Summary of Study ST004164

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 PR002624. The data can be accessed directly via it's Project DOI: 10.21228/M8G84H 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 IDST004164
Study TitleEquilibrative Nucleoside Transporter 2 Modulates Inosine Catabolism to Influence Astrocyte Metabolism and Reactivity
Study SummaryIn this study, we demonstrate that inosine functions as an essential energy substrate for astrocytes under inflammatory conditions. Targeted metabolomic datasets were generated from mouse primary astrocytes after 24 hours of cytokine cocktail treatment. Metabolomic profiling, combined with pharmacological approaches, revealed that Ent2 regulates inosine efflux and is critical for maintaining astrocytic energy balance. Pharmacological inhibition of inosine utilization in energy-producing pathways with Forodesine led to disrupted energy homeostasis and increased astrocytic reactivity. The datasets include both raw and processed metabolomic files. Metabolomic analyses were performed at the Metabolomics Core Facility, Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.
Institute
Institute of Biomedical Sciences, Academia Sinica
Last NameChern
First NameYijuang
Address128 Section 2, Academia Road, Nankang, Taipei, 115201, Taiwan
Emailbmychern@ibms.sinica.edu.tw
Phone+886-2-26523913
Submit Date2025-08-28
Raw Data AvailableYes
Raw Data File Type(s)mzML
Analysis Type DetailLC-MS
Release Date2025-09-30
Release Version1
Yijuang Chern Yijuang Chern
https://dx.doi.org/10.21228/M8G84H
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002624
Project DOI:doi: 10.21228/M8G84H
Project Title:Equilibrative Nucleoside Transporter 2 Modulates Inosine Catabolism to Influence Astrocyte Metabolism and Reactivity
Project Summary:Astrocytes are central regulators of brain function and exhibit diverse reactive states in response to injury and inflammation. Although the mechanisms underlying these states remain debated, cellular energy metabolism is thought to play a crucial role. This project aimed to define the role of equilibrative nucleoside transporter 2 (Ent2) in regulating astrocytic responses to inflammatory stimulation, with a particular focus on its function in inosine metabolism and energy balance. Using mouse primary astrocytes exposed to cytokine-induced activation, together with targeted metabolomic profiling and pharmacological manipulation, we examined how Ent2 activity influences purine metabolism and reactivity. Our findings show that inosine is an essential energy substrate during inflammation and that Ent2 is required to regulate inosine efflux and maintain ATP production. Loss of Ent2, or pharmacological inhibition of inosine catabolism, disrupted energy homeostasis, impaired mitochondrial function, and enhanced astrocytic reactivity. These results establish an Ent2-inosine-PNP axis as a critical pathway linking purine metabolism to astrocytic reactivity.
Institute:Institute of Biomedical Sciences, Academia Sinica
Last Name:Chern
First Name:Yijuang
Address:128 Section 2, Academia Road, Nankang, Taipei, 115201, Taiwan
Email:bmychern@ibms.sinica.edu.tw
Phone:+886-2-2652-3913

Subject:

Subject ID:SU004315
Subject Type:Cultured cells
Subject Species:Mus musculus
Taxonomy ID:10090

Factors:

