Summary of Study ST001507

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 PR001017. The data can be accessed directly via it's Project DOI: 10.21228/M87M4S 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	ST001507
Study Title	Hepatic [U-13C]Lactate tracing and metabolomics in young and old WT and SIRT6 overexpressing mice
Study Summary	Our previous data had suggested that gluconeogenesis capacity from the precursors lactate and glycerol declines in old C57BL/6 mice. This decline was rescued by whole body SIRT6 overexpression. Since the liver is the main gluconeogenic organ, we performed here liver metabolomics in young (6 months) versus old (20-24 months) WT and SIRT6-transgenic mice. We used liver tissues of 6h morning-fasted mice, either in naïve state or 15 minutes after intraperitoneal injection of 1mg/kg [U-13C]Lactate.
Institute	Bar Ilan University
Department	The Mina & Everard Goodman Faculty of Life Sciences
Laboratory	Laboratory of Prof. Haim Cohen
Last Name	Cohen
First Name	Chaim
Address	Bar Ilan University, Ramat Gan, Israel, 5290002
Email	Haim.Cohen@biu.ac.il
Phone	+97235318383
Submit Date	2020-10-06
Num Groups	8
Total Subjects	40
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2021-03-11
Release Version	1

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Project:

Project ID:	PR001017
Project DOI:	doi: 10.21228/M87M4S
Project Title:	Hepatic [U-13C]Lactate tracing and metabolomics in young and old WT and SIRT6 overexpressing mice
Project Type:	MS analysis
Project Summary:	Our previous data had suggested that gluconeogenesis capacity from the precursors lactate and glycerol declines in old C57BL/6 mice. This decline was rescued by whole body SIRT6 overexpression. Since the liver is the main gluconeogenic organ, we performed here liver metabolomics in young (6 months) versus old (20-24 months) WT and SIRT6-transgenic mice. We used liver tissues of 6h morning-fasted mice, either in naïve state or 15 minutes after intraperitoneal injection of [U-13C]Lactate.
Institute:	Bar Ilan University
Department:	The Mina & Everard Goodman Faculty of Life Sciences
Laboratory:	Laboratory of Prof. Haim Cohen
Last Name:	Cohen
First Name:	Chaim
Address:	Ramat Gan, Ramat Gan, Israel, 5290002, Israel
Email:	Haim.Cohen@biu.ac.il
Phone:	+97235318383
Funding Source:	Israel Science Foundation (621/13 and 777/16), I-Core foundation (1775/12) and (41/11) and Laura and Isaac Perlmutter Foundation
Contributors:	Asael Roichman, Ifat Abramovich, Eyal Gottlieb, Haim Yosef Cohen

Subject:

Subject ID:	SU001581
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6JOlaHsd
Age Or Age Range:	6 months (young) and 20-24 months (old)
Gender:	Male
Animal Animal Supplier:	Envigo
Animal Housing:	Animal rooms were maintained at 21°C-24°C and 35%-75% relative humidity
Animal Light Cycle:	12/12-h (7am to 7pm) dark-light cycle
Animal Feed:	Altromin 1324 (chow)
Animal Water:	Ad lib
Species Group:	Mammals

Factors:

