Summary of Study ST004111

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.

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Study IDST004111
Study TitleMetabolic changes in human macrophages following NLRP3 activation
Study TypeExploratory MS
Study SummaryThe 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 human macrophages, we found that activation of inflammasomes NLRP3 induced metabolic shifts not only within the TCA cycle, but also extends its impact to Sulphur metabolism, more specifically downregulation of the taurine metabolism.
Institute
Imperial College London
Last NameRossi-Smith
First NamePeter
AddressHammersmith Campus, London, London, W12 0NN, United Kingdom
Emailp.rossi@imperial.ac.uk
Phone07860694004
Submit Date2025-08-11
Raw Data AvailableYes
Raw Data File Type(s)mzML, raw(Thermo)
Analysis Type DetailLC-MS
Release Date2025-08-15
Release Version1
Peter Rossi-Smith Peter Rossi-Smith
https://dx.doi.org/10.21228/M8CN8W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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:SU004263
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

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

mb_sample_id local_sample_id Treatment Sample source
SA476166220119_PR211125_h06ATP Macrophages
SA476167220119_PR211125_h02ATP Macrophages
SA476168220119_PR211125_h01ATP Macrophages
SA476169220119_PR211125_h04ATP Macrophages
SA476170220119_PR211125_h05ATP Macrophages
SA476171220119_PR211125_h03ATP Macrophages
SA476172220119_PR211125_h36Control Macrophages
SA476173220119_PR211125_h35Control Macrophages
SA476174220119_PR211125_h34Control Macrophages
SA476175220119_PR211125_h33Control Macrophages
SA476176220119_PR211125_h32Control Macrophages
SA476177220119_PR211125_h31Control Macrophages
SA476178220119_PR211125_h19LPS+ATP Macrophages
SA476179220119_PR211125_h24LPS+ATP Macrophages
SA476180220119_PR211125_h23LPS+ATP Macrophages
SA476181220119_PR211125_h21LPS+ATP Macrophages
SA476182220119_PR211125_h22LPS+ATP Macrophages
SA476183220119_PR211125_h20LPS+ATP Macrophages
SA476184220119_PR211125_h26LPS+NG Macrophages
SA476185220119_PR211125_h30LPS+NG Macrophages
SA476186220119_PR211125_h28LPS+NG Macrophages
SA476187220119_PR211125_h27LPS+NG Macrophages
SA476188220119_PR211125_h29LPS+NG Macrophages
SA476189220119_PR211125_h25LPS+NG Macrophages
SA476190220119_PR211125_h13LPS Macrophages
SA476191220119_PR211125_h14LPS Macrophages
SA476192220119_PR211125_h15LPS Macrophages
SA476193220119_PR211125_h17LPS Macrophages
SA476194220119_PR211125_h16LPS Macrophages
SA476195220119_PR211125_h18LPS Macrophages
SA476196220119_PR211125_PBQC_h6NEGMSMS_PBQC_Neg Macrophages
SA476197220119_PR211125_PBQC_h5POSMSMS_PBQC_Pos Macrophages
SA476198220119_PR211125_h12NG Macrophages
SA476199220119_PR211125_h11NG Macrophages
SA476200220119_PR211125_h10NG Macrophages
SA476201220119_PR211125_h09NG Macrophages
SA476202220119_PR211125_h08NG Macrophages
SA476203220119_PR211125_h07NG Macrophages
Showing results 1 to 38 of 38

Collection:

Collection ID:CO004256
Collection Summary:Following in vitro experiments, hBMM 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

Treatment:

Treatment ID:TR004272
Treatment Summary:All inflammasome activation reagents were sourced from InvivoGen, unless otherwise specified. NLRP3 inflammasome activation was induced in hBMM 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:SP004269
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 uL of 30% methanol and 2% acetonitrile (10001334, Fisher Chemical) and stored at -80°C until metabolomics analyses were carried out.

Chromatography:

Chromatography ID:CH005182
Chromatography Summary:Samples were analyzed using a Dionex UltiMate Liquid Chromatography (LC) system (Thermo Scientific) with a ZIC-pHILIC (150 mm x 4.6 mm, 5 µm particle) column (Merck Sequant). The mobile phase contained 20 mM ammonium carbonate in water – solvent A (Optima HPLC grade, Sigma Aldrich), and acetonitrile – solvent B (Optima HPLC grade, Sigma Aldrich). Samples were separated using an elution gradient: 15 min of 4:1 (Solvent A:Solvent B), 5 min 95:5 (Solvent A:Solvent B) and 5 min re-equilibration. The samples were kept at 4°C during the whole run, the mobile phase had a flow rate of 300 uL/min and the column temperature was 25°C
Instrument Name:Agilent 1290 Infinity
Column Name:Merck SeQuant ZIC-pHILIC (150 x 4.6 mm, 5 µm)
Column Temperature:25°C
Flow Gradient:15 min of 4:1 (Solvent A:Solvent B), 5 min 95:5 (Solvent A:Solvent B) and 5 min re-equilibration.
Flow Rate:300 µL/min
Solvent A:100% Water; 20 mM Ammonium carbonate
Solvent B:100% Acetonitrile
Chromatography Type:HILIC

Analysis:

Analysis ID:AN006821
Analysis Type:MS
Chromatography ID:CH005182
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST004111_AN006821_Results.txt
Units:Area
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