{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002376","ANALYSIS_ID":"AN003871","VERSION":"1","CREATED_ON":"December 1, 2022, 9:57 am"},

"PROJECT":{"PROJECT_TITLE":"Hepatic Phosphatidylcholine Catabolism Driven by PNPLA7 and PNPLA8 Supplies Endogenous Choline to Replenish the Methionine Cycle with Methyl Groups","PROJECT_SUMMARY":"Choline supplies methyl groups for regeneration of methionine and the methyl donor S-adenosylmethionine in the liver. Here we demonstrate that the catabolism of membrane phosphatidylcholine (PC) into water-soluble glycerophosphocholine (GPC) by the phospholipase/lysophospholipase PNPLA8-PNPLA7 axis enables endogenous choline stored in hepatic PC to be utilized in methyl metabolism. PNPLA7-deficient mice show marked decreases in hepatic GPC, choline, and several metabolites related to the methionine cycle, accompanied by various signs of methionine insufficiency including growth retardation, hypoglycemia, hypolipidemia, increased energy consumption, reduced adiposity, increased FGF21, and an altered histone/DNA methylation landscape. Moreover, PNPLA8-deficient mice recapitulate most of these phenotypes. In contrast to wild-type mice fed a methionine/choline-deficient diet, both knockout strains display a decreased hepatic triglyceride likely via reductions of lipogenesis and GPC-derived glycerol flux. Collectively, our findings highlight the biological importance of phospholipid catabolism driven by PNPLA8/PNPLA7 in methyl group flux and triglyceride synthesis in the liver.","INSTITUTE":"Tokyo Metropolitan Institute of Medical Science","LAST_NAME":"Hirabayashi","FIRST_NAME":"Tetsuya","ADDRESS":"2-6-1 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan","EMAIL":"hirabayashi-tt@igakuken.or.jp","PHONE":"+81-3-5316-3100"},

"STUDY":{"STUDY_TITLE":"Hepatic Phosphatidylcholine Catabolism Driven by PNPLA7 and PNPLA8 Supplies Endogenous Choline to Replenish the Methionine Cycle with Methyl Groups(Pnpla8-knockout)","STUDY_SUMMARY":"Choline supplies methyl groups for regeneration of methionine and the methyl donor S-adenosylmethionine in the liver. Here we demonstrate that the catabolism of membrane phosphatidylcholine (PC) into water-soluble glycerophosphocholine (GPC) by the phospholipase/lysophospholipase PNPLA8-PNPLA7 axis enables endogenous choline stored in hepatic PC to be utilized in methyl metabolism. PNPLA7-deficient mice show marked decreases in hepatic GPC, choline, and several metabolites related to the methionine cycle, accompanied by various signs of methionine insufficiency including growth retardation, hypoglycemia, hypolipidemia, increased energy consumption, reduced adiposity, increased FGF21, and an altered histone/DNA methylation landscape. Moreover, PNPLA8-deficient mice recapitulate most of these phenotypes. In contrast to wild-type mice fed a methionine/choline-deficient diet, both knockout strains display a decreased hepatic triglyceride likely via reductions of lipogenesis and GPC-derived glycerol flux. Collectively, our findings highlight the biological importance of phospholipid catabolism driven by PNPLA8/PNPLA7 in methyl group flux and triglyceride synthesis in the liver.","INSTITUTE":"Tokyo Metropolitan Institute of Medical Science","LAST_NAME":"Hirabayashi","FIRST_NAME":"Tetsuya","ADDRESS":"2-6-1 Kamikitazawa, Setagaya-ku, Tokyo, 