Summary of Study ST003812

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 PR002385. The data can be accessed directly via it's Project DOI: 10.21228/M8BR77 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	ST003812
Study Title	Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle - metabolomics from CD36 x PdgfBcre vs WT mice on control or SAAR diet
Study Summary	Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle β-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. We performed metabolomics analysis on the extensor digitorum longus (EDL), soleus (Sol), and serum to investigate the systemic effects of genotype and diet on muscle and circulatory metabolites. After one week of dietary treatment, significant changes in a wide range of metabolites were not observed. Interestingly, only minimal differences were detected between the WT and CD36 × PDGFb KO models.
Institute	Princeton University
Department	Department of Chemistry
Last Name	Mann
First Name	Charlotte Greta
Address	Joseph Stelzmann Strasse 26, 50931 Cologne, Germany
Email	cmann5@uni-koeln.de
Phone	+49 221 478-84102
Submit Date	2025-03-19
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2025-04-18
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR002385
Project DOI:	doi: 10.21228/M8BR77
Project Title:	Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle
Project Summary:	Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle β-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability.
Institute:	Princeton University
Department:	Department of Chemistry
Laboratory:	Prof. Dr. Joshua Rabinowitz
Last Name:	Mann
First Name:	Charlotte G.
Address:	Joseph Stelzmann Strasse 26, 50931 Cologne, Germany
Email:	cmann5@uni-koeln.de
Phone:	+49 221 478-84102

Subject:

Subject ID:	SU003946
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	CD36 fl/fl x Pdgfb-Cre
Age Or Age Range:	14 weeks
Gender:	Male

Factors:

