#METABOLOMICS WORKBENCH dmw178_20180911_092740_mwtab.txt DATATRACK_ID:1501 STUDY_ID:ST001058 ANALYSIS_ID:AN001735 PROJECT_ID:PR000708 VERSION 1 CREATED_ON September 24, 2018, 1:12 pm #PROJECT PR:PROJECT_TITLE Metabolic approaches reveal the role of CAR in energy metabolism PR:PROJECT_TYPE Time Course PR:PROJECT_SUMMARY The constitutive androstane receptor (CAR; NR1I3) contributes important PR:PROJECT_SUMMARY regulatory roles in biotransformation, xenobiotic transport function, energy PR:PROJECT_SUMMARY metabolism and lipid homeostasis. In this investigation, global serum and liver PR:PROJECT_SUMMARY tissue metabolomes were assessed analytically in wild type and CAR-null PR:PROJECT_SUMMARY transgenic mice using NMR, GC/MS and UPLC/MS-MS-based metabolomics. PR:PROJECT_SUMMARY Significantly, CAR activation increased serum levels of fatty acids, lactate, PR:PROJECT_SUMMARY ketone bodies and tricarboxylic acid cycle products, whereas levels of PR:PROJECT_SUMMARY phosphatidylcholine, sphingomyelin, amino acids and liver glucose were decreased PR:PROJECT_SUMMARY following short-term activation of CAR. Mechanistically, quantitative mRNA PR:PROJECT_SUMMARY analysis demonstrated significantly decreased expression of key gluconeogenic PR:PROJECT_SUMMARY pathways, and increased expression of glucose utilization pathways, changes PR:PROJECT_SUMMARY likely resulting from down-regulation of the hepatic glucose sensor and PR:PROJECT_SUMMARY bi-directional transporter, Glut2. Short-term CAR activation also resulted in PR:PROJECT_SUMMARY enhanced fatty acid synthesis and impaired β-oxidation. In summary, CAR PR:PROJECT_SUMMARY contributes an expansive role regulating energy metabolism, significantly PR:PROJECT_SUMMARY impacting glucose, and monocarboxylic acid, as well as fatty acid metabolism and PR:PROJECT_SUMMARY lipid homeostasis, through receptor-mediated regulation of several genes in PR:PROJECT_SUMMARY multiple associated pathways. PR:INSTITUTE Pennsylvania State University PR:LABORATORY Omiecinski Lab PR:LAST_NAME Omiecinski PR:FIRST_NAME Curt PR:ADDRESS 101 Life Sciences Building PR:EMAIL cjo10@psu.edu, dmw178@psu.edu PR:PHONE 8148651572 #STUDY ST:STUDY_TITLE Total serum global lipid profiling by UPLC-MS. ST:STUDY_TYPE Time Course ST:STUDY_SUMMARY Liver tissue were harvested from wild type and CAR knockout mice treated for 72h ST:STUDY_SUMMARY with or without TCPOBOP. ST:INSTITUTE Pennsylvania State University ST:LABORATORY Omiecinski Lab ST:LAST_NAME Omiecinski ST:FIRST_NAME Curt ST:ADDRESS 101 Life Sciences Building ST:EMAIL cjo10@psu.edu, dmw178@psu.edu ST:PHONE 8148651572 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 24 ST:NUM_MALES 24 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN Wild Type C57BL/6 and CAR Knockout SU:AGE_OR_AGE_RANGE Approximately 8 weeks old SU:GENDER Male SU:ANIMAL_LIGHT_CYCLE 12 h SU:ANIMAL_FEED ad libitum SU:ANIMAL_WATER ad libitum #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_301 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_302 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_303 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_304 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_305 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_306 Treatment:No TCPOBOP (mg/kg)=-; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_401 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_402 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_403 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_404 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_405 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_406 Treatment:Yes TCPOBOP (mg/kg)=2; Genotype=WT; Time Point (h)=72 #COLLECTION CO:COLLECTION_SUMMARY Blood was harvested via cardiac puncture and then allowed to clot at room CO:COLLECTION_SUMMARY temperature followed by centrifugation. Serum was collected and stored at -80C. CO:SAMPLE_TYPE Blood (serum) CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Each mouse was treated with either a single does of 2 mg/kg of TCPOBOP or the TR:TREATMENT_SUMMARY vehicle control via intraperitoneal injection for 72h. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Serum was prepared using chloroform and methanol extraction. SP:SAMPLEPREP_PROTOCOL_FILENAME LCMS_Lipid_protocol.pdf SP:EXTRACTION_METHOD Methanol-Chloroform SP:SAMPLE_SPIKING Triacetylglyceride (50:1) #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY NEGATIVE CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu Prominence 20 UFLCXR CH:COLUMN_NAME Waters Acquity CSH C18 (100 x 2.1mm, 1.7um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME ABI Sciex 5600+ TripleTOF MS:INSTRUMENT_TYPE Triple TOF MS:MS_TYPE ESI MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak Intensity MS_METABOLITE_DATA_START Samples WT_DMSO_72H_301 WT_DMSO_72H_302 WT_DMSO_72H_303 WT_DMSO_72H_304 WT_DMSO_72H_305 WT_DMSO_72H_306 WT_TCP_72H_401 WT_TCP_72H_402 WT_TCP_72H_403 WT_TCP_72H_404 WT_TCP_72H_405 Factors Treatment:No Treatment:No Treatment:No Treatment:No Treatment:No Treatment:No Treatment:Yes Treatment:Yes Treatment:Yes Treatment:Yes Treatment:Yes PA 619.4 807.0357 2338.8001 199.5833 0 0 32 480.7857 905.4905 173.5 90 PC 31351.2372 16850.4515 38369.6307 17961.1845 6724.0833 9018.4999 11410.7599 25941.3766 31520.1043 12037.025 14382.3517 PE 13359.3741 11122.9487 19292.4976 8333.2805 1276.5833 3564.75 8920.85 10686.6484 12678.1286 11916.5784 4579.981 PG 6072.2083 3672.5333 11004.1007 621.9167 57.25 323.3334 7688.75 22246.4667 1275.8584 4465.5834 832.3334 PI 8340.9402 7467.6612 17851.8708 7727.9777 1164 1989.6667 4780.35 8723.7501 10134.8848 5594.6334 4042.6666 PS 13128.2655 18660.6379 27329.5806 5715.7737 1679.5834 3565.3055 11716.919 14738.7386 20646.0222 14262.4461 9847.9033 SM 7351.1667 8234.6667 8975.4167 4918.8333 870 1988 3347 5171.8 6307.8332 5973.3334 3287 FFA 7675.4916 11956.3816 15979.0795 9982.7392 2706.25 11644.4641 10218.5918 16154.6238 10952.5716 7426.0596 7425.6583 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PA PC PE PG PI PS SM FFA METABOLITES_END #END