Summary of Study ST003570
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 PR002203. The data can be accessed directly via it's Project DOI: 10.21228/M8VN7V 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 ID | ST003570 |
| Study Title | Hepatocytes transform fructose into polar metabolites that are selectively utilized by cancer cells |
| Study Summary | The goal of this project was to determine if hepatocytes can metabolically transform fructose into polar metabolites that CaSki cancer cells can uptake and metabolize. The CaSki cells do not metabolize fructose directly to much extent, but the liver robustly metabolizes fructose. Therefore, we cultured primary hepatocytes from C57BL/6 mice in [U-13C] fructose as the only sugar source for 24 hours to form hepatocyte-conditioned media. We then transferred this media to CaSki cells for 24 hours to form CaSki-conditioned media. LC-MS profiling of polar 13C-labeled metabolites of hepatocyte-conditioned media and CaSki-conditioned media revealed that 13C-labeled glucose and 13C-labeled aspartate are released from hepatocytes and are up taken by CaSki cells. |
| Institute | Washington University in St. Louis |
| Department | Chemistry |
| Laboratory | Gary Patti |
| Last Name | Fowle-Grider |
| First Name | Ronald |
| Address | 5630 Pershing Ave |
| rjfowle@wustl.edu | |
| Phone | 3092657545 |
| Submit Date | 2024-10-31 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-03-17 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002203 |
| Project DOI: | doi: 10.21228/M8VN7V |
| Project Title: | Dietary fructose enhances tumor growth indirectly via interorgan lipid transfer |
| Project Summary: | Fructose consumption has increased considerably over the past five decades, largely due to the widespread use of high-fructose corn syrup (HFCS) as a sweetener. It has been proposed that fructose promotes the growth of some tumors by serving as a direct fuel. Here, we show that fructose supplementation enhances tumor growth in animal models of melanoma, breast cancer, and cervical cancer without causing weight gain or insulin resistance. Interestingly, the cancer cells themselves were unable to use fructose readily as a nutrient because they did not express ketohexokinase-C (KHK-C). Primary hepatocytes did express KHK-C, resulting in fructolysis and the excretion of a variety of lipid species, including lysophosphatidylcholines (LPCs). In co-culture experiments, hepatocyte-derived LPCs were consumed by cancer cells and used to generate phosphatidylcholines (PCs), the major phospholipid of cell membranes. In vivo, HFCS supplementation increased several LPC species by >7-fold in serum. Administration of LPCs to mice was sufficient to increase tumor growth. Pharmacological inhibition of ketohexokinase had no direct effect on cancer cells, but it decreased circulating LPC levels and prevented fructose-mediated tumor growth in vivo. These findings reveal that fructose supplementation increases circulating nutrients such as LPCs, which can enhance tumor growth through a cell non-autonomous mechanism. |
| Institute: | Washington University in St. Louis |
| Department: | Chemistry |
| Laboratory: | Gary Patti |
| Last Name: | Fowle-Grider |
| First Name: | Ronald |
| Address: | 6101 Washington Blvd Unit 202, SAINT LOUIS, MO, 63112, USA |
| Email: | rjfowle@wustl.edu |
| Phone: | 309-265-7545 |
Subject:
| Subject ID: | SU003699 |
| Subject Type: | Cultured cells |
| Subject Species: | Human (Homo sapiens), Mouse (Mus musculus) |
| Species Group: | Mammals |
Factors:
Subject type: Cultured cells; Subject species: Human (Homo sapiens), Mouse (Mus musculus) (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Conditioned media cell type |
|---|---|---|---|
| SA389603 | CaSki CM C13 Fructose 1 | culture media | CaSki cells |
| SA389604 | CaSki CM C13 Fructose 2 | culture media | CaSki cells |
| SA389605 | CaSki CM C13 Fructose 3 | culture media | CaSki cells |
| SA389606 | Hepatocyte CM C13 Fructose 1 | culture media | Hepatocytes |
| SA389607 | Hepatocyte CM C13 Fructose 2 | culture media | Hepatocytes |
| SA389608 | Hepatocyte CM C13 Fructose 3 | culture media | Hepatocytes |
| Showing results 1 to 6 of 6 |
Collection:
| Collection ID: | CO003692 |
| Collection Summary: | 10 mM [U-13C] fructose was dissolved in glucose-free DMEM and 10% dialyzed FBS with 1% penicillin/streptomycin. The medium was used to culture hepatocytes for 24 hours, yielding hepatocyte-conditioned medium (CM). 50 uL of this hepatocyte-conditioned medium was then collected and placed in the -80 degree celsius freezer for subsequent extraction for LC-MS analysis. This hepatocyte-conditioned medium was then transferred to CaSki cells for 24 hours, yielding Caski-conditioned medium (CM).50 uL of this CaSki-conditioned medium was then collected and placed in the -80 degree celsius freezer for subsequent extraction for LC-MS analysis. |
| Sample Type: | Culture Media |
Treatment:
| Treatment ID: | TR003708 |
| Treatment Summary: | 10 mM [U-13C] fructose was dissolved in glucose-free DMEM and 10% dialyzed FBS with 1% penicillin/streptomycin. The medium was used to culture hepatocytes for 24 hours, yielding hepatocyte-conditioned medium (CM). This hepatocyte-conditioned medium was then transferred to CaSki cells for 24 hours, yielding Caski-conditioned media (CM). |
Sample Preparation:
| Sampleprep ID: | SP003706 |
| Sampleprep Summary: | 90 µL of 2:2:1 methanol:acetonitrile:water (M:A:W) was added to 10 µL of cell culture media sample. The samples were vortexed and placed at -20 ºC for an hour. The samples then were centrifuged at 4 ºC for 10 minutes. The supernatant was removed and analyzed by LC/MS. |
Combined analysis:
| Analysis ID | AN005865 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Agilent 1290 Infinity II |
| Column | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6540 QTOF |
| Ion Mode | NEGATIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH004454 |
| Chromatography Summary: | LC/MS of polar metabolites was performed by using a SeQuant ZIC-pHILIC column (EMD Millipore) interfaced with an Agilent 6540 Q-TOF. A 150 × 2.1 mm, 5 μm SeQuant ZIC-pHILIC column was used with an Agilent 1290 Infinity II LC system, applying methods established previously 65. Mobile-phase solvents had the following composition: A = 20 mM ammonium acetate in water:acetonitrile (95:5) and B = 100% acetonitrile. The following linear gradient was used: 0−0.5 min, 90% B; 0.5−30 min, 90−30% B; 30−31 min, 30% B. Injection volumes were 2 μL for all experiments. The column compartment was maintained at 45 °C. |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0−0.5 min, 90% B; 0.5−30 min, 90−30% B linear gradient; 30−31 min, 30% B |
| Flow Rate: | 0.2 mL/min |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005585 |
| Analysis ID: | AN005865 |
| Instrument Name: | Agilent 6540 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | LC/MS of polar metabolites was performed by using a SeQuant ZIC-pHILIC column (EMD Millipore) interfaced with an Agilent 6540 Q-TOF. The mass range was set to 50 to 1500 m/z. Instrument parameters were as follows: gas, 200 °C at 4 L/min; nebulizer, 44 psi at 2000 V; sheath gas, 300 °C at 12 L/min, capillary, 3000 V; fragmentor, 100 V; skimmer, 65 V; and scan rate, 3 scans/second. The instrument was operated in negative ionization mode for all samples analyzed. Isotopologues and 13C enrichment in metabolites of interest were generated from analysis of the raw data |
| Ion Mode: | NEGATIVE |