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

mb_sample_id	local_sample_id	Sample source	Genoytype	Diet
SA417473	b_3	-	-	-
SA417474	b_2	-	-	-
SA417475	b_1	-	-	-
SA417476	Sa_8	Mouse EDL muscle	Cre hetero	CON
SA417477	Sa_9	Mouse EDL muscle	Cre hetero	CON
SA417478	Sa_25	Mouse EDL muscle	Cre hetero	CON
SA417479	Sa_23	Mouse EDL muscle	Cre hetero	CON
SA417480	Sa_19	Mouse EDL muscle	Cre hetero	CON
SA417481	Sa_17	Mouse EDL muscle	Cre hetero	CON
SA417482	Sa_16	Mouse EDL muscle	Cre hetero	SAAR
SA417483	Sa_2	Mouse EDL muscle	Cre hetero	SAAR
SA417484	Sa_26	Mouse EDL muscle	Cre hetero	SAAR
SA417485	Sa_24	Mouse EDL muscle	Cre hetero	SAAR
SA417486	Sa_1	Mouse EDL muscle	Cre hetero	SAAR
SA417487	Sa_5	Mouse EDL muscle	Cre hetero	SAAR
SA417488	Sa_13	Mouse EDL muscle	Cre hetero	SAAR
SA417489	Sa_7	Mouse EDL muscle	wild type	Con
SA417490	Sa_6	Mouse EDL muscle	wild type	Con
SA417491	Sa_20	Mouse EDL muscle	wild type	Con
SA417492	Sa_21	Mouse EDL muscle	wild type	Con
SA417493	Sa_4	Mouse EDL muscle	wild type	Con
SA417494	Sa_3	Mouse EDL muscle	wild type	Con
SA417495	Sa_15	Mouse EDL muscle	wild type	SAAR
SA417496	Sa_14	Mouse EDL muscle	wild type	SAAR
SA417497	Sa_10	Mouse EDL muscle	wild type	SAAR
SA417498	Sa_11	Mouse EDL muscle	wild type	SAAR
SA417499	Sa_12	Mouse EDL muscle	wild type	SAAR
SA417500	Sa_22	Mouse EDL muscle	wild type	SAAR
SA417501	Sa_18	Mouse EDL muscle	wild type	SAAR
SA417502	Sa_60	Mouse Serum	Cre hetero	CON
SA417503	Sa_69	Mouse Serum	Cre hetero	CON
SA417504	Sa_71	Mouse Serum	Cre hetero	CON
SA417505	Sa_77	Mouse Serum	Cre hetero	CON
SA417506	Sa_75	Mouse Serum	Cre hetero	CON
SA417507	Sa_61	Mouse Serum	Cre hetero	CON
SA417508	Sa_54	Mouse Serum	Cre hetero	SAAR
SA417509	Sa_53	Mouse Serum	Cre hetero	SAAR
SA417510	Sa_76	Mouse Serum	Cre hetero	SAAR
SA417511	Sa_68	Mouse Serum	Cre hetero	SAAR
SA417512	Sa_65	Mouse Serum	Cre hetero	SAAR
SA417513	Sa_78	Mouse Serum	Cre hetero	SAAR
SA417514	Sa_57	Mouse Serum	Cre hetero	SAAR
SA417515	Sa_55	Mouse Serum	wild type	Con
SA417516	Sa_58	Mouse Serum	wild type	Con
SA417517	Sa_59	Mouse Serum	wild type	Con
SA417518	Sa_72	Mouse Serum	wild type	Con
SA417519	Sa_56	Mouse Serum	wild type	Con
SA417520	Sa_73	Mouse Serum	wild type	Con
SA417521	Sa_62	Mouse Serum	wild type	SAAR
SA417522	Sa_63	Mouse Serum	wild type	SAAR
SA417523	Sa_64	Mouse Serum	wild type	SAAR
SA417524	Sa_66	Mouse Serum	wild type	SAAR
SA417525	Sa_67	Mouse Serum	wild type	SAAR
SA417526	Sa_70	Mouse Serum	wild type	SAAR
SA417527	Sa_74	Mouse Serum	wild type	SAAR
SA417528	Sa_49	Mouse soleus muscle	Cre hetero	CON
SA417529	Sa_45	Mouse soleus muscle	Cre hetero	CON
SA417530	Sa_34	Mouse soleus muscle	Cre hetero	CON
SA417531	Sa_35	Mouse soleus muscle	Cre hetero	CON
SA417532	Sa_43	Mouse soleus muscle	Cre hetero	CON
SA417533	Sa_51	Mouse soleus muscle	Cre hetero	CON
SA417534	Sa_52	Mouse soleus muscle	Cre hetero	SAAR
SA417535	Sa_39	Mouse soleus muscle	Cre hetero	SAAR
SA417536	Sa_27	Mouse soleus muscle	Cre hetero	SAAR
SA417537	Sa_28	Mouse soleus muscle	Cre hetero	SAAR
SA417538	Sa_31	Mouse soleus muscle	Cre hetero	SAAR
SA417539	Sa_42	Mouse soleus muscle	Cre hetero	SAAR
SA417540	Sa_50	Mouse soleus muscle	Cre hetero	SAAR
SA417541	Sa_33	Mouse soleus muscle	wild type	Con
SA417542	Sa_46	Mouse soleus muscle	wild type	Con
SA417543	Sa_32	Mouse soleus muscle	wild type	Con
SA417544	Sa_47	Mouse soleus muscle	wild type	Con
SA417545	Sa_30	Mouse soleus muscle	wild type	Con
SA417546	Sa_29	Mouse soleus muscle	wild type	Con
SA417547	Sa_40	Mouse soleus muscle	wild type	SAAR
SA417548	Sa_41	Mouse soleus muscle	wild type	SAAR
SA417549	Sa_37	Mouse soleus muscle	wild type	SAAR
SA417550	Sa_36	Mouse soleus muscle	wild type	SAAR
SA417551	Sa_44	Mouse soleus muscle	wild type	SAAR
SA417552	Sa_48	Mouse soleus muscle	wild type	SAAR
SA417553	Sa_38	Mouse soleus muscle	wild type	SAAR

Showing results 1 to 81 of 81

Showing results 1 to 81 of 81

Collection:

Collection ID:	CO003939
Collection Summary:	Tissues were collected and snap-frozen in liquid nitrogen. Blood was collected by cardiac puncture, immediately placed on ice, centrifuged, and the serum was collected, aliquoted, and frozen in liquid nitrogen.
Sample Type:	muscle and serum

Treatment:

Treatment ID:	TR003955
Treatment Summary:	Wild type (WT) C57BL/6J and Cd36 LoxP/LoxP mice (Cd36tm1.1Ijg/J) were purchased from Charles River (Freiburg im Breisgau, Germany). To obtain inducible endothelial cell-specific Cd36 knockout (ECCD36-/-) mice, Cd36 LoxP/LoxP mice were crossed with PDGFβ.iCreER mice, an EC-selective inducible Cre-driver line (Claxton et al., 2008, DOI: 10.1002/dvg.20367). Recombination was induced in 8-10 weeks old male mice by daily intraperitoneal (i.p.) administration of 1mg tamoxifen (T5648, Sigma-Aldrich) dissolved in 1:10 ethanol:corn oil solution for 3 consecutive days. A wash out period of at least seven days was allowed before starting the experiments. Tamoxifen-treated Cre-negative littermates were used as control for all experiments. Experimental diets were based on Research Diets D12450J with approximately 18% of calories from protein, 10% from fat and 72% from carbohydrates. SAAR diets containing 1.15g methionine (M)/kg food and lacking cysteine (C) (Miller et al., 2005, DOI: 10.1111/j.1474-9726.2005.00152.x) in the context of a 17% protein/ 73% carbohydrate calorie diet were provided Ad Libitum (AL). Food intake was monitored daily during experiments. The Research Diets product number for the control diet is A17101101 and for SAAR diet is A17101103.

Treatment ID:

TR003955

Treatment Summary:

Wild type (WT) C57BL/6J and Cd36 LoxP/LoxP mice (Cd36tm1.1Ijg/J) were purchased from Charles River (Freiburg im Breisgau, Germany). To obtain inducible endothelial cell-specific Cd36 knockout (ECCD36-/-) mice, Cd36 LoxP/LoxP mice were crossed with PDGFβ.iCreER mice, an EC-selective inducible Cre-driver line (Claxton et al., 2008, DOI: 10.1002/dvg.20367). Recombination was induced in 8-10 weeks old male mice by daily intraperitoneal (i.p.) administration of 1mg tamoxifen (T5648, Sigma-Aldrich) dissolved in 1:10 ethanol:corn oil solution for 3 consecutive days. A wash out period of at least seven days was allowed before starting the experiments. Tamoxifen-treated Cre-negative littermates were used as control for all experiments. Experimental diets were based on Research Diets D12450J with approximately 18% of calories from protein, 10% from fat and 72% from carbohydrates. SAAR diets containing 1.15g methionine (M)/kg food and lacking cysteine (C) (Miller et al., 2005, DOI: 10.1111/j.1474-9726.2005.00152.x) in the context of a 17% protein/ 73% carbohydrate calorie diet were provided Ad Libitum (AL). Food intake was monitored daily during experiments. The Research Diets product number for the control diet is A17101101 and for SAAR diet is A17101103.

Sample Preparation:

Sampleprep ID:	SP003952
Sampleprep Summary:	Metabolite extraction of serum: Serum (3 μl) was extracted with cold 100% methanol (40X), vortexed, and incubated on dry ice for 30 min. Then, the extract was centrifuged at 20,000 x g for 20 minutes at 4°C and supernatant was transferred to tubes containing 100% methanol, vortexed and incubated on dry ice for 30 min. Then, the extract was centrifuged at 20,000 x g for 20 minutes at 4°C and supernatant was transferred to tubes for LC-MS analysis. Metabolite extraction of tissues: Frozen tissue pieces were pulverized using a Cryomill (Retsch) at cryogenic temperature. Ground tissue was weighed (10–20 mg) and transferred into a precooled tube for extraction. Soluble metabolites extraction was done by adding −20 °C 40:40:20 methanol:acetonitrile:water to the resulting powder (40 μl solvent per mg tissue). Samples were vortexed for 10 seconds, cooled at 4°C (on wet ice) for 20 minutes and then centrifuged at 4 °C at 20,000 x g for 30 minutes. Supernatant was transferred to LC–MS vials for analysis.