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

mb_sample_id	local_sample_id	Age	Genotype	Treatment
SA126674	93_Old_Liver_25a	Old	SIRT6-tg	N/A
SA126675	94_Old_Liver_25b	Old	SIRT6-tg	N/A
SA126676	89_Old_Liver_23c	Old	SIRT6-tg	N/A
SA126677	87_Old_Liver_23a	Old	SIRT6-tg	N/A
SA126678	83_Old_Liver_21c	Old	SIRT6-tg	N/A
SA126679	95_Old_Liver_25c	Old	SIRT6-tg	N/A
SA126680	88_Old_Liver_23b	Old	SIRT6-tg	N/A
SA126681	102_Old_Liver_28a	Old	SIRT6-tg	N/A
SA126682	109_Old_Liver_30b	Old	SIRT6-tg	N/A
SA126683	110_Old_Liver_30c	Old	SIRT6-tg	N/A
SA126684	108_Old_Liver_30a	Old	SIRT6-tg	N/A
SA126685	104_Old_Liver_28c	Old	SIRT6-tg	N/A
SA126686	103_Old_Liver_28b	Old	SIRT6-tg	N/A
SA126687	82_Old_Liver_21b	Old	SIRT6-tg	N/A
SA126688	81_Old_Liver_21a	Old	SIRT6-tg	N/A
SA126689	121_Old_Liver_34b	Old	SIRT6-tg	[U-13C]Lactate
SA126690	122_Old_Liver_34c	Old	SIRT6-tg	[U-13C]Lactate
SA126691	131_Old_Liver_37c	Old	SIRT6-tg	[U-13C]Lactate
SA126692	130_Old_Liver_37b	Old	SIRT6-tg	[U-13C]Lactate
SA126693	112_Old_Liver_31b	Old	SIRT6-tg	[U-13C]Lactate
SA126694	129_Old_Liver_37a	Old	SIRT6-tg	[U-13C]Lactate
SA126695	125_Old_Liver_35c	Old	SIRT6-tg	[U-13C]Lactate
SA126696	135_Old_Liver_39a	Old	SIRT6-tg	[U-13C]Lactate
SA126697	111_Old_Liver_31a	Old	SIRT6-tg	[U-13C]Lactate
SA126698	113_Old_Liver_31c	Old	SIRT6-tg	[U-13C]Lactate
SA126699	123_Old_Liver_35a	Old	SIRT6-tg	[U-13C]Lactate
SA126700	124_Old_Liver_35b	Old	SIRT6-tg	[U-13C]Lactate
SA126701	136_Old_Liver_39b	Old	SIRT6-tg	[U-13C]Lactate
SA126702	137_Old_Liver_39c	Old	SIRT6-tg	[U-13C]Lactate
SA126703	120_Old_Liver_34a	Old	SIRT6-tg	[U-13C]Lactate
SA126704	105_Old_Liver_29a	Old	Wild-type	N/A
SA126705	107_Old_Liver_29c	Old	Wild-type	N/A
SA126706	106_Old_Liver_29b	Old	Wild-type	N/A
SA126707	90_Old_Liver_24a	Old	Wild-type	N/A
SA126708	101_Old_Liver_27c	Old	Wild-type	N/A
SA126709	98_Old_Liver_26c	Old	Wild-type	N/A
SA126710	86_Old_Liver_22c	Old	Wild-type	N/A
SA126711	92_Old_Liver_24c	Old	Wild-type	N/A
SA126712	91_Old_Liver_24b	Old	Wild-type	N/A
SA126713	85_Old_Liver_22b	Old	Wild-type	N/A
SA126714	84_Old_Liver_22a	Old	Wild-type	N/A
SA126715	99_Old_Liver_27a	Old	Wild-type	N/A
SA126716	97_Old_Liver_26b	Old	Wild-type	N/A
SA126717	96_Old_Liver_26a	Old	Wild-type	N/A
SA126718	100_Old_Liver_27b	Old	Wild-type	N/A
SA126719	117_Old_Liver_33a	Old	Wild-type	[U-13C]Lactate
SA126720	132_Old_Liver_38a	Old	Wild-type	[U-13C]Lactate
SA126721	128_Old_Liver_36c	Old	Wild-type	[U-13C]Lactate
SA126722	114_Old_Liver_32a	Old	Wild-type	[U-13C]Lactate
SA126723	133_Old_Liver_38b	Old	Wild-type	[U-13C]Lactate
SA126724	134_Old_Liver_38c	Old	Wild-type	[U-13C]Lactate
