Summary of Study ST002161

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 PR001374. The data can be accessed directly via it's Project DOI: 10.21228/M84127 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	ST002161
Study Title	Glycerate Production from Intestinal Fructose Metabolism Elevated by Dietary Fat Induces Glucose Intolerance Through β-cell Damage
Study Summary	Dietary fructose, especially in the context of a high-fat western diet, has been linked to type 2 diabetes. Although the effect of fructose on liver metabolism has been extensively studied, a significant portion of the fructose is first metabolized in the small intestine. Here we report that dietary fat enhances intestinal fructose metabolism, which releases glycerate into the blood. High systemic glycerate levels reduce pancreatic islet sizes and β-cell content, thus inducing glucose intolerance. Our findings provide an additional link between dietary fructose and diabetes that is modulated by dietary fat.
Institute	Duke University
Last Name	Wong
First Name	Chi Wut
Address	CEIMAS, 101 Science Dr. Room 2141, Durham, NC, 27709, USA
Email	chiwut.wong@duke.edu
Phone	9495290320
Submit Date	2022-05-11
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2022-07-06
Release Version	1

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Project:

Project ID:	PR001374
Project DOI:	doi: 10.21228/M84127
Project Title:	Glycerate Production from Intestinal Fructose Metabolism Elevated by Dietary Fat Induces Glucose Intolerance Through β-cell Damage
Project Summary:	Dietary fructose, especially in the context of a high-fat western diet, has been linked to type 2 diabetes. Although the effect of fructose on liver metabolism has been extensively studied, a significant portion of the fructose is first metabolized in the small intestine. Here we report that dietary fat enhances intestinal fructose metabolism, which releases glycerate into the blood. High systemic glycerate levels reduce pancreatic islet sizes and β-cell content, thus inducing glucose intolerance. Our findings provide an additional link between dietary fructose and diabetes that is modulated by dietary fat.
Institute:	Duke University
Last Name:	Wong
First Name:	Chi Wut
Address:	CEIMAS, 101 Science Dr. Room 2141, Durham, NC, 27709, USA
Email:	chiwut.wong@duke.edu
Phone:	9495290320

Subject:

Subject ID:	SU002247
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090

Factors:

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

mb_sample_id	local_sample_id	Genotype	Diet	Diet Duration	Treatment	Tissue	Collection Time (minute)	Drinking
SA207163	Blank_batch4	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207164	Blank_batch3	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207165	Blank_batch11	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207166	Blank_batch5	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207167	Blank_batch2	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207168	Blank_batch6	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207169	Blank_batch9	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207170	Blank_batch8	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207171	Blank_batch7	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207172	Blank_batch10	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207173	Blank_batch1	Blank	Blank	Blank	Blank	Blank	Blank	Blank
SA207177	KHK_Gly_5_010	KHK-c-KO	CD	3 days	Control	Systemic Serum	10	Water
SA207178	KHK_Gly_4_010	KHK-c-KO	CD	3 days	Control	Systemic Serum	10	Water
SA207179	KHK_Gly_1_010	KHK-c-KO	CD	3 days	Control	Systemic Serum	10	Water
SA207180	KHK_Gly_5_120	KHK-c-KO	CD	3 days	Control	Systemic Serum	120	Water
SA207181	KHK_Gly_4_120	KHK-c-KO	CD	3 days	Control	Systemic Serum	120	Water
SA207182	KHK_Gly_1_120	KHK-c-KO	CD	3 days	Control	Systemic Serum	120	Water
SA207183	KHK_Gly_5_030	KHK-c-KO	CD	3 days	Control	Systemic Serum	30	Water
SA207184	KHK_Gly_1_030	KHK-c-KO	CD	3 days	Control	Systemic Serum	30	Water
SA207185	KHK_Gly_4_030	KHK-c-KO	CD	3 days	Control	Systemic Serum	30	Water
SA207186	KHK_Gly_5_060	KHK-c-KO	CD	3 days	Control	Systemic Serum	60	Water
SA207187	KHK_Gly_1_060	KHK-c-KO	CD	3 days	Control	Systemic Serum	60	Water
SA207188	KHK_Gly_4_060	KHK-c-KO	CD	3 days	Control	Systemic Serum	60	Water
SA207174	KHK_Gly_4_000	KHK-c-KO	CD	3 days	Control	Systemic Serum	-	Water
SA207175	KHK_Gly_1_000	KHK-c-KO	CD	3 days	Control	Systemic Serum	-	Water
SA207176	KHK_Gly_5_000	KHK-c-KO	CD	3 days	Control	Systemic Serum	-	Water
SA207189	Unlabeled_LF-HV_6348	Wild-type	CD	12 weeks	Fructose injection	Systemic Serum	1440	Water
SA207190	Unlabeled_LF-HV_13628	Wild-type	CD	12 weeks	Fructose injection	Systemic Serum	1440	Water
SA207191	Unlabeled_LF-HV_13626	Wild-type	CD	12 weeks	Fructose injection	Systemic Serum	1440	Water
SA207192	Unlabeled_LF-HV_13631	Wild-type	CD	12 weeks	Fructose injection	Systemic Serum	1440	Water
SA207193	Unlabeled_LF-HV_13627	Wild-type	CD	12 weeks	Fructose injection	Systemic Serum	1440	Water
SA207194	Unlabeled_LH-HV_10632	Wild-type	CD	12 weeks	Glycerate injection	Systemic Serum	1440	Water
SA207195	Unlabeled_LH-HV_916	Wild-type	CD	12 weeks	Glycerate injection	Systemic Serum	1440	Water
SA207196	Unlabeled_LH-HV_10684	Wild-type	CD	12 weeks	Glycerate injection	Systemic Serum	1440	Water
SA207197	Unlabeled_LH-HV_914	Wild-type	CD	12 weeks	Glycerate injection	Systemic Serum	1440	Water
SA207198	Unlabeled_LH-HV_917	Wild-type	CD	12 weeks	Glycerate injection	Systemic Serum	1440	Water
SA207199	Unlabeled_LC-HV_13646	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207200	Unlabeled_LC-HV_13683	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207201	Unlabeled_LC-HV_13637	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207202	Unlabeled_LC-HV_13635	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207203	Unlabeled_LC-HV_13642	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207204	Unlabeled_LC-HV_13634	Wild-type	CD	12 weeks	Saline injection	Systemic Serum	1440	Water
SA207205	Jej-13172	Wild-type	CD	3 days	Control	Jejunum	10	Water
SA207206	Intestine_01	Wild-type	CD	3 days	Control	Jejunum	10	Water
SA207207	Jej-13170	Wild-type	CD	3 days	Control	Jejunum	10	Water
SA207208	Jej-13171	Wild-type	CD	3 days	Control	Jejunum	10	Water
SA207213	L12130_45_02	Wild-type	CD	3 days	Control	Pancreas	45	Water
SA207214	L12126_45_02	Wild-type	CD	3 days	Control	Pancreas	45	Water
SA207215	L12129_45_02	Wild-type	CD	3 days	Control	Pancreas	45	Water
