Summary of Study ST002740
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 PR001705. The data can be accessed directly via it's Project DOI: 10.21228/M89T4G 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 | ST002740 |
| Study Title | Non-targeted metabolomics screen comparing 13C2-acetate labeling of metabolites in CD8+ T cells and NK cells from mouse spleens. |
| Study Summary | Non-targeted metabolomics screen comparing 13C2-acetate labeling of metabolites in CD8+ T cells and NK cells from mouse spleens (wild type vs ACSS2 knockout C57Bl/6 mice). Metabolites were analyzed using a high-resolution, high-performance LC-MS analysis. |
| Institute | The Wistar Institute |
| Last Name | Schug |
| First Name | Zachary |
| Address | 3601 Spruce St, Philadelphia PA 19104 |
| zschug@wistar.org | |
| Phone | 215-898-3705 |
| Submit Date | 2023-06-20 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-07-11 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001705 |
| Project DOI: | doi: 10.21228/M89T4G |
| Project Title: | Acetate acts as a metabolic immunomodulator that potentiates anti-tumour immunity in breast cancer |
| Project Summary: | Acetate metabolism is an important metabolic pathway in many cancers and is controlled by acetyl-CoA synthetase 2 (ACSS2), an enzyme that catalyzes the conversion of acetate to acetyl-CoA. While the metabolic role of ACSS2 in cancer is well described, the consequences of blocking tumour acetate metabolism on the tumour microenvironment and anti-tumour immunity are unknown. We demonstrate that blocking ACSS2 switches cancer cells from acetate consumers to producers of acetate thereby freeing acetate for tumour-infiltrating lymphocytes to use as a fuel source. We show that acetate supplementation metabolically bolsters T-cell effector functions and proliferation. Targeting ACSS2 with CRISPR-Cas9 guides or a small molecule inhibitor promotes an anti-tumour immune response and enhances the efficacy of chemotherapy in preclinical breast cancer models. We propose a novel paradigm for targeting acetate metabolism in cancer in which inhibition of ACSS2 dually acts to impair tumour cell metabolism and potentiate anti-tumour immunity. |
| Institute: | The Wistar Institute |
| Last Name: | Schug |
| First Name: | Zachary |
| Address: | 3601 Spruce St, Philadelphia PA 19104 |
| Email: | zschug@wistar.org |
| Phone: | 215-898-3705 |
Subject:
| Subject ID: | SU002847 |
| 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 | Mouse model | Cell Type | [13C2-acetate] in mM | Time (hours) | ACSS2 inhibitor (Y/N) |
|---|---|---|---|---|---|---|
| SA288812 | Sample_KO1-100_3 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288813 | Sample_KO1-100_2 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288814 | Sample_KO1-100_1 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288815 | Sample_KO2-100_1 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288816 | Sample_KO2-100_2 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288817 | Sample_KO2-100_3 | Acss2-/- C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288818 | Sample_KO1-500_1 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288819 | Sample_KO1-500_2 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288820 | Sample_KO1-500_3 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288821 | Sample_KO2-500_3 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288822 | Sample_KO2-500_2 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288823 | Sample_KO2-500_1 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288824 | Sample_KO2-500_IN_3 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288825 | Sample_KO2-500_IN_2 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288826 | Sample_KO2-500_IN_1 | Acss2-/- C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288827 | Sample_1-100_1 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288828 | Sample_WT2-100_2 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288829 | Sample_1-100_3 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288830 | Sample_WT2-100_3 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288831 | Sample_1-100_2 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288832 | Sample_WT2-100_1 | WT C57Bl/6 | CD8+ T cells | 100 | 1 | N |