SA126725	140_Old_Liver_40c	Old	Wild-type	[U-13C]Lactate
SA126726	139_Old_Liver_40b	Old	Wild-type	[U-13C]Lactate
SA126727	138_Old_Liver_40a	Old	Wild-type	[U-13C]Lactate
SA126728	126_Old_Liver_36a	Old	Wild-type	[U-13C]Lactate
SA126729	127_Old_Liver_36b	Old	Wild-type	[U-13C]Lactate
SA126730	119_Old_Liver_33c	Old	Wild-type	[U-13C]Lactate
SA126731	116_Old_Liver_32c	Old	Wild-type	[U-13C]Lactate
SA126732	115_Old_Liver_32b	Old	Wild-type	[U-13C]Lactate
SA126733	118_Old_Liver_33b	Old	Wild-type	[U-13C]Lactate
SA126734	03_Young_Liver_1c	Young	SIRT6-tg	N/A
SA126735	02_Young_Liver_1b	Young	SIRT6-tg	N/A
SA126736	14_Young_Liver_4b	Young	SIRT6-tg	N/A
SA126737	06_Young_Liver_2b	Young	SIRT6-tg	N/A
SA126738	13_Young_Liver_4a	Young	SIRT6-tg	N/A
SA126739	07_Young_Liver_2c	Young	SIRT6-tg	N/A
SA126740	05_Young_Liver_2a	Young	SIRT6-tg	N/A
SA126741	15_Young_Liver_4c	Young	SIRT6-tg	N/A
SA126742	38_Young_Liver_10b	Young	SIRT6-tg	N/A
SA126743	01_Young_Liver_1a	Young	SIRT6-tg	N/A
SA126744	37_Young_Liver_10a	Young	SIRT6-tg	N/A
SA126745	39_Young_Liver_10c	Young	SIRT6-tg	N/A
SA126746	34_Young_Liver_9b	Young	SIRT6-tg	N/A
SA126747	33_Young_Liver_9a	Young	SIRT6-tg	N/A
SA126748	62_Young_Liver_16b	Young	SIRT6-tg	[U-13C]Lactate
SA126749	63_Young_Liver_16c	Young	SIRT6-tg	[U-13C]Lactate
SA126750	66_Young_Liver_17b	Young	SIRT6-tg	[U-13C]Lactate
SA126751	61_Young_Liver_16a	Young	SIRT6-tg	[U-13C]Lactate
SA126752	65_Young_Liver_17a	Young	SIRT6-tg	[U-13C]Lactate
SA126753	54_Young_Liver_14b	Young	SIRT6-tg	[U-13C]Lactate
SA126754	53_Young_Liver_14a	Young	SIRT6-tg	[U-13C]Lactate
SA126755	67_Young_Liver_17c	Young	SIRT6-tg	[U-13C]Lactate
SA126756	55_Young_Liver_14c	Young	SIRT6-tg	[U-13C]Lactate
SA126757	57_Young_Liver_15a	Young	SIRT6-tg	[U-13C]Lactate
SA126758	59_Young_Liver_15c	Young	SIRT6-tg	[U-13C]Lactate
SA126759	58_Young_Liver_15b	Young	SIRT6-tg	[U-13C]Lactate
SA126760	69_Young_Liver_18a	Young	SIRT6-tg	[U-13C]Lactate
SA126761	71_Young_Liver_18c	Young	SIRT6-tg	[U-13C]Lactate
SA126762	70_Young_Liver_18b	Young	SIRT6-tg	[U-13C]Lactate
SA126763	17_Young_Liver_5a	Young	Wild-type	N/A
SA126764	31_Young_Liver_8c	Young	Wild-type	N/A
SA126765	18_Young_Liver_5b	Young	Wild-type	N/A
SA126766	11_Young_Liver_3c	Young	Wild-type	N/A
SA126767	21_Young_Liver_6a	Young	Wild-type	N/A
SA126768	30_Young_Liver_8b	Young	Wild-type	N/A
SA126769	19_Young_Liver_5c	Young	Wild-type	N/A
SA126770	27_Young_Liver_7c	Young	Wild-type	N/A
SA126771	09_Young_Liver_3a	Young	Wild-type	N/A
SA126772	25_Young_Liver_7a	Young	Wild-type	N/A
SA126773	23_Young_Liver_6c	Young	Wild-type	N/A