Subject type: Cultured cells; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Genotype treatment
SA481171KO_3_Cytokinemouse primary astrocytes Ent10 knockout TNF𝝰/IL-1𝝰
SA481172p_KO_3_Cytokinemouse primary astrocytes Ent10 knockout TNF𝝰/IL-1𝝰
SA481173KO_4_Cytokinemouse primary astrocytes Ent11 knockout TNF𝝰/IL-1𝝰
SA481174p_KO_4_Cytokinemouse primary astrocytes Ent11 knockout TNF𝝰/IL-1𝝰
SA481175p_KO_5_Cytokinemouse primary astrocytes Ent12 knockout TNF𝝰/IL-1𝝰
SA481176KO_5_Cytokinemouse primary astrocytes Ent12 knockout TNF𝝰/IL-1𝝰
SA481177p_KO_6_Cytokinemouse primary astrocytes Ent13 knockout TNF𝝰/IL-1𝝰
SA481178KO_6_Cytokinemouse primary astrocytes Ent13 knockout TNF𝝰/IL-1𝝰
SA481179p_KO_7_Cytokinemouse primary astrocytes Ent14 knockout TNF𝝰/IL-1𝝰
SA481180KO_7_Cytokinemouse primary astrocytes Ent14 knockout TNF𝝰/IL-1𝝰
SA481181p_KO_1_CytokinePlusForomouse primary astrocytes Ent15 knockout TNF𝝰/IL-1𝝰/Forodesine
SA481182KO_1_CytokinePlusForomouse primary astrocytes Ent15 knockout TNF𝝰/IL-1𝝰/Forodesine
SA481183KO_2_CytokinePlusForomouse primary astrocytes Ent16 knockout TNF𝝰/IL-2𝝰/Forodesine
SA481184p_KO_2_CytokinePlusForomouse primary astrocytes Ent16 knockout TNF𝝰/IL-2𝝰/Forodesine
SA481185p_KO_3_CytokinePlusForomouse primary astrocytes Ent17 knockout TNF𝝰/IL-3𝝰/Forodesine
SA481186KO_3_CytokinePlusForomouse primary astrocytes Ent17 knockout TNF𝝰/IL-3𝝰/Forodesine
SA481187KO_4_CytokinePlusForomouse primary astrocytes Ent18 knockout TNF𝝰/IL-4𝝰/Forodesine
SA481188p_KO_4_CytokinePlusForomouse primary astrocytes Ent18 knockout TNF𝝰/IL-4𝝰/Forodesine
SA481189p_KO_5_CytokinePlusForomouse primary astrocytes Ent19 knockout TNF𝝰/IL-5𝝰/Forodesine
SA481190KO_5_CytokinePlusForomouse primary astrocytes Ent19 knockout TNF𝝰/IL-5𝝰/Forodesine
SA481193KO_6_CytokinePlusForomouse primary astrocytes Ent20 knockout TNF𝝰/IL-6𝝰/Forodesine
SA481194p_KO_6_CytokinePlusForomouse primary astrocytes Ent20 knockout TNF𝝰/IL-6𝝰/Forodesine
SA481195p_KO_7_CytokinePlusForomouse primary astrocytes Ent21 knockout TNF𝝰/IL-7𝝰/Forodesine
SA481196KO_7_CytokinePlusForomouse primary astrocytes Ent21 knockout TNF𝝰/IL-7𝝰/Forodesine
SA481191KO_1_Ctrlmouse primary astrocytes Ent2 knockout Vehicle
SA481192p_KO_1_Ctrlmouse primary astrocytes Ent2 knockout Vehicle
SA481197p_KO_2_Ctrlmouse primary astrocytes Ent3 knockout Vehicle
SA481198KO_2_Ctrlmouse primary astrocytes Ent3 knockout Vehicle
SA481199p_KO_3_Ctrlmouse primary astrocytes Ent4 knockout Vehicle
SA481200KO_3_Ctrlmouse primary astrocytes Ent4 knockout Vehicle
SA481201KO_4_Ctrlmouse primary astrocytes Ent5 knockout Vehicle
SA481202p_KO_4_Ctrlmouse primary astrocytes Ent5 knockout Vehicle
SA481203KO_5_Ctrlmouse primary astrocytes Ent6 knockout Vehicle
SA481204p_KO_5_Ctrlmouse primary astrocytes Ent6 knockout Vehicle
SA481205KO_6_Ctrlmouse primary astrocytes Ent7 knockout Vehicle
SA481206p_KO_6_Ctrlmouse primary astrocytes Ent7 knockout Vehicle
SA481207KO_1_Cytokinemouse primary astrocytes Ent8 knockout TNF𝝰/IL-1𝝰
SA481208p_KO_1_Cytokinemouse primary astrocytes Ent8 knockout TNF𝝰/IL-1𝝰
SA481209p_KO_2_Cytokinemouse primary astrocytes Ent9 knockout TNF𝝰/IL-1𝝰
SA481210KO_2_Cytokinemouse primary astrocytes Ent9 knockout TNF𝝰/IL-1𝝰
SA481211p_WT_3_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481212p_WT_7_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481213p_WT_6_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481214p_WT_5_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481215p_WT_4_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481216WT_2_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481217p_WT_2_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481218p_WT_1_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481219WT_1_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481220WT_3_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481221WT_4_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481222WT_7_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481223WT_6_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481224WT_5_Cytokinemouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰
SA481225WT_1_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰/Forodesine
SA481226p_WT_1_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-1𝝰/Forodesine
SA481227WT_2_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-2𝝰/Forodesine
SA481228p_WT_2_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-2𝝰/Forodesine
SA481229WT_3_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-3𝝰/Forodesine
SA481230p_WT_3_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-3𝝰/Forodesine
SA481231WT_4_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-4𝝰/Forodesine
SA481232p_WT_4_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-4𝝰/Forodesine
SA481233p_WT_5_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-5𝝰/Forodesine
SA481234WT_5_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-5𝝰/Forodesine
SA481235p_WT_6_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-6𝝰/Forodesine
SA481236WT_6_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-6𝝰/Forodesine
SA481237p_WT_7_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-7𝝰/Forodesine
SA481238WT_7_CytokinePlusForomouse primary astrocytes Wild-type TNF𝝰/IL-7𝝰/Forodesine
SA481239WT_1_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481240p_WT_6_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481241p_WT_5_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481242p_WT_4_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481243p_WT_3_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481244p_WT_2_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481245WT_2_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481246WT_6_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481247WT_5_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481248WT_4_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481249WT_3_Ctrlmouse primary astrocytes Wild-type Vehicle
SA481250p_WT_1_Ctrlmouse primary astrocytes Wild-type Vehicle
Showing results 1 to 80 of 80

