{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001622","ANALYSIS_ID":"AN002657","VERSION":"1","CREATED_ON":"December 11, 2020, 10:20 am"},

"PROJECT":{"PROJECT_TITLE":"Isotope Tracing Analysis in Intestinal Organoids","PROJECT_TYPE":"Isotope Tracing Analysis","PROJECT_SUMMARY":"To elucidate the mechanism by which increased fructose production favors ISC proliferation, We performed untargeted metabolomics of intestinal organoids treated with fructose (10 mM) for 24 hr. As expected,Typically, metabolites such as glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), lactate, pyruvate, citrate and glutamine were markedly enriched after fructose treatment.To gain more functional insights, we further conducted isotope tracing analysis of U-[13C]-fructose in intestinal organoids. [13C] tracing revealed that fructose-treated organoids had higher percentage of fully labeled F1P(M+6),lactate (M+3),G6P(M+6), F6P(M+6), supporting the enhancement of glycolysis after fructose treatment Together, these results indicate that fructose induces a metabolic reprogramming in intestinal organoids that favors glycolysis.","INSTITUTE":"China Pharmaceutical University","DEPARTMENT":"School of Medicine","LABORATORY":"Metabonomics","LAST_NAME":"Hou","FIRST_NAME":"Yuanlong","ADDRESS":"tongjiaxiang, nanjing, jiangsu, 210000, China","EMAIL":"jian2103@163.com","PHONE":"18851105337","CONTRIBUTORS":"Xiao Zheng Haiping Hao"},

"STUDY":{"STUDY_TITLE":"Isotope Tracing Analysis in Intestinal Organoids with fructose","STUDY_SUMMARY":"We isolated and cultured Intestinal Organoids from Mouse. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr.","INSTITUTE":"China Pharmaceutical University","DEPARTMENT":"School of Medicine","LABORATORY":"Metabonomics","LAST_NAME":"Hou","FIRST_NAME":"Yuanlong","ADDRESS":"tongjiaxiang, nanjing, jiangsu, 210000, China","EMAIL":"jian2103@163.com","PHONE":"18851105337"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENDER":"Female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"13C Fru1",
"Factors":{"Treatment":"13C Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-c13 FRUCTOSE1"}
},
{
"Subject ID":"-",
"Sample ID":"13C Fru2",
"Factors":{"Treatment":"13C Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-c13 FRUCTOSE2"}
},
{
"Subject ID":"-",
"Sample ID":"13C Fru3",
"Factors":{"Treatment":"13C Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-c13 FRUCTOSE3"}
},
{
"Subject ID":"-",
"Sample ID":"Fru1",
"Factors":{"Treatment":"Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-FRUCTOSE1"}
},
{
"Subject ID":"-",
"Sample ID":"Fru2",
"Factors":{"Treatment":"Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-FRUCTOSE2"}
},
{
"Subject ID":"-",
"Sample ID":"Fru3",
"Factors":{"Treatment":"Fructose"},
"Additional sample data":{"RAW_FILE_NAME":"20200416-FRUCTOSE3"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Isolation and Culture of Mouse Intestinal Organoids","SAMPLE_TYPE":"Intestine"},

"TREATMENT":{"TREATMENT_SUMMARY":"For metabolic flux analysis of 13C-labeled glucose, intestinal organoids were pretreated with unlabeled fructose (10 mM) for 24 hr, washed with PBS, and then switched into 13C labeled glucose-containing DMEM/F12 medium (10 mM U-[13C] D-glucose, Cambridge Isotope Laboratories) for 6 hr. To trace the metabolism of fructose, intestinal organoids were cultured in unlabeled fructose-containing DMEM/F12 medium (10 mM D-fructose, 0 mM D-glucose, 2.5 mM L-glutamine) for 12 hr, and then switched into 13C labeled fructose-containing medium (10 mM U-[13C] D-fructose, Sigma-aldrich) for 6 hr."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"To harvest intracellular metabolites, organoid samples were washed with pre-cooled PBS for three times and metabolism was quenched with 80% ice-cold methanol. Pre-cooled methanol (1mL) with para-chlorophenylalanine (1 µM) as the internal standard was added to each sample and incubated on ice for 10 min. Sample lysates were then centrifuged at 4 °C for 15 min at 23,000 g."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"UPLC-Q-TOF-MS (Synapt G2si, waters)","COLUMN_NAME":"Waters XBridge Amide (100 x 4.6mm, 3.5um)"},