SA207216	L12128_45_02	Wild-type	CD	3 days	Control	Pancreas	45	Water
SA207217	L12140_60_03	Wild-type	CD	3 days	Control	Pancreas	60	Water
SA207218	L12137_60_03	Wild-type	CD	3 days	Control	Pancreas	60	Water
SA207219	L12139_60_03	Wild-type	CD	3 days	Control	Pancreas	60	Water
SA207220	L12136_60_03	Wild-type	CD	3 days	Control	Pancreas	60	Water
SA207209	L13157_0_01	Wild-type	CD	3 days	Control	Pancreas	-	Water
SA207210	L13160_0_01	Wild-type	CD	3 days	Control	Pancreas	-	Water
SA207211	L13159_0_01	Wild-type	CD	3 days	Control	Pancreas	-	Water
SA207212	L13156_0_01	Wild-type	CD	3 days	Control	Pancreas	-	Water
SA207224	13C-Fruc_LFD-PV_13171	Wild-type	CD	3 days	Control	Portal Vein Serum	10	Water
SA207225	13C-Fruc_LFD-PV_13170	Wild-type	CD	3 days	Control	Portal Vein Serum	10	Water
SA207226	13C-Fruc_LFD-PV_13172	Wild-type	CD	3 days	Control	Portal Vein Serum	10	Water
SA207227	13C-Fruc_LFD-PV_13168	Wild-type	CD	3 days	Control	Portal Vein Serum	10	Water
SA207228	13C-Fruc_LFD-PV_13169	Wild-type	CD	3 days	Control	Portal Vein Serum	10	Water
SA207229	CON-PV_120_12156	Wild-type	CD	3 days	Control	Portal Vein Serum	120	Water
SA207230	CON-PV_120_12158	Wild-type	CD	3 days	Control	Portal Vein Serum	120	Water
SA207231	CON-PV_120_12157	Wild-type	CD	3 days	Control	Portal Vein Serum	120	Water
SA207232	CON-PV_15_12107	Wild-type	CD	3 days	Control	Portal Vein Serum	15	Water
SA207233	CON-PV_15_12106	Wild-type	CD	3 days	Control	Portal Vein Serum	15	Water
SA207234	CON-PV_15_12109	Wild-type	CD	3 days	Control	Portal Vein Serum	15	Water
SA207235	CON-PV_15_12110	Wild-type	CD	3 days	Control	Portal Vein Serum	15	Water
SA207236	CON-PV_15_12108	Wild-type	CD	3 days	Control	Portal Vein Serum	15	Water
SA207237	CON-PV_30_12118	Wild-type	CD	3 days	Control	Portal Vein Serum	30	Water
SA207238	CON-PV_30_12116	Wild-type	CD	3 days	Control	Portal Vein Serum	30	Water
SA207239	CON-PV_30_12117	Wild-type	CD	3 days	Control	Portal Vein Serum	30	Water
SA207240	CON-PV_30_12119	Wild-type	CD	3 days	Control	Portal Vein Serum	30	Water
SA207241	CON-PV_30_12120	Wild-type	CD	3 days	Control	Portal Vein Serum	30	Water
SA207242	CON-PV_45_12126	Wild-type	CD	3 days	Control	Portal Vein Serum	45	Water
SA207243	CON-PV_45_12127	Wild-type	CD	3 days	Control	Portal Vein Serum	45	Water
SA207244	CON-PV_45_12128	Wild-type	CD	3 days	Control	Portal Vein Serum	45	Water
SA207245	CON-PV_45_12129	Wild-type	CD	3 days	Control	Portal Vein Serum	45	Water
SA207246	CON-PV_45_12130	Wild-type	CD	3 days	Control	Portal Vein Serum	45	Water
SA207247	CON-PV_60_12136	Wild-type	CD	3 days	Control	Portal Vein Serum	60	Water
SA207248	CON-PV_60_12139	Wild-type	CD	3 days	Control	Portal Vein Serum	60	Water
SA207249	CON-PV_60_12137	Wild-type	CD	3 days	Control	Portal Vein Serum	60	Water
SA207250	CON-PV_60_12140	Wild-type	CD	3 days	Control	Portal Vein Serum	60	Water
SA207251	CON-PV_60_12138	Wild-type	CD	3 days	Control	Portal Vein Serum	60	Water
SA207252	CON-PV_90_12146	Wild-type	CD	3 days	Control	Portal Vein Serum	90	Water
SA207253	CON-PV_90_12148	Wild-type	CD	3 days	Control	Portal Vein Serum	90	Water
SA207254	CON-PV_90_12147	Wild-type	CD	3 days	Control	Portal Vein Serum	90	Water
SA207221	CON-PV_0_13158	Wild-type	CD	3 days	Control	Portal Vein Serum	-	Water
SA207222	CON-PV_0_13157	Wild-type	CD	3 days	Control	Portal Vein Serum	-	Water
SA207223	CON-PV_0_13156	Wild-type	CD	3 days	Control	Portal Vein Serum	-	Water
SA207261	13C-Fruc_LFD-HV_13171	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207262	13C-Fruc_LFD-HV_13170	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207263	13C-Fruc_LFD-HV_13169	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207264	KHK_Gly_6_010	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207265	KHK_Gly_3_010	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207266	KHK_Gly_2_010	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207267	13C-Fruc_LFD-HV_13168	Wild-type	CD	3 days	Control	Systemic Serum	10	Water
SA207268	13C-Fruc_LFD-HV_13172	Wild-type	CD	3 days	Control	Systemic Serum	10	Water