Collection:

Collection ID:CO004308
Collection Summary:Primary astrocytes were isolated from the brains of wild-type or ENT2-/- mice at postnatal day 1 (P1). On DIV27, cells were stimulated with a cytokine cocktail, and metabolites were extracted on DIV28 using 0.6N perchloric acid (PCA).
Collection Protocol Filename:culture_and_collection_protocol.pdf
Sample Type:Astrocyte cells

Treatment:

Treatment ID:TR004324
Treatment Summary:Ent2+/+ and Ent2-/- primary astrocytes were treated on DIV27 with TNFα (30 ng/mL) and IL-1α (3 ng/mL) for 24 hours to induce activation. To inhibit PNP activity, Forodesine (2 μM) was applied simultaneously with cytokine treatment.
Treatment Protocol Filename:Treatment_protocol_24h.pdf

Sample Preparation:

Sampleprep ID:SP004321
Sampleprep Summary:Primary astrocytes were washed, digested, and subjected to metabolite extraction using 0.6 N perchloric acid (PCA), followed by neutralization with K₂CO₃. The clarified supernatants were collected and used for metabolomic profiling. The remaining protein pellets were processed separately and retained for concentration measurement.
Sampleprep Protocol Filename:PCA_extraction.pdf
Extract Storage:-80℃

Chromatography:

Chromatography ID:CH005250
Instrument Name:Agilent 1290 Infinity II UPLC system
Column Name:ACQUITY UPLC BEH Amide (100 x 2.1 mm, 1.7 μm)
Column Temperature:40℃
Flow Gradient:0–8 min, 90-50% B; 8–10 min, 50% B; 10–11 min, 50–90% B; 11–16 min, 90% B. (amide column)
Flow Rate:0.3 mL/min
Solvent A:100% Double-distilled Water; 15 mM ammonium acetate; 0.3% Ammonium hydroxide
Solvent B:90% Acetonitrile/10% Double-distilled Water; 15 mM ammonium acetate; 0.3% Ammonium hydroxide
Chromatography Type:HILIC

Analysis:

Analysis ID:AN006912
Analysis Type:MS
Analysis Protocol File:UPLC-MS-protocol.pdf
Chromatography ID:CH005250
Num Factors:29
Num Metabolites:31
Units:ion counts
  
Analysis ID:AN006913
Analysis Type:MS
Analysis Protocol File:UPLC-MS-protocol.pdf
Chromatography ID:CH005250
Num Factors:29
Num Metabolites:66
Units:ion counts
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