Summary of Study ST004027
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 PR002519. The data can be accessed directly via it's Project DOI: 10.21228/M81N95 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 | ST004027 |
| Study Title | Species-specific serine metabolism differentially controls natural killer cell functions |
| Study Summary | Primary PBMC-derived human NK cells and splenic mouse NK cells were isolated and either immediately extracted for metabolites or cultured for 3 days in IL-2/15 before metabolite extraction. Cells were rinsed with ice-cold 150 mM NH4AcO, pH 7.3, resuspended in 80% ice-cold MeOH extraction buffer, and centrifuged for 10 min at 17,000 g. Supernatants were collected and evaporated using a Nitrogen evaporator (Organomation). Evaporated extracts were stored at -80°C. Dried metabolites were re-suspended in 100 µL (1:1) dH20, ACN solution. 10% of these suspensions were injected per analysis. Samples were run on a Vanquish (Thermo Scientific) UHPLC system with mobile phase A (20 mM ammonium carbonate, pH 9.7) and mobile phase B (100% ACN) at a flow rate of 150 µL/min on a SeQuant ZIC-pHILIC Polymeric column (2.1 × 150 mm x 5 μm, EMD Millipore) at 35°C. Separation is achieved with a linear gradient from 20% A to 80% A in 20 min followed by a linear gradient from 80% A to 20% A from 20 min to 20.5 min. 20% A is then held from 20.5 min to 28 min. The UHPLC is coupled to a Q-Exactive (Thermo Scientific) mass analyzer running in polarity switching mode at 3.2 kV with an MS1 resolution of 70,000. |
| Institute | University of California, Los Angeles |
| Last Name | Li |
| First Name | Joey |
| Address | 615 Charles E Young Dr S, Los Angeles CA 90095 |
| joeyhli@g.ucla.edu | |
| Phone | 4086213407 |
| Submit Date | 2025-06-12 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-09-01 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002519 |
| Project DOI: | doi: 10.21228/M81N95 |
| Project Title: | Species-specific serine metabolism differentially controls natural killer cell functions |
| Project Summary: | Immune cells undergo rapid metabolic reprogramming to fuel effector responses. However, whether the metabolic pathways that supply these functions differ between human and mouse immune cells is poorly understood. Using a comparative metabolomics approach, here we show both conserved and species-distinct metabolite alterations in cytokine-activated primary human and mouse NK cells. Activated human NK cells fail to perform de novo serine synthesis, resulting in broadly impaired effector functions when serine starved ex vivo or during in vivo dietary serine restriction, limiting their antitumor function. In contrast, activated mouse NK cells perform de novo serine synthesis to fuel one-carbon metabolism and proliferation, resulting in increased metabolic flexibility during ex vivo and dietary serine restriction. While NK cells from both species require one-carbon metabolism to proliferate and produce interferon-gamma (IFN-γ), GCLC-dependent glutathione synthesis tunes cytotoxic versus inflammatory function in human NK cells. Thus, activated NK cell functions display species-specific requirements for serine metabolism, and environmental serine availability dictates activated human NK cell functions. |
| Institute: | University of California, Los Angeles |
| Department: | MIMG |
| Last Name: | Li |
| First Name: | Joey |
| Address: | 615 Charles E Young Dr S, Los Angeles CA 90095 |
| Email: | joeyhli@g.ucla.edu |
| Phone: | 4086213407 |
Subject:
| Subject ID: | SU004173 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens; Mus musculus |
| Taxonomy ID: | 9606 (Homo sapiens); 10090 (Mus musculus) |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens; Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Stimulation |
|---|---|---|---|
| SA464580 | Human_IL-215_1 | Human PBMC | 3 day IL-2/15 |
| SA464581 | Human_IL-215_2 | Human PBMC | 3 day IL-2/15 |
| SA464582 | Human_IL-215_3 | Human PBMC | 3 day IL-2/15 |
| SA464583 | Human_Naive_1 | Human PBMC | Naïve |
| SA464584 | Human_Naive_2 | Human PBMC | Naïve |
| SA464585 | Human_Naive_3 | Human PBMC | Naïve |
| SA464586 | Mouse_IL-215_1 | Mouse spleen | 3 day IL-2/15 |
| SA464587 | Mouse_IL-215_2 | Mouse spleen | 3 day IL-2/15 |
| SA464588 | Mouse_IL-215_3 | Mouse spleen | 3 day IL-2/15 |
| SA464589 | Mouse_Naive_1 | Mouse spleen | Naïve |
| SA464590 | Mouse_Naive_2 | Mouse spleen | Naïve |
| SA464591 | Mouse_Naive_3 | Mouse spleen | Naïve |
| Showing results 1 to 12 of 12 |
Collection:
| Collection ID: | CO004166 |
| Collection Summary: | Cells were rinsed with ice-cold 150 mM NH4AcO, pH 7.3, resuspended in 80% ice-cold MeOH extraction buffer, and centrifuged for 10 min at 17,000 g. Supernatants were collected and evaporated using a Nitrogen evaporator (Organomation). Evaporated extracts were stored at -80°C. |
| Sample Type: | Mononuclear cells; Spleen |
Treatment:
| Treatment ID: | TR004182 |
| Treatment Summary: | For cytokine stimulation, 2 x 104 mouse NK cells were cultured for 4 h at 37°C in CR-10 medium (RPMI 1640 + 10% FBS, 1% L-glutamine, 1% 200 mM sodium pyruvate, 1% MEM-NEAA, 1% penicillin-streptomycin, 0.5% sodium bicarbonate, and 0.01% 55 mM 2-mercaptoethanol), recombinant mouse IL-2 (100 IU/mL), and recombinant mouse IL-15 (50 ng/mL, PeproTech). Isolated human NK cells were cultured in 24-well G-Rex plates (Wilson Wolf) in NK MACS medium (Miltenyi Biotech) supplemented with recombinant human IL-2 (100 IU/mL, PeproTech) and recombinant human IL-15 (20 ng/mL, PeproTech). |
Sample Preparation:
| Sampleprep ID: | SP004179 |
| Sampleprep Summary: | Cells were rinsed with ice-cold 150 mM NH4AcO, pH 7.3, resuspended in 80% ice-cold MeOH extraction buffer, and centrifuged for 10 min at 17,000 g. Supernatants were collected and evaporated using a Nitrogen evaporator (Organomation). Evaporated extracts were stored at -80°C. Dried metabolites were re-suspended in 100 µL (1:1) dH20, ACN solution. |
Chromatography:
| Chromatography ID: | CH005056 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | SeQuant ZIC- pHILIC (150 x 2.1 mm, 5 µm) |
| Column Temperature: | 35°C |
| Flow Gradient: | 0-20 min: 20%-80%; 20-20.5 min: 80%-20%; 20.5-28 min: 20% |
| Flow Rate: | 150 µL/min |
| Solvent A: | 100% Water; 20 mM Ammonium carbonate, pH 9.7 |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN006653 |
| Analysis Type: | MS |
| Chromatography ID: | CH005056 |
| Num Factors: | 4 |
| Num Metabolites: | 27 |
| Units: | normalized intensity |
| Analysis ID: | AN006654 |
| Analysis Type: | MS |
| Chromatography ID: | CH005056 |
| Num Factors: | 4 |
| Num Metabolites: | 55 |
| Units: | normalized intensity |
