Summary of Study ST003572

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.

Study ID	ST003572
Study Title	Dietary sugar supplementation in mice enhances the availability of many circulating lipids including lysophosphatidylcholines
Study Summary	Sugar consumption is known to enhance de novo lipogenesis, but it is still not comprehensively known which lipid species are altered secondary to sugar consumption. To understand the effects of sugar consumption on circulating lipids, we supplemented C57BL/6 mice on a modified chow diet that contained no free sugars with either normal water, 10% glucose water, 10% fructose water, or 20% HFCS water. A comparative analysis of the serum between the four groups revealed 128 lipids that had statistically significant changes among all four conditions. We observed that LPCs were a particularly altered lipid class among the lipids we detected. Notably, 16:1 and 18:1 LPCs were increased severalfold in either the 10% glucose, 10% fructose, or 20% HFCS conditions relative to the normal water condition.
Institute	Washington University in St. Louis
Department	Chemistry
Laboratory	Gary Patti
Last Name	Fowle-Grider
First Name	Ronald
Address	5630 Pershing Ave
Email	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

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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:	SU003701
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Species Group:	Mammals

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id	local_sample_id	Sample source	Mouse dietary treatment
SA389622	fructose serum 1	Mouse serum	fructose water
SA389623	fructose serum 5	Mouse serum	fructose water
SA389624	fructose serum 4	Mouse serum	fructose water
SA389625	fructose serum 2	Mouse serum	fructose water
SA389626	fructose serum 3	Mouse serum	fructose water
SA389627	Glucose serum 3	Mouse serum	glucose water
SA389628	Glucose serum 4	Mouse serum	glucose water
SA389629	Glucose serum 2	Mouse serum	glucose water
SA389630	Glucose serum 1	Mouse serum	glucose water
SA389631	HFCS serum 1	Mouse serum	high fructose corn syrup (HFCS) water
SA389632	HFCS serum 2	Mouse serum	high fructose corn syrup (HFCS) water
SA389633	HFCS serum 3	Mouse serum	high fructose corn syrup (HFCS) water
SA389634	HFCS serum 4	Mouse serum	high fructose corn syrup (HFCS) water
SA389635	HFCS serum 5	Mouse serum	high fructose corn syrup (HFCS) water
SA389636	HFCS serum 6	Mouse serum	high fructose corn syrup (HFCS) water
SA389637	Control serum 2	Mouse serum	normal water
SA389638	Control serum 4	Mouse serum	normal water
SA389639	Control serum 3	Mouse serum	normal water
SA389640	Control serum 1	Mouse serum	normal water

Showing results 1 to 19 of 19

Showing results 1 to 19 of 19

Collection:

Collection ID:	CO003694
Collection Summary:	Whole blood was obtained from mice in all conditions at the endpoint of the experiment via cardiac puncture and mice were sacrificed. Collected whole-blood was placed on ice without anticoagulant for 20 minutes. The samples were subsequently centrifuged at 6000 rpm for 10 minutes. Serum was then collected and stored at -80 ºC until extraction for LC/MS analysis.
Sample Type:	Blood (serum)

Treatment:

Treatment ID:	TR003710
Treatment Summary:	We used LC/MS to profile lipid metabolites from the serum of mice on high-sugar diets. For a control, mice were given regular facility water and fed a diet resembling normal chow but without any fructose or free sugars. Animals on high-sugar diets were fed the same chow but supplemented with 10% glucose water, 10% fructose water, or 20% HFCS water. All mice were on the indicated dietary regimes for 6 weeks, after which mice were sacrificed and serum was obtained for LC-MS analysis. The control group on normal water contain n=4 mice; the 10% glucose water group had n=4 mice; the 10% fructose group had n=5 mice; the HFCS group had n=6 mice. Each LC-MS sample analyzed represents the serum from just one mouse of the indicated treatment group.

Treatment ID:

TR003710

Treatment Summary:

We used LC/MS to profile lipid metabolites from the serum of mice on high-sugar diets. For a control, mice were given regular facility water and fed a diet resembling normal chow but without any fructose or free sugars. Animals on high-sugar diets were fed the same chow but supplemented with 10% glucose water, 10% fructose water, or 20% HFCS water. All mice were on the indicated dietary regimes for 6 weeks, after which mice were sacrificed and serum was obtained for LC-MS analysis. The control group on normal water contain n=4 mice; the 10% glucose water group had n=4 mice; the 10% fructose group had n=5 mice; the HFCS group had n=6 mice. Each LC-MS sample analyzed represents the serum from just one mouse of the indicated treatment group.

Sample Preparation:

Sampleprep ID:	SP003708
Sampleprep Summary:	For extractions of mouse serum, 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.

Sampleprep ID:

SP003708

Sampleprep Summary:

For extractions of mouse serum, 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	AN005867
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:	CH004456
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:	MS005587
Analysis ID:	AN005867
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