Summary of Study ST002167

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 PR001376. The data can be accessed directly via it's Project DOI: 10.21228/M8VH8V 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 IDST002167
Study TitleRemote solid cancers rewire hepatic nitrogen metabolism via host nicotinamide-N-methyltransferase (AML cells)
Study SummaryCancers disrupt host homeostasis in various manners but the identity of host factors underlying such disruption remains largely unknown. Here we show that nicotinamide-N-methyltransferase (NNMT) is a novel host factor that mediates metabolic dysfunction in the livers of cancer-bearing mice. Multiple solid cancers distantly increase expression of Nnmt and its product 1-methylnicotinamide (MNAM) in the liver. Multi-omics analyses reveal suppression of the urea cycle accompanied by accumulation of amino acids, and enhancement of uracil biogenesis in the livers of cancer-bearing mice. Importantly, genetic deletion of Nnmt leads to alleviation of these metabolic abnormalities, and buffers cancer-dependent weight loss and reduction of the voluntary wheel-running activity. Our data also demonstrate that MNAM is capable of affecting urea cycle metabolites in the liver. These results suggest that cancers up-regulate the hepatic NNMT pathway to rewire liver metabolism towards uracil biogenesis rather than nitrogen disposal via the urea cycle, thereby disrupting host homeostasis. Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HR/MS/MS. Cationic polar metabolites (i.e., amino acids, bases, nucleosides, NAM, SAM, MNAM, SAH, me2PY, me4PY, etc) were analyzed via PFPP-LC/HR/MS/MS.
Institute
Tohoku University
Last NameKawaoka
First NameShinpei
Address4-1 Seiryo-cho, Sendai, Miyagi, 9808575, Japan
Emailkawaokashinpei@gmail.com
Phone0227178568
Submit Date2022-05-13
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailLC-MS
Release Date2022-06-01
Release Version1
Shinpei Kawaoka Shinpei Kawaoka
https://dx.doi.org/10.21228/M8VH8V
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR001376
Project DOI:doi: 10.21228/M8VH8V
Project Title:Remote solid cancers rewire hepatic nitrogen metabolism via host nicotinamide-N-methyltransferase
Project Summary:Cancers disrupt host homeostasis in various manners but the identity of host factors underlying such disruption remains largely unknown. Here we show that nicotinamide-N-methyltransferase (NNMT) is a novel host factor that mediates metabolic dysfunction in the livers of cancer-bearing mice. Multiple solid cancers distantly increase expression of Nnmt and its product 1-methylnicotinamide (MNAM) in the liver. Multi-omics analyses reveal suppression of the urea cycle accompanied by accumulation of amino acids, and enhancement of uracil biogenesis in the livers of cancer-bearing mice. Importantly, genetic deletion of Nnmt leads to alleviation of these metabolic abnormalities, and buffers cancer-dependent weight loss and reduction of the voluntary wheel-running activity. Our data also demonstrate that MNAM is capable of affecting urea cycle metabolites in the liver. These results suggest that cancers up-regulate the hepatic NNMT pathway to rewire liver metabolism towards uracil biogenesis rather than nitrogen disposal via the urea cycle, thereby disrupting host homeostasis.
Institute:Tohoku University
Last Name:Kawaoka
First Name:Shinpei
Address:4-1 Seiryo-cho, Sendai, Miyagi, 9808575, Japan
Email:kawaokashinpei@gmail.com
Phone:0227178568

Subject:

Subject ID:SU002253
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 Culture conditions Treatment
SA2076214T1_14T1-conditioned media No treatment
SA2076224T1_34T1-conditioned media No treatment
SA2076234T1_44T1-conditioned media No treatment
SA2076244T1_24T1-conditioned media No treatment
SA207625cont_2Control No treatment
SA207626cont_3Control No treatment
SA207627control_1Control No treatment
SA207628cont_1Control No treatment
SA207629control_2Control No treatment
SA207630control_3Control No treatment
SA207631control_4Control No treatment
SA207635TNF200_3Control TNFalpha (200 ng/mL) treatment
SA207636TNF200_2Control TNFalpha (200 ng/mL) treatment
SA207637TNF200_1Control TNFalpha (200 ng/mL) treatment
SA207632TNF20_3Control TNFalpha (20 ng/mL) treatment
SA207633TNF20_2Control TNFalpha (20 ng/mL) treatment
SA207634TNF20_1Control TNFalpha (20 ng/mL) treatment
Showing results 1 to 17 of 17

Collection:

Collection ID:CO002246
Collection Summary:4T1 cells were cultured in 10 cm dishes for 48 hours and the culture supernatant was collected. The supernatant was stored as the 4T1-conditioned media at 4°C until use. AML cells per a well were cultured in a 6 well plate for 24 hours, and then the media was switched to the 4T1-conditioned media. After 24 hours, the treated AML12 cells were collected.
Sample Type:AML cells

Treatment:

Treatment ID:TR002265
Treatment Summary:AML cells per well were cultured in a 24 well plate for 24 hours. The media was then switched to the bovine-serum free media, and TNF alpha was added at the concentration of 20 ng/ml or 200 ng/ml (Roche). After 24 hours, the treated AML12 cells were collected.

Sample Preparation:

Sampleprep ID:SP002259
Sampleprep Summary:Metabolites were extracted from AML12 cells (less than 6 × 10E5 cells/well (6 well plate)) using the Bligh and Dyer’s method with some modifications. Briefly, each sample was mixed with 1 mL of cold methanol containing 10-camphorsulfonic acid (1.5 nmol) and piperazine-1,4-bis (2-ethanesulfonic acid) (PIPES, 1.5 nmol) as internal standards for mass spectrometry-based metabolomic analysis. The samples were vigorously mixed by vortexing for 1 min followed by 5 min of sonication. The extracts were then centrifuged at 16,000 × g for 5 min at 4 °C, and the resultant supernatant (400 uL) was collected. After mixing 400 uL of supernatant with 400 uL of chloroform and 320 uL of water, the aqueous and organic layers were separated by vortexing and subsequent centrifugation at 16,000 × g and 4 °C for 5 min. The aqueous (upper) layer (500 uL) was transferred into a clean tube. After the aqueous layer extracts were evaporated under vacuum, the dried extracts were stored at −80 °C until the analysis of hydrophilic metabolites. Prior to analysis, the dried aqueous layer was reconstituted in 50 uL of water.

Combined analysis:

Analysis ID AN003550 AN003551
Analysis type MS MS
Chromatography type Ion exchange Reversed phase
Chromatography system Thermo Dionex ICS-5000+ Shimadzu Nexera X2
Column Dionex IonPac AS11-HC (2 um i.d. × 250 mm, 4 um particle size) Discovery HS (2.1 mm i.d. × 150 mm, 3 um particle size, Merck)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode NEGATIVE POSITIVE
Units peak area peak area

Chromatography:

Chromatography ID:CH002622
Chromatography Summary:Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HRMS/MS.
Instrument Name:Thermo Dionex ICS-5000+
Column Name:Dionex IonPac AS11-HC (2 um i.d. × 250 mm, 4 um particle size)
Chromatography Type:Ion exchange
  
Chromatography ID:CH002623
Chromatography Summary:Cationic polar metabolites (i.e., amino acids, bases, nucleosides, NAM, SAM, MNAM, SAH, me2PY, me4PY, etc) were analyzed via PFPP-LC/HRMS/MS.
Instrument Name:Shimadzu Nexera X2
Column Name:Discovery HS (2.1 mm i.d. × 150 mm, 3 um particle size, Merck)
Chromatography Type:Reversed phase

MS:

MS ID:MS003307
Analysis ID:AN003550
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:-
Ion Mode:NEGATIVE
  
MS ID:MS003308
Analysis ID:AN003551
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:-
Ion Mode:POSITIVE
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