Summary of Study ST004113

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.

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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 IDST004113
Study TitleMetabolic changes in NLRP3 knockout and VRAC knockdown THP-1 derived macrophages following NLRP3 inflammasome activation
Study TypeExploratory MS
Study SummaryUsing 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). Inhibiting VRAC, or supplementation of taurine, restores the ionic balance, abrogates IL-1beta release and reduces cellular cytotoxicity in THP-1 derived macrophages.
Institute
Imperial College London
Last NameRossi-Smith
First NamePeter
AddressHammersmith Campus, London, London, W12 0NN, United Kingdom
Emailp.rossi@imperial.ac.uk
Phone07860694004
Submit Date2025-08-13
Raw Data AvailableYes
Raw Data File Type(s)mzML, d
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:SU004262
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Genotype Strain:THP-1 cell line

Factors:

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

mb_sample_id local_sample_id Treatment Sample source
SA476121THP-1_31Control NLRP3 KO
SA476122THP-1_35Control NLRP3 KO
SA476123THP-1_34Control NLRP3 KO
SA476124THP-1_33Control NLRP3 KO
SA476125THP-1_32Control NLRP3 KO
SA476126THP-1_50Control VRAC KD
SA476127THP-1_49Control VRAC KD
SA476128THP-1_48Control VRAC KD
SA476129THP-1_47Control VRAC KD
SA476130THP-1_46Control VRAC KD
SA476131THP-1_2Control WT
SA476132THP-1_1Control WT
SA476133THP-1_5Control WT
SA476134THP-1_4Control WT
SA476135THP-1_3Control WT
SA476136THP-1_39LPS+ATP NLRP3 KO
SA476137THP-1_37LPS+ATP NLRP3 KO
SA476138THP-1_40LPS+ATP NLRP3 KO
SA476139THP-1_38LPS+ATP NLRP3 KO
SA476140THP-1_36LPS+ATP NLRP3 KO
SA476141THP-1_52LPS+ATP VRAC KD
SA476142THP-1_51LPS+ATP VRAC KD
SA476143THP-1_53LPS+ATP VRAC KD
SA476144THP-1_54LPS+ATP VRAC KD
SA476145THP-1_55LPS+ATP VRAC KD
SA476146THP-1_9LPS+ATP WT
SA476147THP-1_6LPS+ATP WT
SA476148THP-1_8LPS+ATP WT
SA476149THP-1_10LPS+ATP WT
SA476150THP-1_7LPS+ATP WT
SA476151THP-1_45LPS+NG NLRP3 KO
SA476152THP-1_44LPS+NG NLRP3 KO
SA476153THP-1_43LPS+NG NLRP3 KO
SA476154THP-1_42LPS+NG NLRP3 KO
SA476155THP-1_41LPS+NG NLRP3 KO
SA476156THP-1_56LPS+NG VRAC KD
SA476157THP-1_57LPS+NG VRAC KD
SA476158THP-1_58LPS+NG VRAC KD
SA476159THP-1_59LPS+NG VRAC KD
SA476160THP-1_60LPS+NG VRAC KD
SA476161THP-1_15LPS+NG WT
SA476162THP-1_11LPS+NG WT
SA476163THP-1_12LPS+NG WT
SA476164THP-1_13LPS+NG WT
SA476165THP-1_14LPS+NG WT
Showing results 1 to 45 of 45

Collection:

Collection ID:CO004255
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:TR004271
Treatment Summary:All inflammasome activation reagents were sourced from InvivoGen, unless otherwise specified. NLRP3 inflammasome activation was induced in THP-1 derived macrophages 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 µM nigericin (N7143, Sigma-Aldrich) for ~45 minutes

Sample Preparation:

Sampleprep ID:SP004268
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:CH005181
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:AN006820
Analysis Type:MS
Chromatography ID:CH005181
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST004113_AN006820_Results.txt
Units:Area
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