Summary of Study ST002280
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 PR001460. The data can be accessed directly via it's Project DOI: 10.21228/M80T4P 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.
| Study ID | ST002280 |
| Study Title | Oxidative phosphorylation selectively orchestrates tissue macrophage homeostasis |
| Study Type | Observational study |
| Study Summary | In vitro studies associated oxidative phosphorylation (OXPHOS) with anti-inflammatory macrophages, while pro-inflammatory macrophages rely on glycolysis. However, the metabolic needs of macrophages in tissues (TMFs) to fulfil their homeostatic activities are incompletely understood. Here, we identified OXPHOS as highly discriminating process among TMFs from different tissues in homeostasis by analysis of RNAseq data, in both human and mouse. Impairing OXPHOS in TMFs via Tfam deletion differentially affected TMF populations. Tfam deletion resulted in reduction of alveolar macrophages (AMs) due to impaired lipid-handling capacity, leading to increased cholesterol content and cellular stress, causing cell cycle arrest in vivo. In obesity, Tfam depletion selectively ablated pro-inflammatory lipid-handling white adipose tissue macrophages (WAT-MFs), preventing insulin resistance and hepatosteatosis. Thus, OXPHOS, rather than glycolysis, distinguishes TMF populations and is critical for the maintenance of TMFs with a high lipid-handling activity, including pro-inflammatory WAT-MFs. This could provide a selective therapeutic targeting tool. |
| Institute | Spanish National Center for Cardiovascular Research (CNIC) |
| Department | Novel mechanisms of atherosclerosis |
| Laboratory | Immunobiology |
| Last Name | Mastrangelo |
| First Name | Annalaura |
| Address | Calle de Melchor Fernández Almagro, 3, Centro Nacional de Investigaciones Cardiovasculares |
| annalaura.mastrangelo@cnic.es | |
| Phone | (+34) 914531200 |
| Submit Date | 2022-09-01 |
| Num Groups | 2 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | GC-MS |
| Release Date | 2022-09-22 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001460 |
| Project DOI: | doi: 10.21228/M80T4P |
| Project Title: | Oxidative phosphorylation selectively orchestrates tissue macrophage homeostasis |
| Project Summary: | In vitro studies associated oxidative phosphorylation (OXPHOS) with anti-inflammatory macrophages, while pro-inflammatory macrophages rely on glycolysis. However, the metabolic needs of macrophages in tissues (TMFs) to fulfil their homeostatic activities are incompletely understood. Here, we identified OXPHOS as highly discriminating process among TMFs from different tissues in homeostasis by analysis of RNAseq data, in both human and mouse. Impairing OXPHOS in TMFs via Tfam deletion differentially affected TMF populations. Tfam deletion resulted in reduction of alveolar macrophages (AMs) due to impaired lipid-handling capacity, leading to increased cholesterol content and cellular stress, causing cell cycle arrest in vivo. In obesity, Tfam depletion selectively ablated pro-inflammatory lipid-handling white adipose tissue macrophages (WAT-MFs), preventing insulin resistance and hepatosteatosis. Thus, OXPHOS, rather than glycolysis, distinguishes TMF populations and is critical for the maintenance of TMFs with a high lipid-handling activity, including pro-inflammatory WAT-MFs. This could provide a selective therapeutic targeting tool. |
| Institute: | Spanish National Center for Cardiovascular Research (CNIC) |
| Department: | Novel mechanisms of atherosclerosis |
| Laboratory: | Immunobiology |
| Last Name: | Mastrangelo |
| First Name: | Annalaura |
| Address: | Calle de Melchor Fernández Almagro, 3, Centro Nacional de Investigaciones Cardiovasculares |
| Email: | annalaura.mastrangelo@cnic.es |
| Phone: | (+34) 914531200 |
Subject:
| Subject ID: | SU002366 |
| 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 | Genoptype |
|---|---|---|
| SA218456 | KO1 | Mutant |
| SA218457 | KO2R | Mutant |
| SA218458 | KO3R | Mutant |
| SA218459 | KO1R | Mutant |
| SA218460 | KO3 | Mutant |
| SA218461 | WT3R | Wild_type |
| SA218462 | WT3 | Wild_type |
| SA218463 | WT1 | Wild_type |
| SA218464 | WT1R | Wild_type |
| SA218465 | WT2 | Wild_type |
| SA218466 | WT2R | Wild_type |
| Showing results 1 to 11 of 11 |
Collection:
| Collection ID: | CO002359 |
