#METABOLOMICS WORKBENCH dmw178_20180911_092740_mwtab.txt DATATRACK_ID:1501 STUDY_ID:ST001058 ANALYSIS_ID:AN001734 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 POSITIVE 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 POSITIVE #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 WT_TCP_72H_406 Factors Treatment:No Treatment:No Treatment:No Treatment:No Treatment:No Treatment:No Treatment:Yes Treatment:Yes Treatment:Yes Treatment:Yes Treatment:Yes Treatment:Yes PC 1464710.653 1123030.14 1320976.601 1249467.863 1108022.534 1000888.384 770362.6867 1063668.525 1055449.584 820926.5778 802821.0836 754949.874 PE 6249.1221 3935.0556 6757.0352 5809.2515 9708.2156 9286.1225 2266.0501 3818.8663 2101.3417 5671.6175 2938.6665 8616.603 PG 20.5167 38.3572 33.1627 25.7143 27.632 188.8334 4.2525 57.131 26.8833 40.5 16.0278 20.4055 PI PS 1989.0807 2960.4691 4208.1033 3566.3369 4138.3583 2686.0411 3148.0346 4667.7286 3802.8667 2927.2833 3269.7952 4574.3363 SM 735482.0014 307797.4546 485686.4918 298596.5941 196139.0647 534858.5496 127201.4547 255744.4976 270929.1444 429662.213 146256.3617 281368.9105 Cer 18963.4166 8394.8332 10358.8334 16630.8166 5187.6667 9169.9835 3127.8333 4166.2333 3086.7501 6570.5001 4524.3334 4419.7858 DAG 221 264.7833 203.2 277.3667 219.3333 568.762 185.7667 354.4666 228.4167 941.5334 72.8333 460.3667 TAG 100808.1263 102400.3296 27067.4305 92670.7729 118393.8549 13222.4262 104803.4917 5577.7945 50903.5832 21058.6774 107976.7403 41449.9911 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PC PE PG PI PS SM Cer DAG TAG METABOLITES_END #END