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

"MS":{"INSTRUMENT_NAME":"Waters Synapt G2 Si QTOF","INSTRUMENT_TYPE":"GC-TOF","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"MS MassLynx v4.1 Quanlynx v4.1"},

"MS_METABOLITE_DATA":{
"Units":"abundance",

"Data":[{"Metabolite":"Fructose 6-Phosphate(M+0)","Fru1":"318734","Fru2":"293265","Fru3":"270236","13C Fru1":"96537","13C Fru2":"97564","13C Fru3":"108955"},{"Metabolite":"Fructose 6-Phosphate(M+1)","Fru1":"14012","Fru2":"12463","Fru3":"12974","13C Fru1":"8324","13C Fru2":"8304","13C Fru3":"9283"},{"Metabolite":"Fructose 6-Phosphate(M+2)","Fru1":"4852","Fru2":"4624","Fru3":"4007","13C Fru1":"6197","13C Fru2":"6261","13C Fru3":"5971"},{"Metabolite":"Fructose 6-Phosphate(M+3)","Fru1":"250","Fru2":"345","Fru3":"497","13C Fru1":"11197","13C Fru2":"12981","13C Fru3":"13175"},{"Metabolite":"Fructose 6-Phosphate(M+4)","Fru1":"184","Fru2":"214","Fru3":"131","13C Fru1":"7213","13C Fru2":"7846","13C Fru3":"7989"},{"Metabolite":"Fructose 6-Phosphate(M+5)","Fru1":"82","Fru2":"79","Fru3":"141","13C Fru1":"12276","13C Fru2":"14890","13C Fru3":"12295"},{"Metabolite":"Fructose 6-Phosphate(M+6)","Fru1":"100","Fru2":"100","Fru3":"100","13C Fru1":"77189","13C Fru2":"108767","13C Fru3":"80903"},{"Metabolite":"Glucose 6-Phosphate(M+0)","Fru1":"367399","Fru2":"389864","Fru3":"431494","13C Fru1":"148580","13C Fru2":"96667","13C Fru3":"142538"},{"Metabolite":"Glucose 6-Phosphate(M+1)","Fru1":"17803","Fru2":"17465","Fru3":"19004","13C Fru1":"12800","13C Fru2":"8929","13C Fru3":"12234"},{"Metabolite":"Glucose 6-Phosphate(M+2)","Fru1":"5325","Fru2":"5959","Fru3":"6558","13C Fru1":"8355","13C Fru2":"8137","13C Fru3":"6250"},{"Metabolite":"Glucose 6-Phosphate(M+3)","Fru1":"632","Fru2":"424","Fru3":"492","13C Fru1":"14609","13C Fru2":"13634","13C Fru3":"14043"},{"Metabolite":"Glucose 6-Phosphate(M+4)","Fru1":"230","Fru2":"287","Fru3":"211","13C Fru1":"9166","13C Fru2":"8061","13C Fru3":"8867"},{"Metabolite":"Glucose 6-Phosphate(M+5)","Fru1":"188","Fru2":"113","Fru3":"139","13C Fru1":"16590","13C Fru2":"18441","13C Fru3":"14581"},{"Metabolite":"Glucose 6-Phosphate(M+6)","Fru1":"388","Fru2":"259","Fru3":"308","13C Fru1":"126708","13C Fru2":"180706","13C Fru3":"91999"},{"Metabolite":"Fructose 1-Phosphate(M+0)","Fru1":"115938","Fru2":"94650","Fru3":"63186","13C Fru1":"85723","13C Fru2":"78094","13C Fru3":"71650"},{"Metabolite":"Fructose 1-Phosphate(M+1)","Fru1":"5398","Fru2":"4984","Fru3":"3630","13C Fru1":"5140","13C Fru2":"5397","13C Fru3":"4815"},{"Metabolite":"Fructose 1-Phosphate(M+2)","Fru1":"1089","Fru2":"1602","Fru3":"1977","13C Fru1":"2281","13C Fru2":"2416","13C Fru3":"1714"},{"Metabolite":"Fructose 1-Phosphate(M+3)","Fru1":"10","Fru2":"10","Fru3":"10","13C Fru1":"1924","13C Fru2":"3047","13C Fru3":"2389"},{"Metabolite":"Fructose 1-Phosphate(M+4)","Fru1":"130","Fru2":"20","Fru3":"10","13C Fru1":"1571","13C Fru2":"1866","13C Fru3":"1246"},{"Metabolite":"Fructose 1-Phosphate(M+5)","Fru1":"132","Fru2":"20","Fru3":"10","13C Fru1":"3028","13C Fru2":"3650","13C Fru3":"2770"},{"Metabolite":"Fructose 1-Phosphate(M+6)","Fru1":"169","Fru2":"220","Fru3":"100","13C Fru1":"28444","13C Fru2":"26499","13C Fru3":"33696"},{"Metabolite":"Lactate(M+0)","Fru1":"406188","Fru2":"396546","Fru3":"102658","13C Fru1":"102587","13C Fru2":"128709","13C Fru3":"129655"},{"Metabolite":"Lactate(M+1)","Fru1":"10006","Fru2":"9557","Fru3":"3952","13C Fru1":"7312","13C Fru2":"10131","13C Fru3":"8409"},{"Metabolite":"Lactate(M+2)","Fru1":"2868","Fru2":"2890","Fru3":"1552","13C Fru1":"10014","13C Fru2":"13233","13C Fru3":"11207"},{"Metabolite":"Lactate(M+3)","Fru1":"10","Fru2":"10","Fru3":"10","13C Fru1":"224952","13C Fru2":"313760","13C Fru3":"264830"}],

