of U-13C and U-12C D-glucose).

Sample Preparation:

Sampleprep ID:SP001871
Sampleprep Summary:500 μL of culture medium was collected and centrifuged for 5 minutes at 200 x g at 4°C. 20 μL of the supernatant is transferred to a new Eppendorf tube where 180 μL of 50:50, methanol: acetonitrile (ACN) containing 1 μM internal standards (CHAPS, CAPS, PIPES and TRIS) is added. The samples are placed on a shaker for 30 minutes. Samples were then vortexed for 30 seconds before centrifuging at 20,000 x g for 10 minutes at 4°C. 160 μL of supernatant was transferred to a plastic LC-MS vial and stored at -80°C until LC-MS analysis.

Chromatography:

Chromatography ID:CH002150
Instrument Name:Thermo Dionex Ultimate 3000
Column Name:ZIC-pHILIC (150 x 4.6mm,5um)
Chromatography Type:HILIC

Analysis:

Analysis ID:AN002899
Analysis Type:MS
Chromatography ID:CH002150
Num Factors:2
Num Metabolites:556
Rt Units:Minutes
Units:Intensity
  
Analysis ID:AN002900
Analysis Type:MS
Chromatography ID:CH002150
Num Factors:2
Num Metabolites:557
Rt Units:Minutes
Units:Intensity
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