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Collection:

Collection ID:	CO002240
Collection Summary:	After 3 days or 4 weeks of either HFD or CD feeding, each mouse was administrated a 1g/kg U-13C fructose oral gavage. Mice were anesthetized with 2% isoflurane, and their abdominal cavities were opened at the specified time. Abdominal tissues were displaced to identify the portal vein, and a 27G needle was used to immediately collect portal vein blood samples (~100 µL). Immediately after, cardiac blood was collected from the right ventricle using a 25G needle (~1 mL). The jejunum, pancreas, and liver tissues were quickly resected and snap-frozen in liquid nitrogen. Blood samples were placed on ice in the absence of anticoagulant for 20 minutes and centrifuged at 16,000 × g for 10 minutes at 4 °C to separate the serum and plasma fractions of each sample. Serum and tissue samples were kept at −80 °C until further analysis.
Collection Protocol Filename:	cwwong_methods.pdf
Sample Type:	Tissue

Treatment:

Treatment ID:	TR002259
Treatment Summary:	After 3 days or 4 weeks of either HFD or CD feeding, each mouse was administrated a 1g/kg U-13C fructose oral gavage. For ad-lib fructose feeding experiments, mice were fed fructose in the drinking water (15% w/v) for 4 weeks. Saline, fructose and glycerate injection were done in CD-fed mice for 12 weeks. For glycerate kinetic study, glycerate injections were performed in treatment-navie mice.

Sample Preparation:

Sampleprep ID:	SP002253
Sampleprep Summary:	For LC-MS: Metabolites were extracted using a protocol optimized for water-soluble polar metabolite analysis using liquid chromatography coupled with mass spectrometry. All extraction buffers were stored at -20°C prior to usage and immediately preceding the metabolite extraction. For serum samples, a 10 μL serum aliquot was used for fructose tracing. Metabolites were extracted with 40 μL of ice-cold methanol and incubated at -20 °C for 20 minutes. The clean supernatant was collected after centrifugation for 10 minutes at the highest speed, and the leftover pellet was further treated with 200 µL cold extraction buffer (40:40:20 v/v/v methanol:acetonitrile:water solution) and left to incubate on crushed ice for an additional 10 minutes. Following an additional 10 minutes of centrifugation at the highest speed, the clean supernatant was collected and pooled with the supernatant from the first collection. For tissue samples, the extraction buffer used was a (v/v/v) solution of 40:40:20 (methanol:acetonitrile:water) + 0.1 M formic acid. An aliquot volume equivalent to 20x the sample weight was added to the Eppendorf tube with the homogenized sample, vortexed for 10 seconds, and left to chill on crushed ice for 10 minutes. The samples were then centrifuged for 10 minutes at the highest speed at 4°C. The supernatant was transferred to a correspondingly labeled and chilled Eppendorf tube, and the process was repeated once more. The total volume of supernatant was then centrifuged for an additional 10 minutes. After centrifugation, a final 500µL aliquot of the homogenate was then pipetted to a second clean Eppendorf tube, to which 44µL of 15% (m/v) NH4HCO3 was added to neutralize the acid in the buffer and precipitate the protein. This is the final sample extract to be vialed and loaded to the instrument for analysis. Metabolite extracts were stored at -80 °C until analysis. For MSI: Pancreas tissues were first equilibrated to -15°C then sectioned to 20 µm thickness using a Leica CM1950 cryostat (Buffalo Grove, IL, USA). Cut sections were then thaw-mounted on clean microscope slides (1 mm height, plain, Fisher Scientific, Pittsburgh, PA) and stored at -80 °C until IR-MALDESI-MSI analysis. Tissues were first blocked into four groups, each containing one replicate of each condition. Tissues were both cut and imaged in randomized order within these blocks to minimize sampling bias.

Sampleprep ID:

SP002253

Sampleprep Summary:

