Summary of Study ST003571
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 | ST003571 |
| Study Title | Hepatocytes transform fructose into lipids that can metabolized by cancer cells |
| Study Summary | Fructose is known to be metabolized by the liver to produce lipids that can circulate in the systemic circulation. Metabolism of fructose by CaSki cancer cells, on the other hand, is minimal. Therefore, we wished to understand if fructose metabolism in hepatocytes resulted in the release of lipids that CaSki cells could then uptake and metabolize to support their growth. Therefore, we formed fresh media which contained fructose as the only sugar source in the media. We cultured primary hepatocytes from C57BL/6 mice in this fresh media to form hepatocyte-conditioned media. We then transferred this hepatocyte-conditioned media to CaSki cells to form CaSki-conditioned media. LC-MS analysis on reverse-phase chromatography of fresh media, hepatocyte-conditioned media, and CaSki-conditioned media revealed that several lysophosphatidylcholine species are released from hepatocytes and 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 | 2024-11-10 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-11-27 |
| 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: | SU003700 |
| 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 |
|---|---|---|---|
| SA389609 | Caski CM 2 | culture media | CaSki cells |
| SA389610 | Caski CM 1 | culture media | CaSki cells |
| SA389611 | Caski CM 3 | culture media | CaSki cells |
| SA389612 | Caski CM 4 | culture media | CaSki cells |
| SA389613 | hepatocyte CM 1 | culture media | mouse primary hepatocytes |
| SA389614 | hepatocyte CM 2 | culture media | mouse primary hepatocytes |
| SA389615 | hepatocyte CM 3 | culture media | mouse primary hepatocytes |
| SA389616 | hepatocyte CM 4 | culture media | mouse primary hepatocytes |
| SA389617 | hepatocyte CM 5 | culture media | mouse primary hepatocytes |
| SA389618 | hepatocyte CM 6 | culture media | mouse primary hepatocytes |
| SA389619 | Fresh media 1 | culture media | no cells |
| SA389620 | Fresh media 2 | culture media | no cells |
| SA389621 | Fresh media 3 | culture media | no cells |
| Showing results 1 to 13 of 13 |
Collection:
| Collection ID: | CO003693 |
| Collection Summary: | 10 mM fructose was dissolved in glucose-free DMEM and 10% dialyzed FBS with 1% penicillin/streptomycin. This media with no cells composed the fresh media. 50 uL of this media was then collected and later extracted for LC-MS analysis, composing the fresh media condition. The media was then used to culture hepatocytes for 24 hours, yielding hepatocyte conditioned media (CM). 50 uL of this media was collected and later extracted for LC-MS analysis, composing the hepatocyte conditioned media collection. This hepatocyte conditioned media was then transferred to CaSki cells for 24 hours, yielding Caski conditioned media (CM). 50 uL of this media was collected and later extracted for LC-MS analysis, composing the CaSki conditioned media. |
| Sample Type: | Culture Media |
Treatment:
| Treatment ID: | TR003709 |
| Treatment Summary: | 10 mM fructose was dissolved in glucose-free DMEM and 10% dialyzed FBS with 1% penicillin/streptomycin. This media with no cells composed the fresh media. The media was then used to culture hepatocytes for 24 hours, yielding hepatocyte conditioned media (CM). This hepatocyte conditioned media was then transferred to CaSki cells for 24 hours, yielding Caski conditioned media (CM). |
Sample Preparation:
| Sampleprep ID: | SP003707 |
| Sampleprep Summary: | For extractions of cell-culture media, 50 µL of serum was added to a Captiva EMR 96 well plate (Agilent, Santa Clara, CA). Acetonitrile:methanol (1:1, 200 µL) with labeled internal standards was added to the plate and incubated for 1 minute on a plate shaker and at 4 °C for 10 minutes. Methaol:acetonitrile:water (2:2:1, 150 µL) was added to the plate and eluted by using a positive pressure manifold into a collection plate. The Captiva EMR 96-well plate was washed one additional time with methanol:acetonitrile:water (2:2:1) and eluted for polar metabolites. For nonpolar metabolites, a new collection plate was used, and eluted with 1:1 methanol:methyl tert-butyl ether (MTBE). Eluted nonpolar metabolites were dried under a stream of N2 by a Biotage N2-dryer. Nonpolar metabolites were reconstituted in 1:1 isopropanol:methanol. A pooled-reference sample was prepared by mixing aliquots of all samples prior to extraction. |
Combined analysis:
| Analysis ID | AN005866 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 Infinity II |
| Column | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6545 QTOF |
| Ion Mode | POSITIVE |
| Units | Peak area |
Chromatography:
| Chromatography ID: | CH004455 |
| Chromatography Summary: | Samples were analyzed by using a HSS T3 column (Acquity; 150 × 2.1 mm, 1.8 μm) interfaced with an Agilent 6545 QTOF. The LC system used was an Agilent 1290 Infinity II. Mobile-phase solvents had the following composition: A = 60% acetonitrile, 40% water, 0.1% formic acid, 10 mM ammonium formate 2.5 μM medronic acid and B = 90% 2-propanol, 10% acetonitrile, 0.1% formic acid, 10 mM ammonium formate. The following linear gradient was used: 0-2 min, 30% B; 17 min, 75% B; 20 min, 85% B; 23-26 min, 100% B; 26 min, 30% B. Injection volumes were 4 μL. The column compartment was maintained at 60 °C |
| Instrument Name: | Agilent 1290 Infinity II |
| Column Name: | Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) |
| Column Temperature: | 60 |
| Flow Gradient: | 0-2 min, 30% B; 17 min, 75% B; 20 min, 85% B; 23-26 min, 100% B; 26 min, 30% B |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 60% acetonitrile/40% water; 0.1% formic acid; 10 mM ammonium formate; 2.5 μM medronic acid |
| Solvent B: | 90% 2-propanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005586 |
| Analysis ID: | AN005866 |
| Instrument Name: | Agilent 6545 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | LC/MS of nonpolar metabolites was performed by using a HSS T3 column (Acquity; 150 × 2.1 mm, 1.8 μm) interfaced with an Agilent 6545 Q-TOF. The mass range was 120-1200 m/z. Instrument parameters were as follows: gas, 250°C at 11 L/min; nebulizer pressure, 35 psi; sheath gas temperature, 300°C; sheath gas flow 12 L/min; VCap 3000 V; nozzle voltage 500 V; Fragmentor 160 V; Skimmer 65 V. The instrument was operated in positive ionization mode for all samples analyzed. |
| Ion Mode: | POSITIVE |
