Summary of Study ST004278

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 PR001984. The data can be accessed directly via it's Project DOI: 10.21228/M88J0W 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	ST004278
Study Title	Investigating the immediate metabolic effects of acute alpha-ketoglutarate supplementation in mice
Study Summary	Alpha-ketoglutarate (aKG) supplementation extends healthspan and lifespan in mice and several other model organisms, and our metabolic atlas of mouse aging showed that plasma aKG levels drop with age. The immediate metabolic impacts of aKG supplementation are not known. In this study, we treated young and old male mice with aKG and measured metabolite levels in heart, kidney, liver, lung, quad muscle and spleen after 3 hours. Metabolic changes were limited suggesting that aKG exerts its positive effects via long term metabolic and cellular rewiring.
Institute	University of Southern California
Department	Keck School of Medicine
Laboratory	Mullen Lab
Last Name	Pilley
First Name	Steven
Address	1501 San Pablo St., Los Angeles, CA, 90033, USA
Email	pilley@usc.edu
Phone	+13234427555
Submit Date	2025-08-19
Num Groups	4
Total Subjects	19
Num Males	19
Raw Data Available	Yes
Raw Data File Type(s)	mzML
Analysis Type Detail	LC-MS
Release Date	2025-11-19
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001984
Project DOI:	doi: 10.21228/M88J0W
Project Title:	A metabolic atlas of mouse aging
Project Summary:	Humans are living longer, but this is accompanied by an increased incidence of age-related chronic diseases. Many of these diseases are influenced by age-associated metabolic dysregulation, but how metabolism changes in multiple organs during aging in males and females is not known. Answering this could reveal new mechanisms of aging and age-targeted therapeutics. In this study, we describe how metabolism changes in 12 organs in male and female mice at 5 different ages. Organs show distinct patterns of metabolic aging that are affected by sex differently. Hydroxyproline shows the most consistent change across the dataset, decreasing with age in 11 out of 12 organs investigated. We also developed a metabolic aging clock that predicts biological age and identified alpha-ketoglutarate, previously shown to extend lifespan in mice, as a key predictor of age. Our results reveal fundamental insights into the aging process and identify new therapeutic targets to maintain organ health.
Institute:	University of Southern California
Department:	Keck School of Medicine
Laboratory:	Mullen Lab
Last Name:	Pilley
First Name:	Steven
Address:	1501 San Pablo St., Los Angeles, CA, 90033, USA
Email:	pilley@usc.edu
Phone:	+13234427555
Funding Source:	Impetus Grants 014746-00001

Subject:

Subject ID:	SU004431
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6J
Age Or Age Range:	3-18 months
Gender:	Male
Animal Animal Supplier:	Jackson Lab

Factors:

