Summary of Study ST004115
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 PR002586. The data can be accessed directly via it's Project DOI: 10.21228/M8CN8W 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 | ST004115 |
| Study Title | Metabolic changes in murine macrophages following NLRP3 inflammasome activation |
| Study Type | Exploratory MS |
| Study Summary | The intersection of immunology and metabolism, known as immunometabolism, explores the interactions between immune responses and metabolic changes. Inflammasomes form an integral part of the innate immune system and are equipped with NLR or ALR receptors capable of detecting a wide array of stimuli triggered by infections or cellular damage. Upon activation, these inflammasomes are involved in the release of inflammatory cytokines and can trigger a regulated type of cell death known as pyroptosis. Like other immune responses, inflammasome activation also induces changes in metabolic pathways such as the tricarboxylic acid (TCA) cycle. However, the role of other metabolic pathways in response to activation of inflammasomes remains less explored. Here, by employing a metabolomic approach on murine macrophages, we found that activation of inflammasomes (NLRP3, AIM2 or NLRC4) induced metabolic shifts not only within the TCA cycle, but also extends its impact to Sulphur metabolism. Furthermore, through a rigorous cross-species analysis that compared human and mouse responses (shown in Taurine transport is a critical modulator of ionic fluxes during NLRP3 inflammasome activation project), we uncovered a notable downregulation of taurine metabolism following NLRP3 activation. This intriguing discovery highlighted a conserved regulatory mechanism and identified intracellular depletion of taurine and hypotaurine as a putative checkpoint in NLRP3 activation pathway. |
| Institute | Imperial College London |
| Last Name | Rossi-Smith |
| First Name | Peter |
| Address | Hammersmith Campus, London, London, W12 0NN, United Kingdom |
| p.rossi@imperial.ac.uk | |
| Phone | 07860694004 |
| Submit Date | 2025-08-13 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, d |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-08-17 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002586 |
| Project DOI: | doi: 10.21228/M8CN8W |
| Project Title: | Taurine transport is a critical modulator of ionic fluxes during NLRP3 inflammasome activation |
| Project Type: | MS exploratory analysis |
| Project Summary: | Metabolic regulation is a key feature of inflammasome activation and effector function. Using metabolomic approaches, we show that downregulation of taurine metabolism is crucial for NLRP3 inflammasome activation. Following NLRP3 activation stimuli, taurine rapidly egresses to the extracellular compartment. Taurine efflux is facilitated primarily by the volume-regulated anion channel (VRAC). Loss of intracellular taurine impairs sodium-potassium ATPase pump activity, promoting ionic dysregulation and disrupting ionic fluxes. Inhibiting VRAC, or supplementation of taurine, restores the ionic balance, abrogates IL-1beta release and reduces cellular cytotoxicity in macrophages. We further demonstrate that the protective effect of taurine is diminished when sodium-potassium ATPase is inhibited, highlighting the pump’s role in taurine-mediated protection. Finally, taurine metabolism is significantly associated with the development of tuberculosis-associated immune reconstitution inflammatory syndrome, a systemic hyperinflammatory condition known to be mediated by inflammasome activation. Altogether, we identified a critical metabolic pathway that modulates inflammasome activation and drives disease pathogenesis. |
| Institute: | Imperial College London |
| Department: | Department of Infectious Disease |
| Laboratory: | Lai's Lab |
| Last Name: | Rossi-Smith |
| First Name: | Peter |
| Address: | Hammersmith Campus, London, London, W12 0NN, United Kingdom |
| Email: | p.rossi@imperial.ac.uk |
| Phone: | 07860694004 |
| Funding Source: | This work was supported by an MRC CDA fellowship (MR/R008922/1) to R.P.J.L. and in part by the NIHR Imperial Biomedical Research Centre and an NIH R01 grant (5R01AI145436) to R.J.W. and R.P.J.L. D.C.T. is supported by a Wellcome-Beit Prize Trust Clinical Research Career Development Fellowship and the Burman Fund from Imperial College London. J.P.G. is supported by MRC research grant (MR/W028867/1). A.E.D. is supported by an MRC CDA fellowship (MR/V009591/1). R.J.W., M.S.S. and J.I.M. are supported by The Francis Crick Institute, which receives its core funding from Cancer Research UK (CC2206), the UK Medical Research Council (CC2206), and the Wellcome Trust (CC2206). T.E. and C.W. acknowledge funding from the BBSRC grant (BB/W002345/1). T.E. acknowledges partial support from UKRI BBSRC grant BB/T007974/1, European Union projects HUMAN (EC101073062) and BiACEM (EC101079370). G.M. was supported by the Wellcome Trust (098316, 214321/Z/18/Z, and 203135/Z/16/Z) and the South African Research Chairs Initiative of the Department of Science and Technology and National Research Foundation (NRF) of South Africa (Grant no. 64787). The funders had no role in the study design, data collection, data analysis, data interpretation, or writing of this report. The opinions, findings and conclusions expressed in this manuscript reflect those of the authors alone. This research was funded, in part, by the Wellcome Trust. For the purpose of open access, the authors have applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. |
