#METABOLOMICS WORKBENCH dmw178_20180911_072635 DATATRACK_ID:1499 STUDY_ID:ST001092 ANALYSIS_ID:AN001778 PROJECT_ID:PR000708 VERSION 1 CREATED_ON November 6, 2018, 4:06 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 Metabolic profiling by NMR analysis in liver aqueous extract samples ST:STUDY_TYPE Time Course ST:STUDY_SUMMARY Liver tissue were harvested from wild type and CAR knockout mice treated for 48 ST:STUDY_SUMMARY or 72h 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 8 ST:TOTAL_SUBJECTS 48 ST:NUM_MALES 48 #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_48H_101 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_48H_102 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_48H_103 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_48H_104 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_48H_105 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_48H_106 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_201 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_202 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_203 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_204 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_205 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_48H_206 Genotype:WT | Time Point (h):48 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_301 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_302 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_303 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_304 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_305 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_DMSO_72H_306 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):- SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_401 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_402 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_403 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_404 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_405 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 SUBJECT_SAMPLE_FACTORS - WT_TCP_72H_406 Genotype:WT | Time Point (h):72 | TCPOBOP (mg/kg):2 #COLLECTION CO:COLLECTION_SUMMARY Liver tissue was snap-frozen in liquid nitrogen and stored at -80C. CO:SAMPLE_TYPE Liver CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Each mouse was treated with either a singel does of 2 mg/kg of Car agonist TR:TREATMENT_SUMMARY (TCPOBOP) or the vehicle control via intraperitoneal injection. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Liver tissue was homogenized in a methanol-water mixture, dried and resuspended SP:SAMPLEPREP_SUMMARY in phosphate buffer containing TSP-d4. SP:SAMPLEPREP_PROTOCOL_FILENAME NMR_metabolomics_protocol.pdf SP:PROCESSING_METHOD Homogenization SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACTION_METHOD Methanol/Water SP:EXTRACT_STORAGE On ice SP:SAMPLE_RESUSPENSION D2O SP:SAMPLE_SPIKING TSP-d4 #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE - CH:INSTRUMENT_NAME - CH:COLUMN_NAME - #ANALYSIS AN:ANALYSIS_TYPE NMR AN:LABORATORY_NAME Omiecinski Lab #NMR NM:INSTRUMENT_NAME Bruker Avance III NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE Other NM:NMR_COMMENTS NOESYPR1D NM:SPECTROMETER_FREQUENCY 600 MHz NM:NMR_SOLVENT D2O NM:NMR_RESULTS_FILE ST001092_AN001778_Results.txt UNITS:ppm #END