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Collection:

Collection ID:	CO001576
Collection Summary:	Mice were euthanized by cervical dislocation, left liver lobe was collected as fast as possible (<30 sec) and immediately flash-frozen in liquid nitrogen. Tissues were stored at -80C until metabolite extraction.
Sample Type:	Liver
Storage Conditions:	-80℃

Treatment:

Treatment ID:	TR001596
Treatment Summary:	All mice were fasted for 6h (8AM-2PM). Before sacrifice, tail blood was collected from all mice for less than 2 minutes. The untreated group were sacrificed at this point. For 13C-lactate treatment, 6h fasted mice were intraperitoneally injected with 1mg/kg [U-13C]Lactate (CIL) and sacrificed 15 minutes after injection.
Treatment:	[U-13C]Lactate injection
Treatment Compound:	[U-13C]Lactate
Treatment Route:	Intraperitoneal injection
Treatment Dose:	1mg/kg
Treatment Dosevolume:	300ul for 30gr mouse
Animal Anesthesia:	N/A
Animal Fasting:	6h morning fast
Animal Endp Euthanasia:	Cervical dislocation

Sample Preparation:

Sampleprep ID:	SP001589
Sampleprep Summary:	To maximize accuracy and to take into account tissue inhomogeneity, metabolites were extracted from 3 separate liver pieces for each mouse, and the average metabolite value of the 3 technical replicates was calculated at analysis. In detail, frozen liver pieces weighting ~30mg were transferred into soft tissue homogenizing CK 14 tubes containing 1.4 mm ceramic beads (Bertin corp.) prefilled with 1 ml of cold (-20°C) metabolites extraction solvent (Methanol: Acetonitrile:H2O, 50:30:20) and kept on ice. Samples were homogenized using precellys 24 tissue homogenizer (3X20sec at 6000 rpm, with a 30sec gap between each of the three cycles, Bertin Technologies) cooled to 2°C. Homogenized extracts were centrifuged in the Precellys tubes at 18,000g for 15 minutes at 4°C, supernatants were collected in microcentrifuge tubes and centrifuged again at 18,000g for 10 minutes at 4°C. The supernatants were transferred to glass HPLC vials and kept at -75 °C prior to LC-MS analysis.

Sampleprep ID:

SP001589

Sampleprep Summary:

To maximize accuracy and to take into account tissue inhomogeneity, metabolites were extracted from 3 separate liver pieces for each mouse, and the average metabolite value of the 3 technical replicates was calculated at analysis. In detail, frozen liver pieces weighting ~30mg were transferred into soft tissue homogenizing CK 14 tubes containing 1.4 mm ceramic beads (Bertin corp.) prefilled with 1 ml of cold (-20°C) metabolites extraction solvent (Methanol: Acetonitrile:H2O, 50:30:20) and kept on ice. Samples were homogenized using precellys 24 tissue homogenizer (3X20sec at 6000 rpm, with a 30sec gap between each of the three cycles, Bertin Technologies) cooled to 2°C. Homogenized extracts were centrifuged in the Precellys tubes at 18,000g for 15 minutes at 4°C, supernatants were collected in microcentrifuge tubes and centrifuged again at 18,000g for 10 minutes at 4°C. The supernatants were transferred to glass HPLC vials and kept at -75 °C prior to LC-MS analysis.

Chromatography:

Chromatography ID:	CH001827
Instrument Name:	Thermo Dionex Ultimate 3000
Column Name:	SeQuant ZIC-HILIC (150 x 2.1mm,5um)
Column Temperature:	45
Flow Rate:	0.2 ml/min
Solvent A:	100% water; 20 mM ammonium carbonate adjusted to pH 9.2; 0.1% of 25% ammonium hydroxide
Solvent B:	100% acetonitrile
Chromatography Type:	HILIC

Analysis:

Analysis ID:	AN002498
Laboratory Name:	Prof. Eyal Gottlieb
Analysis Type:	MS
Operator Name:	Ifat Abramovich
Chromatography ID:	CH001827
Num Factors:	8
Num Metabolites:	302
Units:	Peak Area