Summary of Study ST002163

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.

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	ST002163
Study Title	Remote solid cancers rewire hepatic nitrogen metabolism via host nicotinamide-N-methyltransferase
Study 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. 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 Name	Kawaoka
First Name	Shinpei
Address	4-1, Seiryou-machi, Aoba-ku, Sendai
Email	kawaokashinpei@gmail.com
Phone	0227178443
Submit Date	2022-05-11
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2022-05-27
Release Version	1

Select appropriate tab below to view additional metadata details:

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:	SU002249
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Species Group:	Mammals

Factors:

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

mb_sample_id	local_sample_id	Genotype	Sample group	Sex	Treatment
SA207523	L14	NNMT KO	4T1	female	No treatment
SA207524	L15	NNMT KO	4T1	female	No treatment
SA207525	L51	NNMT KO	4T1	female	No treatment
SA207526	L13	NNMT KO	4T1	female	No treatment
SA207527	L16	NNMT KO	4T1	female	No treatment
SA207528	L11	NNMT KO	Sham	female	No treatment
SA207529	L12	NNMT KO	Sham	female	No treatment
SA207530	L52	NNMT KO	Sham	female	No treatment
SA207531	L10	NNMT KO	Sham	female	No treatment
SA207532	L9	NNMT KO	Sham	female	No treatment
SA207533	L13_MNAM	WT	4T1	female	MNAM treatment
SA207534	L14_MNAM	WT	4T1	female	MNAM treatment
SA207535	L15_MNAM	WT	4T1	female	MNAM treatment
SA207536	L16_MNAM	WT	4T1	female	MNAM treatment
SA207537	L17_MNAM	WT	4T1	female	MNAM treatment
SA207538	L12_MNAM	WT	4T1	female	No treatment
SA207539	L9_MNAM	WT	4T1	female	No treatment
SA207540	L10_MNAM	WT	4T1	female	No treatment
SA207541	L6	WT	4T1	female	No treatment
SA207542	L7	WT	4T1	female	No treatment
SA207543	L8	WT	4T1	female	No treatment
SA207544	L5	WT	4T1	female	No treatment
SA207545	L50	WT	4T1	female	No treatment
SA207546	L11_MNAM	WT	4T1	female	No treatment
SA207547	L8_MNAM	WT	Sham	female	MNAM treatment
SA207548	L6_MNAM	WT	Sham	female	MNAM treatment
SA207549	L7_MNAM	WT	Sham	female	MNAM treatment
SA207550	L5_MNAM	WT	Sham	female	MNAM treatment
SA207551	L2	WT	Sham	female	No treatment
SA207552	L2_MNAM	WT	Sham	female	No treatment
SA207553	L4	WT	Sham	female	No treatment
SA207554	L49	WT	Sham	female	No treatment
SA207555	L1	WT	Sham	female	No treatment
SA207556	L3	WT	Sham	female	No treatment
SA207557	L3_MNAM	WT	Sham	female	No treatment
SA207558	L1_MNAM	WT	Sham	female	No treatment
SA207559	L4_MNAM	WT	Sham	female	No treatment

Showing results 1 to 37 of 37

Showing results 1 to 37 of 37

Collection:

Collection ID:	CO002242
Collection Summary:	Mouse livers were crushed in liquid-nitrogen and homogenized.
Sample Type:	Liver

Treatment:

Treatment ID:	TR002261
Treatment Summary:	Daily 250 mg/kg MNAM injection for 12 days in the absence (sham) and presence (4T1cancers).

Sample Preparation:

Sampleprep ID:	SP002255
Sampleprep Summary:	Metabolites were extracted from the frozen livers (approximately 5 mg) 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.

Sampleprep ID:

SP002255

Sampleprep Summary:

Metabolites were extracted from the frozen livers (approximately 5 mg) 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	AN003544	AN003545
Analysis type	MS	MS
Chromatography type	Ion exchange	Reversed phase
Chromatography system	Dionex ICS-5000+ HPIC system	Shimadzu Nexera X2
Column	Dionex IonPac AS11-HC (250 x 2mm,4um)	Discovery HS (150 x 2.1mm,3um)
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:	CH002618
Chromatography Summary:	Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HRMS/MS.
Instrument Name:	Dionex ICS-5000+ HPIC system
Column Name:	Dionex IonPac AS11-HC (250 x 2mm,4um)
Chromatography Type:	Ion exchange

Chromatography ID:	CH002619
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 (150 x 2.1mm,3um)
Chromatography Type:	Reversed phase

MS:

MS ID:	MS003302
Analysis ID:	AN003544
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	-
Ion Mode:	NEGATIVE

MS ID:	MS003303
Analysis ID:	AN003545
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	-
Ion Mode:	POSITIVE