Summary of Study ST002935
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 PR001826. The data can be accessed directly via it's Project DOI: 10.21228/M8PH9P 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 | ST002935 |
| Study Title | Title: Myeloid cell-derived creatine in the hypoxic niche promotes glioblastoma growth |
| Study Type | PBMC vs Tumor CD163+ |
| Study Summary | Glioblastoma (GBM) is a malignancy dominated by the infiltration of tumor-associated myeloid cells (TAMCs). Examination of TAMC metabolic phenotypes in mouse models and human GBM patients identified the de-novo creatine metabolic pathway as a hallmark of TAMCs. Multi-omics analyses revealed that TAMCs surround the hypoxic peri-necrotic regions of GBM and express the creatine metabolic enzyme glycine amidinotransferase (GATM). Conversely, GBM cells located within these same regions are uniquely specific in expressing the creatine transporter (SLC6A8). Therefore, we hypothesized that TAMCs provide creatine to tumors, promoting GBM progression. Isotopic tracing demonstrated that TAMC-secreted creatine can be taken up by tumor cells. Creatine supplementation protected tumors from hypoxia-induced stress which was abrogated with genetic ablation or pharmacologic inhibition of SLC6A8. Lastly, inhibition of creatine transport using the clinically relevant compound, RGX-202-01, blunted tumor growth, and enhanced radiation therapy in-vivo. This work highlights that myeloid-to-tumor transfer of creatine promotes tumor growth in the hypoxic niche. |
| Institute | Northwestern University, Feinberg School of Medicine |
| Department | Neurological Surgery |
| Laboratory | Jason Miska |
| Last Name | Miska |
| First Name | Jason |
| Address | 676 N St. Clair |
| jason.miska@northwestern.edu | |
| Phone | 8478678201 |
| Submit Date | 2023-10-16 |
| Num Groups | 2 |
| Total Subjects | 5 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-11-03 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001826 |
| Project DOI: | doi: 10.21228/M8PH9P |
| Project Title: | Myeloid cell-derived creatine in the hypoxic niche promotes glioblastoma growth |
| Project Type: | LC-MS/MS |
| Project Summary: | Glioblastoma (GBM) is a malignancy dominated by the infiltration of tumor-associated myeloid cells (TAMCs). Examination of TAMC metabolic phenotypes in mouse models and human GBM patients identified the de-novo creatine metabolic pathway as a hallmark of TAMCs. Multi-omics analyses revealed that TAMCs surround the hypoxic peri-necrotic regions of GBM and express the creatine metabolic enzyme glycine amidinotransferase (GATM). Conversely, GBM cells located within these same regions are uniquely specific in expressing the creatine transporter (SLC6A8). Therefore, we hypothesized that TAMCs provide creatine to tumors, promoting GBM progression. Isotopic tracing demonstrated that TAMC-secreted creatine can be taken up by tumor cells. Creatine supplementation protected tumors from hypoxia-induced stress which was abrogated with genetic ablation or pharmacologic inhibition of SLC6A8. Lastly, inhibition of creatine transport using the clinically relevant compound, RGX-202-01, blunted tumor growth, and enhanced radiation therapy in-vivo. This work highlights that myeloid-to-tumor transfer of creatine promotes tumor growth in the hypoxic niche. |
| Institute: | Northwestern University, Feinberg School of Medicine |
| Department: | Neurological Surgery |
| Laboratory: | Jason Miska |
| Last Name: | Miska |
| First Name: | Jason |
| Address: | 676 N St. Clair |
| Email: | jason.miska@northwestern.edu |
| Phone: | 8478678201 |
Subject:
| Subject ID: | SU003048 |
| 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 |
|---|---|---|
| SA318402 | Les-Jas-20190812-13 | PBMC |
| SA318403 | Les-Jas-20200214-04 | PBMC |
| SA318404 | Les-Jas-20190812-14 | PBMC |
| SA318405 | Les-Jas-20190812-15 | PBMC |
| SA318406 | Les-Jas-20200214-03 | PBMC |
| SA318407 | Les-Jas-20200214-02 | Tumor |
| SA318408 | Les-Jas-20190812-18 | Tumor |
| SA318409 | Les-Jas-20190812-16 | Tumor |
| SA318410 | Les-Jas-20190812-17 | Tumor |
| SA318411 | Les-Jas-20200214-01 | Tumor |
| Showing results 1 to 10 of 10 |
Collection:
| Collection ID: | CO003041 |
| Collection Summary: | Magnetic bead isolation of cells To isolate specific populations of cells, single-cell suspensions as isolated above are preblocked with anti-CD16/32 for 15 min at 4°C. We then used the biotinylated anti-CD163 (all from Thermo Fisher Scientific) to label murine myeloid cells. Next, the cells were washed and then incubated with anti-biotin magnetic beads (Miltenyi Biotec) before performing manual positive selection using MS columns (Miltenyi Biotec). Purified cells were analyzed for all downstream metabolic analyses. |
| Sample Type: | Brain |
Treatment:
| Treatment ID: | TR003057 |
| Treatment Summary: | Fresh cells isolated from PBMC or TIL of GBM patients |
Sample Preparation:
| Sampleprep ID: | SP003054 |
| Sampleprep Summary: | Isolated TAMC samples were dried using a SpeedVac. Acetonitrile (50%) was added to the tube for reconstitution following overtaxing for 30 s. Sample solution was then centrifuged for 15 min at 20,000g and 4°C. Supernatant was collected for LC-MS analysis. |
Combined analysis:
| Analysis ID | AN004814 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Water's Xbridge amide (100 x 3mm, 3.5 um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Peak Area |
Chromatography:
| Chromatography ID: | CH003638 |
| Chromatography Summary: | Samples were analyzed by high-performance LC (HPLC) and high-resolution MS and MS/MS (HPLC-MS/MS). The system consists of Thermo Q Exactive with an electrospray source and an UltiMate3000 (Thermo Fisher Scientific) series HPLC consisting of a binary pump, degasser, and autosampler outfitted with an XBridge Amide column (Waters; dimensions of 4.6 mm by 100 mm and a 3.5-μm particle size). The mobile phase A contained 95% water/5% acetonitrile (v/v), 20 mM ammonium hydroxide, and 20 mM ammonium acetate (pH 9.0); phase B was 100% acetonitrile. The gradient was performed as follows: 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A with a flow rate of 400 µl/min. |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Water's Xbridge amide (100 x 3mm, 3.5 um) |
| Column Temperature: | - |
| Flow Gradient: | 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A |
| Flow Rate: | 400 μl/min |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium hydroxide; 20 mM ammonium acetate (pH 9.0) |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004560 |
| Analysis ID: | AN004814 |
| Instrument Name: | Thermo Q Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The capillary of the electrospray ionization source was set to 275°C, with sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units, and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, a mass/charge ratio (m/z) scan range from 70 to 850 was chosen and MS1 data were collected at a resolution of 70,000. The automatic gain control target was set at 1 × 106, and the maximum injection time was 200 ms. The top five precursor ions were subsequently fragmented, in a data-dependent manner, using the higher-energy collisional dissociation cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500. |
| Ion Mode: | UNSPECIFIED |