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

mb_sample_id	local_sample_id	Injection order	Sample source	Age	Treatment
SA498117	80	100	Kidney	COM	PBS
SA498118	86	101	Kidney	CYM	PBS
SA498119	89	102	Kidney	CYM	PBS
SA498120	83	103	Kidney	COM	AKG
SA498121	79	104	Kidney	COM	PBS
SA498122	Kidney pool 2	105	Pool	N/A	N/A
SA498123	81	106	Kidney	COM	AKG
SA498124	85	107	Kidney	COM	AKG
SA498125	87	108	Kidney	CYM	PBS
SA498126	95	109	Kidney	CYM	AKG
SA498127	18	10	Liver	CYM	AKG
SA498128	94	110	Kidney	CYM	AKG
SA498129	93	111	Kidney	CYM	AKG
SA498130	88	112	Kidney	CYM	PBS
SA498131	78	113	Kidney	COM	PBS
SA498132	82	114	Kidney	COM	AKG
SA498133	Kidney pool 3	115	Pool	N/A	N/A
SA498134	Master pool 6	116	Pool	N/A	N/A
SA498135	Heart pool 1	117	Pool	N/A	N/A
SA498136	96	118	Heart	COM	PBS
SA498137	114	119	Heart	CYM	AKG
SA498138	7	11	Liver	COM	AKG
SA498139	110	120	Heart	CYM	AKG
SA498140	101	121	Heart	COM	AKG
SA498141	105	122	Heart	CYM	PBS
SA498142	111	123	Heart	CYM	AKG
SA498143	103	124	Heart	COM	AKG
SA498144	108	125	Heart	CYM	PBS
SA498145	102	126	Heart	COM	AKG
SA498146	Heart pool 3	127	Pool	N/A	N/A
SA498147	97	128	Heart	COM	PBS
SA498148	Heart pool 2	129	Pool	N/A	N/A
SA498149	16	12	Liver	CYM	AKG
SA498150	107	130	Heart	CYM	PBS
SA498151	104	131	Heart	COM	AKG
SA498152	99	132	Heart	COM	PBS
SA498153	112	133	Heart	CYM	AKG
SA498154	109	134	Heart	CYM	PBS
SA498155	98	135	Heart	COM	PBS
SA498156	106	136	Heart	CYM	PBS
SA498157	14	13	Liver	CYM	PBS
SA498158	15	14	Liver	CYM	AKG
SA498159	Liver pool 2	15	Pool	N/A	N/A
SA498160	8	16	Liver	COM	AKG
SA498161	5	17	Liver	COM	AKG
SA498162	1	18	Liver	COM	PBS
SA498163	11	19	Liver	CYM	PBS
SA498164	Blank 1	1	Blank	N/A	N/A
SA498165	17	20	Liver	CYM	AKG
SA498166	4	21	Liver	COM	PBS
SA498167	12	22	Liver	CYM	PBS
SA498168	19	23	Liver	CYM	AKG
SA498169	3	24	Liver	COM	PBS
SA498170	Liver pool 3	25	Pool	N/A	N/A
SA498171	Master pool 2	26	Pool	N/A	N/A
SA498172	Lung pool 1	27	Pool	N/A	N/A
SA498173	24	28	Lung	COM	AKG
SA498174	30	29	Lung	CYM	PBS
SA498175	Master pool 1	2	Pool	N/A	N/A
SA498176	31	30	Lung	CYM	PBS
SA498177	34	31	Lung	CYM	AKG
SA498178	28	32	Lung	COM	AKG
SA498179	22	33	Lung	COM	PBS
SA498180	27	34	Lung	COM	AKG
SA498181	20	35	Lung	COM	PBS
SA498182	36	36	Lung	CYM	AKG
SA498183	29	37	Lung	CYM	PBS
SA498184	Lung pool 2	38	Pool	N/A	N/A
SA498185	Spleen pool 1	39	Pool	N/A	N/A
SA498186	CellQC 1	3	QC	N/A	N/A
SA498187	25	40	Lung	COM	AKG
SA498188	38	41	Lung	CYM	AKG
SA498189	37	42	Lung	CYM	AKG
SA498190	26	43	Lung	COM	AKG
SA498191	33	44	Lung	CYM	PBS
SA498192	32	45	Lung	CYM	PBS
SA498193	21	46	Lung	COM	PBS
SA498194	35	47	Lung	CYM	AKG
SA498195	23	48	Lung	COM	PBS
SA498196	Lung pool 3	49	Pool	N/A	N/A
SA498197	Liver pool 1	4	Pool	N/A	N/A
SA498198	Master pool 3	50	Pool	N/A	N/A
SA498199	Spleen pool 2	51	Pool	N/A	N/A
SA498200	40	52	Spleen	COM	PBS
SA498201	41	53	Spleen	COM	PBS
SA498202	47	54	Spleen	COM	AKG
SA498203	48	55	Spleen	CYM	PBS
SA498204	54	56	Spleen	CYM	AKG
SA498205	51	57	Spleen	CYM	PBS
SA498206	42	58	Spleen	COM	PBS
SA498207	57	59	Spleen	CYM	AKG
SA498208	2	5	Liver	COM	PBS
SA498209	Spleen pool 3	60	Pool	N/A	N/A
SA498210	43	61	Spleen	COM	AKG
SA498211	45	62	Spleen	COM	AKG
SA498212	55	63	Spleen	CYM	AKG
SA498213	49	64	Spleen	CYM	PBS
SA498214	39	65	Spleen	COM	PBS
SA498215	56	66	Spleen	CYM	AKG
SA498216	50	67	Spleen	CYM	PBS