"Metabolites":[{"Metabolite":"Fructose 6-Phosphate(M+0)","retention index":"8.82","quantified m/z":"259.0219","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+1)","retention index":"8.82","quantified m/z":"260.0252","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+2)","retention index":"8.82","quantified m/z":"261.0286","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+3)","retention index":"8.82","quantified m/z":"262.032","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+4)","retention index":"8.82","quantified m/z":"263.0353","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+5)","retention index":"8.82","quantified m/z":"264.0387","HMDB ID":"HMDB0000124"},{"Metabolite":"Fructose 6-Phosphate(M+6)","retention index":"8.82","quantified m/z":"265.042","HMDB ID":"HMDB0000124"},{"Metabolite":"Glucose 6-Phosphate(M+0)","retention index":"8.78","quantified m/z":"259.0219","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+1)","retention index":"8.78","quantified m/z":"260.0252","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+2)","retention index":"8.78","quantified m/z":"261.0286","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+3)","retention index":"8.78","quantified m/z":"262.032","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+4)","retention index":"8.78","quantified m/z":"263.0353","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+5)","retention index":"8.78","quantified m/z":"264.0387","HMDB ID":"HMDB0001401"},{"Metabolite":"Glucose 6-Phosphate(M+6)","retention index":"8.78","quantified m/z":"265.042","HMDB ID":"HMDB0001401"},{"Metabolite":"Fructose 1-Phosphate(M+0)","retention index":"8.7","quantified m/z":"259.0219","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+1)","retention index":"8.7","quantified m/z":"260.0252","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+2)","retention index":"8.7","quantified m/z":"261.0286","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+3)","retention index":"8.7","quantified m/z":"262.032","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+4)","retention index":"8.7","quantified m/z":"263.0353","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+5)","retention index":"8.7","quantified m/z":"264.0387","HMDB ID":"HMDB0001076"},{"Metabolite":"Fructose 1-Phosphate(M+6)","retention index":"8.7","quantified m/z":"265.042","HMDB ID":"HMDB0001076"},{"Metabolite":"Lactate(M+0)","retention index":"6.73","quantified m/z":"89.0239","HMDB ID":"HMDB0000190"},{"Metabolite":"Lactate(M+1)","retention index":"6.73","quantified m/z":"90.0272","HMDB ID":"HMDB0000190"},{"Metabolite":"Lactate(M+2)","retention index":"6.73","quantified m/z":"91.0306","HMDB ID":"HMDB0000190"},{"Metabolite":"Lactate(M+3)","retention index":"6.73","quantified m/z":"92.0339","HMDB ID":"HMDB0000190"}]
}

}