{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003300","ANALYSIS_ID":"AN005407","PROJECT_ID":"PR002050","VERSION":"1","CREATED_ON":"May 13, 2024, 9:17 pm"},

"PROJECT":{"PROJECT_TITLE":"Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue","PROJECT_TYPE":"Mass spectrometry","PROJECT_SUMMARY":"The central nervous system has been implicated in the age-induced reduction in adipose tissue lipolysis. SLC7A14 is a lysosomal membrane protein highly expressed in the brain. Herein, we investigated the possible role of hypothalamic SLC7A14 in the age-induced lipolysis reduction. In this study, we demonstrated the expression of SLC7A14 was reduced in proopiomelanocortin (POMC) neurons of aged mice. Overexpression of SLC7A14 in POMC neurons alleviated the age-induced reduction in white adipose tissue (WAT) lipolysis, whereas SLC7A14 deletion mimicked the age-induced lipolysis impairment. Moreover, POMC SLC7A14 regulated WAT lipolysis independently of sympathetic nerves in WAT. Metabolomics analysis revealed that POMC SLC7A14 increased the primary bile acid taurochenodeoxycholic acid (TCDCA) content, which mediated the SLC7A14 knockout- or age-induced WAT lipolysis impairment. Furthermore, SLC7A14-increased TCDCA content is dependent on intestinal apical sodium-dependent bile acid transporter (ASBT), which is regulated by intestinal sympathetic afferent nerves. Finally, SLC7A14 regulated the intestinal sympathetic afferent nerves by inhibiting mTORC1 signaling through inhibiting TSC1 phosphorylation. Collectively, our study suggests the function for central SLC7A14 and an upstream mechanism for the mTORC1 signaling pathway. Moreover, our data provides insights into the brain–gut–adipose tissue crosstalk in age-induced lipolysis impairment.","INSTITUTE":"Shanghai Institutes for Biological Sciences (SIBS) Chinese Academy of Sciences (CAS)","LAST_NAME":"Liu","FIRST_NAME":"Kan","ADDRESS":"No. 320, Yueyang Road, Shanghai, Shanghai, Shanghai/Shanghai/xuhui, 200000, China","EMAIL":"liukan2019@sibs.ac.cn","PHONE":"021-17718134725"},

"STUDY":{"STUDY_TITLE":"Hypothalamic SLC7A14 accounts for aging-reduced lipolysis in white adipose tissue","STUDY_SUMMARY":"The central nervous system has been implicated in the age-induced reduction in adipose tissue lipolysis. SLC7A14 is a lysosomal membrane protein highly expressed in the brain. Herein, we investigated the possible role of hypothalamic SLC7A14 in the age-induced lipolysis reduction. In this study, we demonstrated the expression of SLC7A14 was reduced in proopiomelanocortin (POMC) neurons of aged mice. Overexpression of SLC7A14 in POMC neurons alleviated the age-induced reduction in white adipose tissue (WAT) lipolysis, whereas SLC7A14 deletion mimicked the age-induced lipolysis impairment. Moreover, POMC SLC7A14 regulated WAT lipolysis independently of sympathetic nerves in WAT. Metabolomics analysis revealed that POMC SLC7A14 increased the primary bile acid taurochenodeoxycholic acid (TCDCA) content, which mediated the SLC7A14 knockout- or age-induced WAT lipolysis impairment. Furthermore, SLC7A14-increased TCDCA content is dependent on intestinal apical sodium-dependent bile acid transporter (ASBT), which is regulated by intestinal sympathetic afferent nerves. Finally, SLC7A14 regulated the intestinal sympathetic afferent nerves by inhibiting mTORC1 signaling through inhibiting TSC1 phosphorylation. Collectively, our study suggests the function for central SLC7A14 and an upstream mechanism for the mTORC1 signaling pathway. Moreover, our data provides insights into the brain–gut–adipose tissue crosstalk in age-induced lipolysis impairment.","INSTITUTE":"Shanghai Institutes for Biological Sciences (SIBS) Chinese Academy of Sciences (CAS)","LAST_NAME":"Liu","FIRST_NAME":"Kan","ADDRESS":"No. 320, Yueyang Road, Shanghai","EMAIL":"liukan2019@sibs.ac.cn","PHONE":"021-17718134725"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090","GENDER":"Male"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"control3",
"Factors":{"Sample source":"Blood serum","Genotype":"WT","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN1_FZTM210013175-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"control1",
"Factors":{"Sample source":"Blood serum","Genotype":"WT","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN1_FZTM210013176-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"control2",
"Factors":{"Sample source":"Blood serum","Genotype":"WT","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN1_FZTM210013174-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"control4",
"Factors":{"Sample source":"Blood serum","Genotype":"WT","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN1_FZTM210013177-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"control5",
"Factors":{"Sample source":"Blood serum","Genotype":"WT","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN1_FZTM210013178-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"oe1",
"Factors":{"Sample source":"Blood serum","Genotype":"SLC7A14_OE","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN2_FZTM210013180-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"oe2",
"Factors":{"Sample source":"Blood serum","Genotype":"SLC7A15_OE","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN2_FZTM210013181-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"oe3",
"Factors":{"Sample source":"Blood serum","Genotype":"SLC7A16_OE","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN2_FZTM210013182-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"oe4",
"Factors":{"Sample source":"Blood serum","Genotype":"SLC7A17_OE","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_CN2_FZTM210013183-1A.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"QC1",
"Factors":{"Sample source":"Blood serum","Genotype":"QC","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_663032_CN_QC1.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"QC2",
"Factors":{"Sample source":"Blood serum","Genotype":"QC","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_663032_CN_QC2.mzML"}
},
{
"Subject ID":"-",
"Sample ID":"QC3",
"Factors":{"Sample source":"Blood serum","Genotype":"QC","Treatment":"control"},
"Additional sample data":{"RAW_FILE_NAME(Raw file name)":"HFX5_663032_CN_QC3.mzML"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice.","SAMPLE_TYPE":"Blood (serum)"},