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Collection:

Collection ID:	CO004424
Collection Summary:	We humanely euthanized mice 3 hours after intraperitoneal injection of PBS or AKG, and flash-froze tissues.
Sample Type:	Liver, Lung, Quadriceps muscle, Kidney, Spleen, Heart

Treatment:

Treatment ID:	TR004440
Treatment Summary:	We acquired 10 old (17-months-old) and 10 young (2-months-old) male C57Bl/6J mice from Jackson Lab and allowed them to habituate for 1 month before randomly assigning them equally into control or treatment groups. We intraperitoneally injected the control mice with PBS and the treatment mice with alpha-ketoglutarate (aKG, Sigma Aldrich, 75890) at 10 mg/kg body weight. There were 4 groups overall: 1) old control (COM_PBS); 2) old aKG (COM_aKG); 3) young control (CYM_PBS); and 4) young aKG (CYM_aKG). All groups n = 5, except for old control where n = 4.

Treatment ID:

TR004440

Treatment Summary:

We acquired 10 old (17-months-old) and 10 young (2-months-old) male C57Bl/6J mice from Jackson Lab and allowed them to habituate for 1 month before randomly assigning them equally into control or treatment groups. We intraperitoneally injected the control mice with PBS and the treatment mice with alpha-ketoglutarate (aKG, Sigma Aldrich, 75890) at 10 mg/kg body weight. There were 4 groups overall: 1) old control (COM_PBS); 2) old aKG (COM_aKG); 3) young control (CYM_PBS); and 4) young aKG (CYM_aKG). All groups n = 5, except for old control where n = 4.

Sample Preparation:

Sampleprep ID:	SP004437
Sampleprep Summary:	We homogenized samples in 0.3% formic acid in 1:1 Acetonitrile/Water and dried samples. We reconstituted dried supernatants in 50 uL of LC-grade Methanol: Ultrapure-water mix (1:1) .

Chromatography:

Chromatography ID:	CH005407
Instrument Name:	Agilent 1290 Infinity II
Column Name:	Agilent ZORBAX RRHD SB-C18 (150 x 2.1mm,1.8um)
Column Temperature:	35
Flow Gradient:	0.25 mL/min, 0–2.5 min, 100% A; 7.5 min, 80% A and 20% B; 13 min 55% A and 45% B; 20 min, 1% A and 99% B; 24 min, 1% A and 99% B; 24.05 min, 1% A and 99% C; 27 min, 1% A and 99% C; at 0.8 mL/min, 27.5–31.35 min, 1% A and 99% C; at 0.6 mL/min, 31.50 min, 1% A and 99% C; at 0.4 mL/min, 32.25–39.9 min, 100% A; and at 0.25 mL/min, 40 min, 100% A.
Flow Rate:	0.25-0.8 mL/min
Solvent A:	97% water/3% methanol;15 mM acetic acid; 10 mM tributylamine at pH 5
Solvent B:	100% methanol; 15 mM acetic acid; 10 mM tributylamine
Chromatography Type:	Reversed phase
Solvent C:	100% acetonitrile, wash solvent.

Analysis:

Analysis ID:	AN007117
Laboratory Name:	Lyssiotis Lab
Analysis Type:	MS
Operator Name:	Li Zhang
Chromatography ID:	CH005407
Num Factors:	136
Num Metabolites:	117
Units:	AU