156-8506, Japan","EMAIL":"hirabayashi-tt@igakuken.or.jp","PHONE":"+81-3-5316-3100"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENOTYPE_STRAIN":"C57BL/6N","AGE_OR_AGE_RANGE":"6-weeks old"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"WT1",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"WT2",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"WT3",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"WT4",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"WT5",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"WT6",
"Factors":{"Genotype":"Wild-type"}
},
{
"Subject ID":"-",
"Sample ID":"KO1",
"Factors":{"Genotype":"Pnpla8-knockout"}
},
{
"Subject ID":"-",
"Sample ID":"KO2",
"Factors":{"Genotype":"Pnpla8-knockout"}
},
{
"Subject ID":"-",
"Sample ID":"KO3",
"Factors":{"Genotype":"Pnpla8-knockout"}
},
{
"Subject ID":"-",
"Sample ID":"KO4",
"Factors":{"Genotype":"Pnpla8-knockout"}
},
{
"Subject ID":"-",
"Sample ID":"KO5",
"Factors":{"Genotype":"Pnpla8-knockout"}
},
{
"Subject ID":"-",
"Sample ID":"KO6",
"Factors":{"Genotype":"Pnpla8-knockout"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Mice were deeply anesthetized and perfused intracardially with saline containing 1 mM MOPS (pH 7.4). Liver was rapidly removed, frozen on liquid nitrogen and stored at –80°C until further processing.","SAMPLE_TYPE":"Liver","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"No treatment in this study."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Tissues with internal standards were homogenized with ice-cold acetonitrile:water (50:50) and centrifuged at 2,300 × g for 5 min at 4 °C. The supernatant was filtered through a 5-kDa cut-off filter (Ultrafree-MC-PLHCC, Human Metabolome Technologies) at 9,100 × g for 2 h at 4 °C, dried in a centrifugal evaporator at 1,500 rpm at 1,000 Pa, and resuspended in 50 μl of ultrapure water before metabolome analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"CE","INSTRUMENT_NAME":"Agilent 7100 CE","COLUMN_NAME":"Fused silica capillary i.d. 50 μm × 80 cm"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Agilent CE-TOFMS","INSTRUMENT_TYPE":"Other","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The spectrometer was scanned from m/z 50 to 1,000. Peaks were extracted using automatic integration software MasterHands (ver.2.16.0.15 Keio University). Signal peaks corresponding to isotopomers, adduct ions, and other product ions of known metabolites were excluded, and remaining peaks were annotated with putative metabolites from the HMT metabolite database based on their migration time and m/z values. Peak areas were normalized to internal standards and sample amounts."},

"MS_METABOLITE_DATA":{
"Units":"Peak area",

"Data":[{"Metabolite":"1-Aminocyclopentanecarboxylic acid","WT1":"N.D.","WT2":"N.D.","WT3":"N.D.","WT4":"N.D.","WT5":"N.D.","WT6":"N.D.","KO1":"9.4E-05","KO2":"N.D.","KO3":"N.D.","KO4":"N.D.","KO5":"N.D.","KO6":"N.D."