| SA288833 | Sample_2-100_3 | WT C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288834 | Sample_2-100_1 | WT C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288835 | Sample_2-100_2 | WT C57Bl/6 | CD8+ T cells | 100 | 2 | N |
| SA288836 | Sample_1-500_1 | WT C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288837 | Sample_1-500_3 | WT C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288838 | Sample_1-500_2 | WT C57Bl/6 | CD8+ T cells | 500 | 1 | N |
| SA288839 | Sample_WT2-500_3 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288840 | Sample_WT2-500_1 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288841 | Sample_2-500_2 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288842 | Sample_2-500_1 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288843 | Sample_WT2-500_2 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288844 | Sample_2-500_3 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | N |
| SA288845 | Sample_WT2-500-IN_3 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288846 | Sample_WT2-500-IN_2 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288847 | Sample_WT2-500-IN_1 | WT C57Bl/6 | CD8+ T cells | 500 | 2 | Y |
| SA288848 | Sample_N3-100_1 | WT C57Bl/6 | NK cells | 100 | 1 | N |
| SA288849 | Sample_N3-100_2 | WT C57Bl/6 | NK cells | 100 | 1 | N |
| SA288850 | Sample_N3-100_3 | WT C57Bl/6 | NK cells | 100 | 1 | N |
| SA288851 | Sample_N3-500_1 | WT C57Bl/6 | NK cells | 500 | 2 | N |
| SA288852 | Sample_N3-500_2 | WT C57Bl/6 | NK cells | 500 | 2 | N |
| SA288853 | Sample_N3-500_3 | WT C57Bl/6 | NK cells | 500 | 2 | N |
| Showing results 1 to 42 of 42 |
Collection:
| Collection ID: | CO002840 |
| Collection Summary: | CD8+ T cells and NK cells were purified by negative selection from freshly prepared mouse splenocytes (wild type vs ACSS2 knockout C57Bl/6 mice). |
| Sample Type: | Splenocytes |
Treatment:
| Treatment ID: | TR002856 |
| Treatment Summary: | Cells were incubated up to 3 hours in the in 100 or 500 µM 13C2-acetate and treated with vehicle (DMSO) or an ACSS2 inhibitor (VY-3-135). |
Sample Preparation:
| Sampleprep ID: | SP002853 |
| Sampleprep Summary: | Cells were pelleted and washed with ice-cold PBS before extraction of metabolites in 80% methanol. Samples were centrifuged at 4 °C two times (18,000 x g). Deproteinated supernatants were stored at -80 °C prior to analysis. |
Combined analysis:
| Analysis ID | AN004443 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish |
| Column | SeQuant ZIC-pHILIC (150 x 2.1mm, 5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH003338 |
| Chromatography Summary: | Hydrophilic interaction liquid chromatography (HILIC) was performed at 0.2 ml/min on a ZIC-pHILIC column (150 x 2.1 mm, 5 µM particle size, EMD Millipore) with a ZIC-pHILIC guard column (20 x 2.1 mm, EMD Millipore) at 30 °C. Solvent A was 20 mM ammonium carbonate, 0.1% ammonium hydroxide, pH 9.2, 5 µM medronic acid, and solvent B was acetonitrile. The gradient was 85% B for 2 min, 85% B to 20% B over 15 min, 20% B to 85% B over 0.1 min, and 85% B for 8.9 min. The autosampler was held at 4 °C. For each analysis, 5 µl of sample was injected. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm, 5um) |
| Column Temperature: | 30 |
| Flow Gradient: | 85% B for 2 min, 85% B to 20% B over 15 min, 20% B to 85% B over 0.1 min, and 85% B for 8.9 min |
| Flow Rate: | 0.2 ml/min |
| Solvent A: | 20 mM ammonium carbonate, 0.1% ammonium hydroxide, pH 9.2, 5 µM medronic acid |
| Solvent B: | acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004190 |
| Analysis ID: | AN004443 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The following parameters were used for the MS analysis: sheath gas flow rate, 30; auxiliary gas flow rate, 5; sweep gas flow rate, 0; auxiliary gas heater temperature, 200 °C; spray voltage, 3.6 kV for positive and negative modes; capillary temperature, 325 °C; and funnel RF level, 65. Samples were analyzed by full MS with polarity switching. Full MS scans were acquired at 60,000 resolution with 2 microscans and a scan range of 72-1080 m/z, automatic gain control (AGC) target of 5e6, and maximum injection time (IT) of 200 ms. |
| Ion Mode: | UNSPECIFIED |
