{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002163","ANALYSIS_ID":"AN003544","VERSION":"1","CREATED_ON":"May 13, 2022, 7:22 am"},

"PROJECT":{"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"},

"STUDY":{"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"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"190428",
"Sample ID":"L1",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L1"}
},
{
"Subject ID":"190428",
"Sample ID":"L2",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L2"}
},
{
"Subject ID":"190428",
"Sample ID":"L3",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L3"}
},
{
"Subject ID":"190428",
"Sample ID":"L4",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L4"}
},
{
"Subject ID":"190428",
"Sample ID":"L49",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L49"}
},
{
"Subject ID":"190428",
"Sample ID":"L5",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L5"}
},
{
"Subject ID":"190428",
"Sample ID":"L6",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L6"}
},
{
"Subject ID":"190428",
"Sample ID":"L7",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L7"}
},
{
"Subject ID":"190428",
"Sample ID":"L8",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L8"}
},
{
"Subject ID":"190428",
"Sample ID":"L50",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L50"}
},
{
"Subject ID":"190428",
"Sample ID":"L9",
"Factors":{"Genotype":"NNMT KO","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L9"}
},
{
"Subject ID":"190428",
"Sample ID":"L10",
"Factors":{"Genotype":"NNMT KO","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L10"}
},
{
"Subject ID":"190428",
"Sample ID":"L11",
"Factors":{"Genotype":"NNMT KO","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L11"}
},
{
"Subject ID":"190428",
"Sample ID":"L12",
"Factors":{"Genotype":"NNMT KO","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L12"}
},
{
"Subject ID":"190428",
"Sample ID":"L52",
"Factors":{"Genotype":"NNMT KO","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L52"}
},
{
"Subject ID":"190428",
"Sample ID":"L13",
"Factors":{"Genotype":"NNMT KO","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L13"}
},
{
"Subject ID":"190428",
"Sample ID":"L14",
"Factors":{"Genotype":"NNMT KO","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L14"}
},
{
"Subject ID":"190428",
"Sample ID":"L15",
"Factors":{"Genotype":"NNMT KO","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L15"}
},
{
"Subject ID":"190428",
"Sample ID":"L16",
"Factors":{"Genotype":"NNMT KO","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L16"}
},
{
"Subject ID":"190428",
"Sample ID":"L51",
"Factors":{"Genotype":"NNMT KO","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L51"}
},
{
"Subject ID":"201215",
"Sample ID":"L1_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L1_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L2_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L2_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L3_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L3_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L4_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L4_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L5_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L5_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L6_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L6_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L7_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L7_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L8_MNAM",
"Factors":{"Genotype":"WT","Sample group":"Sham","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L8_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L9_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L9_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L10_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L10_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L11_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L11_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L12_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"No treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L12_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L13_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L13_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L14_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L14_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L15_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L15_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L16_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L16_MNAM"}
},
{
"Subject ID":"201215",
"Sample ID":"L17_MNAM",
"Factors":{"Genotype":"WT","Sample group":"4T1","Sex":"female","Treatment":"MNAM treatment"},
"Additional sample data":{"RAW_FILE_NAME":"L17_MNAM"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mouse livers were crushed in liquid-nitrogen and homogenized.","SAMPLE_TYPE":"Liver"},

"TREATMENT":{"TREATMENT_SUMMARY":"Daily 250 mg/kg MNAM injection for 12 days in the absence (sham) and presence (4T1cancers)."},

"SAMPLEPREP":{"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."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Anionic polar metabolites (i.e., organic acids, sugar phosphates, nucleotides, etc.) were analyzed via IC/HRMS/MS.","CHROMATOGRAPHY_TYPE":"Other","INSTRUMENT_NAME":"Dionex ICS-5000+ HPIC system","COLUMN_NAME":"Dionex IonPac AS11-HC (2 um i.d. × 250 mm, 4 um particle size)"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","LABORATORY_NAME":"Bamba lab.","OPERATOR_NAME":"Masatomo Takahashi"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"-"},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"10-Camphorsulfonic acid (IS)","L1":"5.E+08","L2":"6.E+08","L3":"3.E+08","L4":"4.E+08","L49":"3.E+08","L5":"2.E+08","L6":"3.E+08","L7":"5.E+08","L8":"2.E+08","L50":"2.E+08","L9":"2.E+08","L10":"2.E+08","L11":"3.E+08","L12":"3.E+08","L52":"2.E+08","L13":"4.E+08","L14":"5.E+08","L15":"4.E+08","L16":"3.E+08","L51":"2.E+08","L1_MNAM":"5.E+08","L2_MNAM":"4.E+08","L3_MNAM":"5.E+08","L4_MNAM":"4.E+08","L5_MNAM":"4.E+08","L6_MNAM":"4.E+08","L7_MNAM":"4.E+08","L8_MNAM":"5.E+08","L9_MNAM":"4.E+08","L10_MNAM":"4.E+08","L11_MNAM":"4.E+08","L12_MNAM":"4.E+08","L13_MNAM":"4.E+08","L14_MNAM":"4.E+08","L15_MNAM":"4.E+08","L16_MNAM":"4.E+08","L17_MNAM":"4.E+08"},{"Metabolite":"PIPES 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}

}