},{"Metabolite":"1-Aminocyclopropane-1-carboxylic acid","WT1":"1.3E-03","WT2":"9.5E-04","WT3":"7.8E-04","WT4":"1.1E-03","WT5":"9.3E-04","WT6":"7.9E-04","KO1":"7.8E-04","KO2":"6.9E-04","KO3":"5.2E-04","KO4":"6.2E-04","KO5":"7.7E-04","KO6":"7.8E-04"},{"Metabolite":"1-Methyl-4-imidazoleacetic acid","WT1":"7.1E-04","WT2":"6.6E-04","WT3":"5.8E-04","WT4":"1.1E-03","WT5":"3.0E-04","WT6":"2.6E-04","KO1":"4.1E-04","KO2":"3.4E-04","KO3":"3.1E-04","KO4":"2.3E-04","KO5":"3.1E-04","KO6":"5.4E-04"},{"Metabolite":"1-Methyladenosine","WT1":"6.5E-05","WT2":"1.4E-04","WT3":"1.2E-04","WT4":"1.1E-04","WT5":"8.6E-05","WT6":"6.9E-05","KO1":"1.1E-04","KO2":"1.5E-04","KO3":"1.1E-04","KO4":"7.0E-05","KO5":"8.2E-05","KO6":"1.2E-04"},{"Metabolite":"1-Methylhistamine","WT1":"7.2E-05","WT2":"1.5E-04","WT3":"9.4E-05","WT4":"6.9E-05","WT5":"N.D.","WT6":"4.5E-05","KO1":"7.0E-05","KO2":"8.1E-05","KO3":"9.0E-05","KO4":"N.D.","KO5":"N.D.","KO6":"3.9E-05"},{"Metabolite":"1-Methylnicotinamide","WT1":"5.2E-04","WT2":"4.6E-04","WT3":"5.0E-04","WT4":"3.1E-04","WT5":"1.1E-03","WT6":"9.7E-04","KO1":"1.5E-03","KO2":"1.7E-03","KO3":"1.5E-03","KO4":"1.3E-03","KO5":"1.2E-03","KO6":"1.0E-03"},{"Metabolite":"1-Pyrroline 5-carboxylic acid","WT1":"3.9E-04","WT2":"6.4E-04","WT3":"3.5E-04","WT4":"4.1E-04","WT5":"5.2E-04","WT6":"5.8E-04","KO1":"9.8E-04","KO2":"1.1E-03","KO3":"5.8E-04","KO4":"6.3E-04","KO5":"7.5E-04","KO6":"6.8E-04"},{"Metabolite":"1H-Imidazole-4-propionic acid","WT1":"N.D.","WT2":"1.6E-04","WT3":"N.D.","WT4":"6.8E-04","WT5":"2.0E-04","WT6":"4.6E-04","KO1":"6.4E-05","KO2":"4.5E-04","KO3":"1.4E-04","KO4":"3.2E-04","KO5":"3.0E-04","KO6":"1.9E-04"},{"Metabolite":"2-Amino-2-(hydroxymethyl)-1,3-propanediol","WT1":"N.D.","WT2":"N.D.","WT3":"1.1E-04","WT4":"N.D.","WT5":"2.0E-01","WT6":"3.0E-01","KO1":"9.8E-05","KO2":"N.D.","KO3":"1.0E-04","KO4":"3.2E-01","KO5":"2.1E-01","KO6":"N.D."},{"Metabolite":"2-Aminoadipic acid","WT1":"3.4E-03","WT2":"1.9E-03","WT3":"8.1E-04","WT4":"2.5E-03","WT5":"2.1E-03","WT6":"8.5E-03","KO1":"2.9E-03","KO2":"5.7E-03","KO3":"3.5E-03","KO4":"4.2E-03","KO5":"1.0E-02","KO6":"3.8E-03"},{"Metabolite":"2-Aminoisobutyric acid","WT1":"1.6E-02","WT2":"2.0E-02","WT3":"1.4E-02","WT4":"1.8E-02","WT5":"5.2E-03","WT6":"4.6E-03","KO1":"1.6E-02","KO2":"1.6E-02","KO3":"7.4E-03","KO4":"7.1E-03","KO5":"8.4E-03","KO6":"6.5E-03"},{"Metabolite":"2-Methylserine","WT1":"3.6E-04","WT2":"3.7E-04","WT3":"1.5E-04","WT4":"1.0E-03","WT5":"2.6E-04","WT6":"2.2E-04","KO1":"2.3E-04","KO2":"3.1E-04","KO3":"2.4E-04","KO4":"2.9E-04","KO5":"2.5E-04","KO6":"6.3E-04"},{"Metabolite":"2,3-Diaminopropionic acid","WT1":"N.D.","WT2":"1.3E-04","WT3":"5.0E-05","WT4":"9.5E-05","WT5":"7.6E-05","WT6":"2.1E-04","KO1":"N.D.","KO2":"N.D.","KO3":"4.4E-05","KO4":"N.D.","KO5":"9.5E-05","KO6":"N.D."},{"Metabolite":"2,4-Diaminobutyric acid","WT1":"2.8E-04","WT2":"4.3E-04","WT3":"2.0E-04","WT4":"6.1E-04","WT5":"1.8E-04","WT6":"2.7E-04","KO1":"1.5E-04","KO2":"1.2E-04","KO3":"2.5E-04","KO4":"2.3E-04","KO5":"2.7E-04","KO6":"2.5E-04"},{"Metabolite":"2,6-Diaminopimelic acid","WT1":"N.D.","WT2":"N.D.","WT3":"N.D.","WT4":"N.D.","WT5":"5.1E-05","WT6":"4.4E-05","KO1":"7.0E-05","KO2":"5.9E-05","KO3":"4.8E-05","KO4":"4.8E-05","KO5":"8.4E-05","KO6":"N.D."},{"Metabolite":"2'-Deoxycytidine","WT1":"1.3E-04","WT2":"9.8E-05","WT3":"9.3E-05","WT4":"2.4E-04","WT5":"N.D.","WT6":"N.D.","KO1":"6.5E-05","KO2":"8.6E-05","KO3":"5.2E-05","KO4":"N.D.","KO5":"N.D.","KO6":"5.8E-05"},{"Metabolite":"3-Amino-2-piperidone","WT1":"5.6E-05","WT2":"N.D.","WT3":"N.D.","WT4":"5.9E-05","WT5":"N.D.","WT6":"N.D.","KO1":"6.8E-05","KO2":"8.6E-05","KO3":"6.6E-05","KO4":"1.3E-04","KO5":"N.D.","KO6":"5.7E-05"},{"Metabolite":"3-Aminobutyric acid","WT1":"5.7E-04","WT2":"9.5E-04","WT3":"4.7E-04","WT4":"8.2E-04","WT5":"8.2E-04","WT6":"9.2E-04","KO1":"2.9E-03","KO2":"1.4E-03","KO3":"9.0E-04","KO4":"1.1E-03","KO5":"1.1E-03","KO6":"1.5E-03"},{"Metabolite":"3-Aminoisobutyric