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.

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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.

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Study IDST001507
Study TitleHepatic [U-13C]Lactate tracing and metabolomics in young and old WT and SIRT6 overexpressing mice
Study SummaryOur 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
DepartmentThe Mina & Everard Goodman Faculty of Life Sciences
LaboratoryLaboratory of Prof. Haim Cohen
Last NameCohen
First NameChaim
AddressBar Ilan University, Ramat Gan, Israel, 5290002
EmailHaim.Cohen@biu.ac.il
Phone+97235318383
Submit Date2020-10-06
Num Groups8
Total Subjects40
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2021-03-11
Release Version1
Chaim Cohen Chaim Cohen
https://dx.doi.org/10.21228/M87M4S
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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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

Factors:

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

mb_sample_id local_sample_id Age Genotype Treatment
SA12667493_Old_Liver_25aOld SIRT6-tg N/A
SA12667594_Old_Liver_25bOld SIRT6-tg N/A
SA12667689_Old_Liver_23cOld SIRT6-tg N/A
SA12667787_Old_Liver_23aOld SIRT6-tg N/A
SA12667883_Old_Liver_21cOld SIRT6-tg N/A
SA12667995_Old_Liver_25cOld SIRT6-tg N/A
SA12668088_Old_Liver_23bOld SIRT6-tg N/A
SA126681102_Old_Liver_28aOld SIRT6-tg N/A
SA126682109_Old_Liver_30bOld SIRT6-tg N/A
SA126683110_Old_Liver_30cOld SIRT6-tg N/A
SA126684108_Old_Liver_30aOld SIRT6-tg N/A
SA126685104_Old_Liver_28cOld SIRT6-tg N/A
SA126686103_Old_Liver_28bOld SIRT6-tg N/A
SA12668782_Old_Liver_21bOld SIRT6-tg N/A
SA12668881_Old_Liver_21aOld SIRT6-tg N/A
SA126689121_Old_Liver_34bOld SIRT6-tg [U-13C]Lactate
SA126690122_Old_Liver_34cOld SIRT6-tg [U-13C]Lactate
SA126691131_Old_Liver_37cOld SIRT6-tg [U-13C]Lactate
SA126692130_Old_Liver_37bOld SIRT6-tg [U-13C]Lactate
SA126693112_Old_Liver_31bOld SIRT6-tg [U-13C]Lactate
SA126694129_Old_Liver_37aOld SIRT6-tg [U-13C]Lactate
SA126695125_Old_Liver_35cOld SIRT6-tg [U-13C]Lactate
SA126696135_Old_Liver_39aOld SIRT6-tg [U-13C]Lactate
SA126697111_Old_Liver_31aOld SIRT6-tg [U-13C]Lactate
SA126698113_Old_Liver_31cOld SIRT6-tg [U-13C]Lactate
SA126699123_Old_Liver_35aOld SIRT6-tg [U-13C]Lactate
SA126700124_Old_Liver_35bOld SIRT6-tg [U-13C]Lactate
SA126701136_Old_Liver_39bOld SIRT6-tg [U-13C]Lactate
SA126702137_Old_Liver_39cOld SIRT6-tg [U-13C]Lactate
SA126703120_Old_Liver_34aOld SIRT6-tg [U-13C]Lactate
SA126704105_Old_Liver_29aOld Wild-type N/A
SA126705107_Old_Liver_29cOld Wild-type N/A
SA126706106_Old_Liver_29bOld Wild-type N/A
SA12670790_Old_Liver_24aOld Wild-type N/A
SA126708101_Old_Liver_27cOld Wild-type N/A
SA12670998_Old_Liver_26cOld Wild-type N/A
SA12671086_Old_Liver_22cOld Wild-type N/A
SA12671192_Old_Liver_24cOld Wild-type N/A
SA12671291_Old_Liver_24bOld Wild-type N/A
SA12671385_Old_Liver_22bOld Wild-type N/A
SA12671484_Old_Liver_22aOld Wild-type N/A
SA12671599_Old_Liver_27aOld Wild-type N/A
SA12671697_Old_Liver_26bOld Wild-type N/A
SA12671796_Old_Liver_26aOld Wild-type N/A
SA126718100_Old_Liver_27bOld Wild-type N/A
SA126719117_Old_Liver_33aOld Wild-type [U-13C]Lactate
SA126720132_Old_Liver_38aOld Wild-type [U-13C]Lactate
