Summary of Study ST004106
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 PR002581. The data can be accessed directly via it's Project DOI: 10.21228/M81C34 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 | ST004106 |
| Study Title | Mitochondrial complex III-derived reactive oxygen species amplify immunometabolic changes in astrocytes and promote dementia pathology |
| Study Type | Targeted metabolomics |
| Study Summary | Neurodegenerative disorders alter mitochondrial functions, including the production of reactive oxygen species (ROS). Mitochondrial complex III (CIII) generates ROS implicated in redox signaling, but its triggers, temporal dynamics, targets, and disease relevance are not clear. Using site-selective suppressors and genetic manipulations together with live mitochondrial ROS imaging and multiomic profiling, we found that CIII is a dominant source of ROS production in astrocytes exposed to neuropathology-related stimuli. Astrocytic CIII-ROS production was dependent on nuclear factor-κB (NF-κB) and the mitochondrial sodium-calcium exchanger (NCLX) and caused oxidation of select cysteines within immune- and metabolism-associated proteins linked to neurological disease. CIII-ROS amplified metabolomic and pathology-associated transcriptional changes in astrocytes, with STAT3 activity as a major mediator, and facilitated neuronal toxicity. Therapeutic suppression of CIII-ROS in mice decreased dementia-linked tauopathy and neuroimmune cascades and extended lifespan. Our findings establish CIII-ROS as an important immunometabolic signal transducer and tractable therapeutic target in neurodegenerative disease. For the metabolomics data, we treated astrocytes with interleukin-1alpha (IL-1ɑ) to drive immune signaling and co-treated cells with TPCA-1, an inhibitor of NF-κB, or S3QEL1.2, a suppressor of ROS production specifically from mitochondrial complex III. These inhibitor co-treatments were designed to determine if IL-1ɑ-mediated metabolomic changes were driven by NF-κB activation and/or complex III ROS production. |
| Institute | Weill Cornell Medicine |
| Department | Appel Alzheimer's Disease Research Institute, Feil Family Brain and Mind Research Institute |
| Laboratory | Orr Laboratory |
| Last Name | Barnett |
| First Name | Daniel |
| Address | 413 East 69th St, New York, NY, 10021, USA |
| dmb4001@med.cornell.edu | |
| Phone | 5854659449 |
| Submit Date | 2025-08-04 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-09-07 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002581 |
| Project DOI: | doi: 10.21228/M81C34 |
| Project Title: | Mitochondrial complex III-derived reactive oxygen species amplify immunometabolic changes in astrocytes and promote dementia pathology |
| Project Summary: | Neurodegenerative disorders alter mitochondrial functions, including the production of reactive oxygen species (ROS). Mitochondrial complex III (CIII) generates ROS implicated in redox signaling, but its triggers, temporal dynamics, targets, and disease relevance are not clear. Using site-selective suppressors and genetic manipulations together with live mitochondrial ROS imaging and multiomic profiling, we found that CIII is a dominant source of ROS production in astrocytes exposed to neuropathology-related stimuli. Astrocytic CIII-ROS production was dependent on nuclear factor-κB (NF-κB) and the mitochondrial sodium-calcium exchanger (NCLX) and caused oxidation of select cysteines within immune- and metabolism-associated proteins linked to neurological disease. CIII-ROS amplified metabolomic and pathology-associated transcriptional changes in astrocytes, with STAT3 activity as a major mediator, and facilitated neuronal toxicity. Therapeutic suppression of CIII-ROS in mice decreased dementia-linked tauopathy and neuroimmune cascades and extended lifespan. Our findings establish CIII-ROS as an important immunometabolic signal transducer and tractable therapeutic target in neurodegenerative disease. |