acid","WT1":"5.2E-03","WT2":"3.7E-03","WT3":"3.9E-03","WT4":"7.7E-03","WT5":"1.7E-03","WT6":"3.6E-03","KO1":"4.6E-03","KO2":"5.3E-03","KO3":"2.6E-03","KO4":"2.8E-03","KO5":"5.5E-03","KO6":"5.1E-03"},{"Metabolite":"3-Methylhistidine","WT1":"6.1E-04","WT2":"6.2E-04","WT3":"5.5E-04","WT4":"6.8E-04","WT5":"1.9E-03","WT6":"1.5E-03","KO1":"2.2E-03","KO2":"2.6E-03","KO3":"1.7E-03","KO4":"2.2E-03","KO5":"2.3E-03","KO6":"1.8E-03"},{"Metabolite":"4-Amino-3-hydroxybutyric acid","WT1":"N.D.","WT2":"N.D.","WT3":"N.D.","WT4":"N.D.","WT5":"1.6E-04","WT6":"N.D.","KO1":"1.3E-04","KO2":"2.5E-04","KO3":"1.5E-04","KO4":"1.4E-04","KO5":"1.4E-04","KO6":"1.7E-04"},{"Metabolite":"4-Guanidinobutyric acid","WT1":"8.9E-04","WT2":"1.0E-03","WT3":"2.2E-03","WT4":"2.2E-03","WT5":"6.3E-03","WT6":"4.9E-03","KO1":"4.4E-03","KO2":"1.2E-02","KO3":"9.3E-03","KO4":"1.4E-02","KO5":"7.1E-03","KO6":"6.2E-03"},{"Metabolite":"5-Aminovaleric acid","WT1":"3.6E-04","WT2":"6.7E-04","WT3":"4.1E-04","WT4":"8.5E-04","WT5":"9.5E-03","WT6":"8.2E-03","KO1":"6.1E-03","KO2":"1.4E-02","KO3":"6.9E-03","KO4":"1.1E-02","KO5":"9.1E-03","KO6":"9.8E-03"},{"Metabolite":"5,6,7,8-Tetrahydrobiopterin","WT1":"4.3E-04","WT2":"1.9E-04","WT3":"3.1E-04","WT4":"3.4E-04","WT5":"4.6E-04","WT6":"3.8E-04","KO1":"3.6E-04","KO2":"3.0E-04","KO3":"3.1E-04","KO4":"3.4E-04","KO5":"6.1E-04","KO6":"4.4E-04"},{"Metabolite":"5'-Deoxy-5'-methylthioadenosine","WT1":"6.6E-04","WT2":"6.2E-04","WT3":"4.7E-04","WT4":"5.9E-04","WT5":"5.8E-04","WT6":"4.6E-04","KO1":"5.1E-04","KO2":"4.5E-04","KO3":"2.6E-04","KO4":"4.0E-04","KO5":"4.5E-04","KO6":"3.4E-04"},{"Metabolite":"6-Aminohexanoic 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ID":"C00106","m/z":"113.035","migration time":"21.63"},{"Metabolite":"Urea","KEGG ID":"C00086","m/z":"61.040","migration time":"20.73"},{"Metabolite":"Uridine","KEGG ID":"C00299","m/z":"245.077","migration time":"21.66"},{"Metabolite":"Urocanic acid","KEGG ID":"C00785","m/z":"139.050","migration time":"8.31"},{"Metabolite":"Val","KEGG ID":"C00183,C06417,C16436","m/z":"118.086","migration time":"10.05"},{"Metabolite":"XC0001","KEGG ID":"No ID","m/z":"72.081","migration time":"6.43"},{"Metabolite":"XC0016","KEGG ID":"No ID","m/z":"129.065","migration time":"8.82"},{"Metabolite":"XC0040","KEGG ID":"No ID","m/z":"174.087","migration time":"12.14"},{"Metabolite":"XC0061","KEGG ID":"No ID","m/z":"218.138","migration time":"9.34"},{"Metabolite":"XC0071","KEGG ID":"No ID","m/z":"234.180","migration time":"6.27"},{"Metabolite":"XC0089","KEGG ID":"No ID","m/z":"255.097","migration time":"9.53"},{"Metabolite":"XC0126","KEGG ID":"No ID","m/z":"310.112","migration time":"14.83"},{"Metabolite":"XC0132","KEGG ID":"No ID","m/z":"325.160","migration time":"8.72"},{"Metabolite":"XC0137","KEGG ID":"No ID","m/z":"350.101","migration time":"13.41"},{"Metabolite":"β-Ala","KEGG ID":"C00099","m/z":"90.055","migration time":"7.37"},{"Metabolite":"β-Ala-Lys","KEGG ID":"C05341","m/z":"218.149","migration time":"6.84"},{"Metabolite":"γ-Butyrobetaine","KEGG ID":"C01181","m/z":"146.117","migration time":"8.14"},{"Metabolite":"γ-Glu-2-aminobutyric acid","KEGG ID":"No ID","m/z":"233.112","migration time":"12.47"},{"Metabolite":"γ-Glu-Cys","KEGG ID":"C00669","m/z":"251.069","migration time":"12.75"}]
}

}