SA126721128_Old_Liver_36cOld Wild-type [U-13C]Lactate
SA126722114_Old_Liver_32aOld Wild-type [U-13C]Lactate
SA126723133_Old_Liver_38bOld Wild-type [U-13C]Lactate
SA126724134_Old_Liver_38cOld Wild-type [U-13C]Lactate
SA126725140_Old_Liver_40cOld Wild-type [U-13C]Lactate
SA126726139_Old_Liver_40bOld Wild-type [U-13C]Lactate
SA126727138_Old_Liver_40aOld Wild-type [U-13C]Lactate
SA126728126_Old_Liver_36aOld Wild-type [U-13C]Lactate
SA126729127_Old_Liver_36bOld Wild-type [U-13C]Lactate
SA126730119_Old_Liver_33cOld Wild-type [U-13C]Lactate
SA126731116_Old_Liver_32cOld Wild-type [U-13C]Lactate
SA126732115_Old_Liver_32bOld Wild-type [U-13C]Lactate
SA126733118_Old_Liver_33bOld Wild-type [U-13C]Lactate
SA12673403_Young_Liver_1cYoung SIRT6-tg N/A
SA12673502_Young_Liver_1bYoung SIRT6-tg N/A
SA12673614_Young_Liver_4bYoung SIRT6-tg N/A
SA12673706_Young_Liver_2bYoung SIRT6-tg N/A
SA12673813_Young_Liver_4aYoung SIRT6-tg N/A
SA12673907_Young_Liver_2cYoung SIRT6-tg N/A
SA12674005_Young_Liver_2aYoung SIRT6-tg N/A
SA12674115_Young_Liver_4cYoung SIRT6-tg N/A
SA12674238_Young_Liver_10bYoung SIRT6-tg N/A
SA12674301_Young_Liver_1aYoung SIRT6-tg N/A
SA12674437_Young_Liver_10aYoung SIRT6-tg N/A
SA12674539_Young_Liver_10cYoung SIRT6-tg N/A
SA12674634_Young_Liver_9bYoung SIRT6-tg N/A
SA12674733_Young_Liver_9aYoung SIRT6-tg N/A
SA12674862_Young_Liver_16bYoung SIRT6-tg [U-13C]Lactate
SA12674963_Young_Liver_16cYoung SIRT6-tg [U-13C]Lactate
SA12675066_Young_Liver_17bYoung SIRT6-tg [U-13C]Lactate
SA12675161_Young_Liver_16aYoung SIRT6-tg [U-13C]Lactate
SA12675265_Young_Liver_17aYoung SIRT6-tg [U-13C]Lactate
SA12675354_Young_Liver_14bYoung SIRT6-tg [U-13C]Lactate
SA12675453_Young_Liver_14aYoung SIRT6-tg [U-13C]Lactate
SA12675567_Young_Liver_17cYoung SIRT6-tg [U-13C]Lactate
SA12675655_Young_Liver_14cYoung SIRT6-tg [U-13C]Lactate
SA12675757_Young_Liver_15aYoung SIRT6-tg [U-13C]Lactate
SA12675859_Young_Liver_15cYoung SIRT6-tg [U-13C]Lactate
SA12675958_Young_Liver_15bYoung SIRT6-tg [U-13C]Lactate
SA12676069_Young_Liver_18aYoung SIRT6-tg [U-13C]Lactate
SA12676171_Young_Liver_18cYoung SIRT6-tg [U-13C]Lactate
SA12676270_Young_Liver_18bYoung SIRT6-tg [U-13C]Lactate
SA12676317_Young_Liver_5aYoung Wild-type N/A
SA12676431_Young_Liver_8cYoung Wild-type N/A
SA12676518_Young_Liver_5bYoung Wild-type N/A
SA12676611_Young_Liver_3cYoung Wild-type N/A
SA12676721_Young_Liver_6aYoung Wild-type N/A
SA12676830_Young_Liver_8bYoung Wild-type N/A
SA12676919_Young_Liver_5cYoung Wild-type N/A
SA12677027_Young_Liver_7cYoung Wild-type N/A
SA12677109_Young_Liver_3aYoung Wild-type N/A
SA12677225_Young_Liver_7aYoung Wild-type N/A
SA12677323_Young_Liver_6cYoung 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.

Combined analysis:

Analysis ID AN002498
Analysis type MS
Chromatography type HILIC
Chromatography system Thermo Dionex Ultimate 3000
Column SeQuant ZIC-HILIC (150 x 2.1mm,5um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode UNSPECIFIED
Units Peak Area

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

MS:

MS ID:MS002318
Analysis ID:AN002498
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:polarity switching mode to enable both positive and negative ions was used
Ion Mode:UNSPECIFIED
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