Sampleprep ID:

SP003952

Sampleprep Summary:

Metabolite extraction of serum: Serum (3 μl) was extracted with cold 100% methanol (40X), vortexed, and incubated on dry ice for 30 min. Then, the extract was centrifuged at 20,000 x g for 20 minutes at 4°C and supernatant was transferred to tubes containing 100% methanol, vortexed and incubated on dry ice for 30 min. Then, the extract was centrifuged at 20,000 x g for 20 minutes at 4°C and supernatant was transferred to tubes for LC-MS analysis. Metabolite extraction of tissues: Frozen tissue pieces were pulverized using a Cryomill (Retsch) at cryogenic temperature. Ground tissue was weighed (10–20 mg) and transferred into a precooled tube for extraction. Soluble metabolites extraction was done by adding −20 °C 40:40:20 methanol:acetonitrile:water to the resulting powder (40 μl solvent per mg tissue). Samples were vortexed for 10 seconds, cooled at 4°C (on wet ice) for 20 minutes and then centrifuged at 4 °C at 20,000 x g for 30 minutes. Supernatant was transferred to LC–MS vials for analysis.

Combined analysis:

Analysis ID	AN006267	AN006268
Analysis type	MS	MS
Chromatography type	HILIC	HILIC
Chromatography system	Thermo Dionex	Thermo Dionex
Column	Waters XBridge BEH Amide (100 x 2.1mm,2.5um)	Waters XBridge BEH Amide (100 x 2.1mm,2.5um)
MS Type	ESI	ESI
MS instrument type	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive Plus Orbitrap	Thermo Orbitrap Exploris 240
Ion Mode	POSITIVE	NEGATIVE
Units	ion count	ion count

Chromatography:

Chromatography ID:	CH004755
Instrument Name:	Thermo Dionex
Column Name:	Waters XBridge BEH Amide (100 x 2.1mm,2.5um)
Column Temperature:	25°C
Flow Gradient:	0 minutes, 85% B; 2 minutes, 85% B; 3 minutes, 80% B; 5 minutes, 80% B; 6 minutes, 75% B; 7 minutes, 75% B; 8 minutes, 70% B; 9 minutes, 70% B; 10 minutes, 50% B; 12 minutes, 50% B; 13 minutes, 25% B; 16 minutes, 25% B; 18 minutes, 0% B; 23 minutes, 0% B; 24 minutes, 85% B; 30 minutes, 85% B, gradient changes were linear
Flow Rate:	150 μl min−1
Solvent A:	95% water/5% acetonitrile; 20 mM ammonium acetate; 20 mM ammonium hydroxide, pH 9.4
Solvent B:	100% acetonitrile
Chromatography Type:	HILIC

MS:

MS ID:	MS005969
Analysis ID:	AN006267
Instrument Name:	Thermo Q Exactive Plus Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	MS full scans were in negative or positive ion mode with a resolution of 140,000 at m/z 200 and scan range of 70–1,000 m/z. The automatic gain control (AGC) target was 1 × 106. LC-MS peak files were analyzed and visualized with El-MAVEN (Elucidata) using 5 ppm ion extraction window, minimum peak intensity of 1 x 105 ions, and minimum signal to background blank ratio of 2
Ion Mode:	POSITIVE

MS ID:	MS005970
Analysis ID:	AN006268
Instrument Name:	Thermo Orbitrap Exploris 240
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	MS full scans were in negative or positive ion mode with a resolution of 140,000 at m/z 200 and scan range of 70–1,000 m/z. The automatic gain control (AGC) target was 1 × 106. LC-MS peak files were analyzed and visualized with El-MAVEN
Ion Mode:	NEGATIVE