| Institute: | Weill Cornell Medicine |
| Department: | Appel Alzheimer's Disease Research Institute, Feil Family Brain and Mind Research Institute |
| Laboratory: | Orr Laboratory |
| Last Name: | Barnett |
| First Name: | Daniel |
| Address: | 413 East 69th St, New York, NY, 10021, USA |
| Email: | dmb4001@med.cornell.edu |
| Phone: | 5854659449 |
| Funding Source: | National Institute on Aging, National Institutes of Health |
Subject:
| Subject ID: | SU004253 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | Wild-type |
| Age Or Age Range: | Postnatal day 2-3 |
| Cell Primary Immortalized: | Primary astrocyte derived from cortex and hippocampus |
Factors:
Subject type: Cultured cells; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Treatment |
|---|---|---|---|
| SA474902 | IL1a_S4 | Cortex/Hippocampus | IL-1a |
| SA474903 | IL1a_S2 | Cortex/Hippocampus | IL-1a |
| SA474904 | IL1a_S1 | Cortex/Hippocampus | IL-1a |
| SA474905 | IL1a_S3 | Cortex/Hippocampus | IL-1a |
| SA474906 | IL1aS_S1 | Cortex/Hippocampus | IL-1a+S3QEL1.2 |
| SA474907 | IL1aS_S4 | Cortex/Hippocampus | IL-1a+S3QEL1.2 |
| SA474908 | IL1aS_S2 | Cortex/Hippocampus | IL-1a+S3QEL1.2 |
| SA474909 | IL1aS_S3 | Cortex/Hippocampus | IL-1a+S3QEL1.2 |
| SA474910 | IL1aT_S1 | Cortex/Hippocampus | IL-1a+TPCA-1 |
| SA474911 | IL1aT_S2 | Cortex/Hippocampus | IL-1a+TPCA-1 |
| SA474912 | IL1aT_S3 | Cortex/Hippocampus | IL-1a+TPCA-1 |
| SA474913 | IL1aT_S4 | Cortex/Hippocampus | IL-1a+TPCA-1 |
| SA474914 | Veh_S1 | Cortex/Hippocampus | Vehicle |
| SA474915 | Veh_S4 | Cortex/Hippocampus | Vehicle |
| SA474916 | Veh_S2 | Cortex/Hippocampus | Vehicle |
| SA474917 | Veh_S3 | Cortex/Hippocampus | Vehicle |
| Showing results 1 to 16 of 16 |
Collection:
| Collection ID: | CO004246 |
| Collection Summary: | Primary astrocytes were isolated from the cortex and hippocampus of postnatal wild-type mice. All cultures were maintained at 37°C in a humidified 5% CO2-containing atmosphere. Astrocytes were grown, maintained, and treated in high-glucose DMEM without glutamine or pyruvate (Corning) and supplemented with 20% heat-inactivated FBS (VWR), 1X GlutaMAX (ThermoFisher), and 1 mM sodium pyruvate (ThermoFisher). Cells were cultured in 10cm plates that were pre-coated with sterile-filtered 100 µg/mL poly-D-lysine. |
| Sample Type: | Astrocyte cells |
Treatment:
| Treatment ID: | TR004262 |
| Treatment Summary: | Astrocytes were first pre-treated for 1 hour with Vehicle (DMSO) or 1 µM TPCA-1 (Cayman Chemicals), then co-treated for 6 hours with Vehicle, 3 ng/mL IL-1α (Sigma), IL-1α + 3 µM S3QEL1.2 (WuXi Apptec), or IL-1α + 1 µM TPCA-1 with an n = 4 replicates per condition. |
Sample Preparation:
| Sampleprep ID: | SP004259 |
| Sampleprep Summary: | Following treatment, polar metabolites were extracted on dry ice using pre-chilled 80% methanol (-80 °C). The extract was dried with a Speedvac, and redissolved in HPLC grade water before it was applied to the hydrophilic interaction chromatography LC-MS. |
Chromatography:
| Chromatography ID: | CH005171 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | SeQuant ZIC- pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | NA |
| Flow Gradient: | The chromatographic gradient ran from 85% to 30% A in 20 min followed by a wash with 30% A and re-equilibration at 85% A |
| Flow Rate: | 150 μL/min |
| Solvent A: | 100% acetonitrile |
| Solvent B: | 100% water; 0.1% ammonium hydroxide; 20 mM ammonium acetate |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN006808 |
| Analysis Type: | MS |
| Analysis Protocol File: | polar_metabolomics_method.pdf |
| Chromatography ID: | CH005171 |
| Num Factors: | 4 |
| Num Metabolites: | 203 |
| Units: | Signal intensity (protein normalized) |