| Collection Summary: | Mouse colonies were bred at the CNIC under specific pathogen-free conditions and on C57BL/6 background. Tfamf/f (Larsson et al., 1998) mice were kindly provided by Nils-Göran Larsson (Max Planck Institute for Biology of Ageing, Cologne, Germany). All floxed mouse lines were crossed with CD11cCre mice (Caton et al., 2007). Mice were group-housed, have not been used in previous procedures and were fed standard chow. Littermates of the same sex were randomly assigned to experimental groups. Male and female mice were used for all experiments. Mice with 6–10-weeks (adult) were used for the experiment. Bronchoalveolar lavage (BAL) was performed by inserting a venal catheter (BD) into the trachea and 3-10 washes with 0.3-1 ml FACS buffer to harvest BAL cells. |
| Collection Protocol Filename: | Protocol_AM.pdf |
| Sample Type: | Bronchoalveolar lavage |
| Collection Method: | Bronchoalveolar lavage (BAL) was performed by inserting a venal catheter (BD) into the trachea and 3-10 washes with 0.3-1 ml FACS buffer to harvest BAL cells. |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002378 |
| Treatment Summary: | Mice were group-housed and were fed standard chow. |
Sample Preparation:
| Sampleprep ID: | SP002372 |
| Sampleprep Summary: | One million CD45+ F4/80+ CD11c+ FACS-sorted alveolar macrophages (AMs) from the bronchoalveolar lavage (BAL) of adult Tfamf/f and CD11c∆Tfam mice were collected. Each sample was generated by merging FACS-sorted AMs from 13 to 30 mice. Quality Control (QC) samples (n=4) were prepared by pooling equal volumes of cell extracts from each sample by following the same protocols used for the subject samples. Samples were subjected to two freeze–thaw cycles for metabolism quenching and complete metabolite extraction, specifically by placing the samples at -80ºC for 15 min and thawing them on ice for 10 min with brief vortex-mixing. The samples were then centrifuged at 20,000 xg at 4°C for 10 min and the supernatant collected. The supernatant was evaporated to dryness (SpeedVac Concentrator, Thermo Fisher Scientific, Waltham, MA, USA) and derivatized with 10 μl O-methoxyamine hydrochloride (15mg/mL) in pyridine and 10 μl N,O-bis(trimethylsilyl)trifluoroacetamide in 1% trimethylchlorosilane. Finally, 100 μl of heptane containing 10 ppm of 4-nitrobenzoic acid (IS) was used as internal standard to monitor sample injection |
| Sampleprep Protocol Filename: | Protocol_AM.pdf |
Combined analysis:
| Analysis ID | AN003724 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890B |
| Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| MS Type | EI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 7250 |
| Ion Mode | POSITIVE |
| Units | relative abundance |
Chromatography:
| Chromatography ID: | CH002758 |
| Chromatography Summary: | Derivative samples (2 μL) were injected into a GC column DB5–MS (30 m length, 0.250 mm i.d., 0.25 μm film 95% dimethyl/5% diphenylpolysiloxane) with a pre–column (10 m J&W integrated with Agilent 122–5532G). The temperature gradient was programmed at 60 °C (held for 1 min), with a ramping increase rate of 10 °C/min up to 325°C (held for 10 min). The total analysis time was 37.5 min. Two analytical replicates for each sample were injected. |
| Methods Filename: | Protocol_AM.pdf |
| Instrument Name: | Agilent 7890B |
| Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| Chromatography Type: | GC |
MS:
| MS ID: | MS003472 |
| Analysis ID: | AN003724 |
| Instrument Name: | Agilent 7250 |
| Instrument Type: | QTOF |
| MS Type: | EI |
| MS Comments: | The EI source was operated at 70 eV whereas the mass spectrometer operated in the scan mode over a mass range of m/z 50–600. Metabolite deconvolution and identification were carried out using Agilent MassHunter Unknowns Analysis version B.07.00, then, data was aligned in Agilent Mass Profiler Professional version B.12.1 and exported to Agilent MassHunter Quantitative Analysis version B.07.00. Metabolites were identified by comparing their retention time, retention index and mass fragmentation patterns with those available in an in-house library including both the NIST mass spectral database (version 2017) and Fiehn RTL library (version 2008). The different derivatives that were generated from the silylated compounds were unified by summing the abundance of all derivatives from the same metabolite. Finally, the median relative area of the two analytical replicates of the same sample was computed and used for subsequent statistical analysis. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | Protocol_AM.pdf |