| Contributors: | Dr. Rachel Lai |
Subject:
| Subject ID: | SU004264 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6 |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment | Sample source |
|---|---|---|---|
| SA476204 | WT2_8 | ATP | Macrophages |
| SA476205 | WT2_10 | ATP | Macrophages |
| SA476206 | WT2_9 | ATP | Macrophages |
| SA476207 | WT2_6 | ATP | Macrophages |
| SA476208 | WT2_7 | ATP | Macrophages |
| SA476209 | WT2_2 | Control | Macrophages |
| SA476210 | WT2_1 | Control | Macrophages |
| SA476211 | WT2_5 | Control | Macrophages |
| SA476212 | WT2_3 | Control | Macrophages |
| SA476213 | WT2_4 | Control | Macrophages |
| SA476214 | WT2_22 | LPS+ATP | Macrophages |
| SA476215 | WT2_25 | LPS+ATP | Macrophages |
| SA476216 | WT2_24 | LPS+ATP | Macrophages |
| SA476217 | WT2_23 | LPS+ATP | Macrophages |
| SA476218 | WT2_21 | LPS+ATP | Macrophages |
| SA476219 | WT2_30 | LPS+NG | Macrophages |
| SA476220 | WT2_29 | LPS+NG | Macrophages |
| SA476221 | WT2_28 | LPS+NG | Macrophages |
| SA476222 | WT2_27 | LPS+NG | Macrophages |
| SA476223 | WT2_26 | LPS+NG | Macrophages |
| SA476224 | WT2_20 | LPS | Macrophages |
| SA476225 | WT2_19 | LPS | Macrophages |
| SA476226 | WT2_18 | LPS | Macrophages |
| SA476227 | WT2_17 | LPS | Macrophages |
| SA476228 | WT2_16 | LPS | Macrophages |
| SA476229 | WT2_15 | NG | Macrophages |
| SA476230 | WT2_14 | NG | Macrophages |
| SA476231 | WT2_13 | NG | Macrophages |
| SA476232 | WT2_12 | NG | Macrophages |
| SA476233 | WT2_11 | NG | Macrophages |
| Showing results 1 to 30 of 30 |
Collection:
| Collection ID: | CO004257 |
| Collection Summary: | Following in vitro experiments, mBMDM metabolites were quenched by washing the cells twice with ice-cold AUTOMacs Rinsing Solution (Miltenyi Biotec), before a methanol (10767665, Fisher Chemical):water (10505904, Fisher Chemical) (4:1 v/v) solution was added and macrophages were gently scrapped. Lysed macrophages were re-suspended in chloroform (10615492, Fisher Chemical) and submitted to 3 cycles: vortex for 0.5 min and placed on ice for 5 min. Following the last vortexing cycle the samples were stored at -80°C for no less than 12 hours. |
| Collection Protocol Filename: | LC-MS_protocol.pdf |
| Sample Type: | Macrophages |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR004273 |
| Treatment Summary: | All inflammasome activation reagents were sourced from InvivoGen, unless otherwise specified. NLRP3 inflammasome activation was induced in mBMDM by priming with 500 ng/mL LPS (tlrl-peklps) for 3.5 hours, followed by stimulation with either 5 mM ATP (tlrl-atpl) or 20 uM nigericin (N7143, Sigma-Aldrich) for ~45 minutes. |
Sample Preparation:
| Sampleprep ID: | SP004270 |
| Sampleprep Summary: | After overnight incubation in -80°C, water was added to generate a biphasic solution with a final dilution of 3:2:4 (v/v) chloroform:water:methanol. The samples were then vortexed and centrifuged at 14,000 rpm for 10 min. at 0°C. The top layer containing polar metabolites (avoiding the interface) was concentrated using a SpeedVac. The dried samples were resuspended in 75 µL of 30% methanol and 2% acetonitrile (10001334, Fisher Chemical) and stored at -80°C until metabolomics analyses were carried out. |
| Extract Storage: | -80℃ |
Chromatography:
| Chromatography ID: | CH005183 |
| Chromatography Summary: | Samples were analyzed using an Agilent 1290 Infinity II UHPLC coupled with Agilent 6546 LC/QTOF. The system was equipped with an Agilent Poroshell 120 HILIC-Z column (2.1 x 150 mm, 2.1 µm). A 2 µL sample volume was injected, and the chromatographic separation was performed at 15°C with a flow rate of 400 µL/min using an elution gradient. Mobile phases A (20 mM ammonium acetate, 5 µM medronic acid, pH 9.3) and B (acetonitrile) were used with the following gradient: 0-1 min, 85% B; 1-8 min, 75% B; 8-12 min, 60% B; 12-19.10 min, 10% B; 19.10-24 min, 85% B. |
| Instrument Name: | Agilent 1290 Infinity |
| Column Name: | Agilent InfinityLab Poroshell 120 EC-C8 (150 x 2.1 mm, 2.7 µm) |
| Column Temperature: | 15°C |
| Flow Gradient: | 0-1 min, 85% B; 1-8 min, 75% B; 8-12 min, 60% B; 12-19.10 min, 10% B; 19.10-24 min, 85% B |
| Flow Rate: | 400 µL/min |
| Solvent A: | 100% Water; 20 mM Ammonium acetate; 5 µM Medronic acid (pH 9.3) |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN006822 |
| Analysis Type: | MS |
| Chromatography ID: | CH005183 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004115_AN006822_Results.txt |
| Units: | Area |
| Analysis ID: | AN006823 |
| Analysis Type: | MS |
| Chromatography ID: | CH005183 |
| Has Mz: | 1 |
| Has Rt: | 1 |
| Rt Units: | Minutes |
| Results File: | ST004115_AN006823_Results.txt |
| Units: | Area |