"TREATMENT":{"TREATMENT_SUMMARY":"To overexpression of SLC7A14 in ARC POMC neurons, POMC Cre mice were bilaterally injected either with a Cre-dependent AAV vector containing SLC7A14 in the opposite orientation flanked by two inverted loxP sites (AAV9-Syn-DIO-SLC7A14-mCherry, 1.5 × 1012 Pfu/mL, HANBIO) at a volume of 200 nL into the ARC or an AAV vector containing only mCherry in the opposite orientation flanked by two inverted loxP sites (AAV9-Syn-DIO-mCherry, 1.5 × 1012 Pfu/mL, HANBIO) as a control.Serum was collected from control or overexpression of SLC7A14 in POMC neuron mice."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"The samples (100 μL) were placed in the EP tubes and resuspended with prechilled 80% methanol and 0.1% formic acid by well vortex. Then the sampleswere incubated on ice for 5 min and centrifuged at 15,000 g, 4°C for 20 min. Some of supernatant was diluted to final concentration containing 53% methanol by LC-MS grade water.The samples were subsequently transferred to a fresh Eppendorf tube and then were centrifuged at 15000 g, 4°C for 20 min. Finally, the supernatant was injected into the LC-MS/MS system analysis"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Thermo Hypersil GOLD aQ (100 x 2.1mm,1.9um)","SOLVENT_A":"The eluents for the negative polarity mode were eluent A (5 mMammonium acetate, pH 9.0","SOLVENT_B":"Methanol","FLOW_GRADIENT":"2% B, 1.5 min; 2-100% B, 12.0 min; 100% B, 14.0 min；100-2% B, 14.1 min；2% B, 17 min.","FLOW_RATE":"0.2 mL/min","COLUMN_TEMPERATURE":"40"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive HF-X Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"NEGATIVE","MS_COMMENTS":"The raw data files generated by UHPLC-MS/MS were processed using the Compound Discoverer 3.1 (CD3.1, ThermoFisher) to perform peak alignment, peak picking, and quantitation for each metabolite. The main parameterswere set as follows: retention time tolerance, 0.2 minutes; actual mass tolerance, 5ppm; signal intensity tolerance, 30%; signal/noise ratio, 3; and minimum intensity, et al. After that, peak intensities were normalized to the total spectral intensity.The normalized data was used to predict the molecular formula based on additive ions, molecular ion peaks and fragment ions. And then peaks were matched with the mzCloud (https://www.mzcloud.org/)，mzVault and MassList database to obtain the accurate qualitative and relative quantitative results.Statistical analyses were performed using the statistical software R (R version R-3.4.3),Python (Python 2.7.6 version) and CentOS (CentOS release 6.6),When data were not normally distributed, normal transformations were attempted using of area normalization method.","MS_RESULTS_FILE":"ST003300_AN005407_Results.txt UNITS:Peak area Has m/z:Yes Has RT:Yes RT units:Minutes"}

}