Summary of Study ST001548
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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001548 |
| Study Title | β-Adrenergic regulation of metabolism in macrophages (part-II) |
| Study 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 |
| amanda.peterson@monash.edu | |
| Phone | 99039282 |
| Submit Date | 2020-11-17 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2020-12-01 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
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: | SU001624 |
| 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 |
|---|---|---|
| SA130686 | M_control_1 | Control_13C-Glucose |
| SA130687 | M_control_4 | Control_13C-Glucose |
| SA130688 | M_control_3 | Control_13C-Glucose |
| SA130689 | M_control_2 | Control_13C-Glucose |
| SA130690 | M_isoprenaline_4 | Isoprenaline_13C-Glucose |
| SA130691 | M_isoprenaline_1 | Isoprenaline_13C-Glucose |
| SA130692 | M_isoprenaline_2 | Isoprenaline_13C-Glucose |
| SA130693 | M_isoprenaline_3 | Isoprenaline_13C-Glucose |
| Showing results 1 to 8 of 8 |
Collection:
| Collection ID: | CO001617 |
| 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: | TR001637 |
| 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) and 11 mM U-13C6-D-Glucose labelled medium (to give a final ratio of 50:50 of U-13C and U-12C D-glucose). |
Sample Preparation:
| Sampleprep ID: | SP001630 |
| 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: | CH001888 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | ZIC-pHILIC (150 x 4.6mm,5um) |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH001889 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | ZIC-pHILIC (150 x 4.6mm,5um) |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN002578 |
| Analysis Type: | MS |
| Chromatography ID: | CH001888 |
| Num Factors: | 2 |
| Num Metabolites: | 339 |
| Rt Units: | Minutes |
| Units: | Intensity |
| Analysis ID: | AN002579 |
| Analysis Type: | MS |
| Chromatography ID: | CH001889 |
| Num Factors: | 2 |
| Num Metabolites: | 341 |
| Rt Units: | Minutes |
| Units: | Intensity |