For LC-MS: Metabolites were extracted using a protocol optimized for water-soluble polar metabolite analysis using liquid chromatography coupled with mass spectrometry. All extraction buffers were stored at -20°C prior to usage and immediately preceding the metabolite extraction. For serum samples, a 10 μL serum aliquot was used for fructose tracing. Metabolites were extracted with 40 μL of ice-cold methanol and incubated at -20 °C for 20 minutes. The clean supernatant was collected after centrifugation for 10 minutes at the highest speed, and the leftover pellet was further treated with 200 µL cold extraction buffer (40:40:20 v/v/v methanol:acetonitrile:water solution) and left to incubate on crushed ice for an additional 10 minutes. Following an additional 10 minutes of centrifugation at the highest speed, the clean supernatant was collected and pooled with the supernatant from the first collection. For tissue samples, the extraction buffer used was a (v/v/v) solution of 40:40:20 (methanol:acetonitrile:water) + 0.1 M formic acid. An aliquot volume equivalent to 20x the sample weight was added to the Eppendorf tube with the homogenized sample, vortexed for 10 seconds, and left to chill on crushed ice for 10 minutes. The samples were then centrifuged for 10 minutes at the highest speed at 4°C. The supernatant was transferred to a correspondingly labeled and chilled Eppendorf tube, and the process was repeated once more. The total volume of supernatant was then centrifuged for an additional 10 minutes. After centrifugation, a final 500µL aliquot of the homogenate was then pipetted to a second clean Eppendorf tube, to which 44µL of 15% (m/v) NH4HCO3 was added to neutralize the acid in the buffer and precipitate the protein. This is the final sample extract to be vialed and loaded to the instrument for analysis. Metabolite extracts were stored at -80 °C until analysis. For MSI: Pancreas tissues were first equilibrated to -15°C then sectioned to 20 µm thickness using a Leica CM1950 cryostat (Buffalo Grove, IL, USA). Cut sections were then thaw-mounted on clean microscope slides (1 mm height, plain, Fisher Scientific, Pittsburgh, PA) and stored at -80 °C until IR-MALDESI-MSI analysis. Tissues were first blocked into four groups, each containing one replicate of each condition. Tissues were both cut and imaged in randomized order within these blocks to minimize sampling bias.

Combined analysis:

Analysis ID	AN003540	AN003541
Analysis type	MS	MS
Chromatography type	HILIC	IR-MALDI
Chromatography system	Thermo Vanquish	Other
Column	Waters XBridge BEH Amide (150mm x 2.1mm,2.5um)	Other
MS Type	ESI	ESI
MS instrument type	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive Plus Orbitrap	Thermo Exploris 240 Orbitrap
Ion Mode	NEGATIVE	NEGATIVE
Units	Total Abundance	Peak Area

Chromatography:

Chromatography ID:	CH002614
Instrument Name:	Thermo Vanquish
Column Name:	Waters XBridge BEH Amide (150mm x 2.1mm,2.5um)
Chromatography Type:	HILIC

Chromatography ID:	CH002615
Instrument Name:	Other
Column Name:	Other
Chromatography Type:	IR-MALDI

MS:

MS ID:	MS003298
Analysis ID:	AN003540
Instrument Name:	Thermo Q Exactive Plus Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	Fullscan mass spectrometry. The full scan mass spectrometry analysis was performed on a Thermo Q Exactive PLUS with a HESI source which was set to a spray voltage of -2.7kV under negative mode and 3.5kV under positive mode. The sheath, auxiliary, and sweep gas flow rates of 40, 10, and 2 (arbitrary unit), respectively. The capillary temperature was set to 300°C, and the aux gas heater was 360°C. The S-lens RF level was 45. The m/z range was set to 72 to 1000 m/z under both positive and negative ionization mode. The AGC target was set to 3e6, and the maximum IT was 200 ms. The mass resolution (full-width half maximum) was set to 70,000 @ m/z = 200.
Ion Mode:	NEGATIVE
Analysis Protocol File:	cwwong_methods.pdf

MS ID:	MS003299
Analysis ID:	AN003541
Instrument Name:	Thermo Exploris 240 Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	To ablate targeted tissue regions a 2970-nm wavelength laser was used with a single burst of ten pulses to produce 1 mJ of energy at a rate of 10 kHz. X and Y stage movements of 100 µm were used to achieve oversampling(Nazari and Muddiman, 2015). Ablated analytes were post-ionized in the orthogonal electrospray plume, established by applying a voltage of approximately 3 kV to the electrospray solvent (1 mM acetic acid in 50:50 water/acetonitrile.) Mass spectrometry analysis of ionized molecules was performed in negative mode with internal calibrant used to achieve high mass accuracy (<2.5 ppm) within the 85-225 m/z range. Automatic gain control (AGC) was disabled. A mass resolution power of 240,000FWHM at 200 m/z was used with a fixed injection time (15 ms) to synchronize timing of the ablation plume with ion collection in the C-trap of the mass spectrometer.
Ion Mode:	NEGATIVE
Analysis Protocol File:	cwwong_methods.pdf