#METABOLOMICS WORKBENCH maxmendez_20210627_091843 DATATRACK_ID:2720 STUDY_ID:ST001858 ANALYSIS_ID:AN003012 PROJECT_ID:000000 VERSION 1 CREATED_ON June 28, 2021, 1:00 pm #PROJECT PR:PROJECT_TITLE Proteasome Inhibitor Related Cardiotoxicity PR:PROJECT_TYPE MS Untargeted Metabolomics from Murine Plasma PR:PROJECT_SUMMARY In this study we aimed to examine the acute metabolomic changes following PR:PROJECT_SUMMARY proteasome inhibition with Bortezomib or Carfilzomib in murine plasma. PR:INSTITUTE University of Zurich PR:DEPARTMENT Department of Hematology-Oncology. Kantonsspital St. Gallen PR:LABORATORY Experimental Hematolgy-KSSG PR:LAST_NAME Mendez PR:FIRST_NAME Max PR:ADDRESS Rorschacherstrasse 95, 9007, St. Gallen. PR:EMAIL maxmendez@runbox.com PR:PHONE +41 71 494 10 67 #STUDY ST:STUDY_TITLE Proteasome Inhibitor Related Cardiotoxicity (part II) ST:STUDY_SUMMARY BALB/c mice were injected with CFZ (16 mg/kg, iv), BTZ (1 mg/kg, iv), CFZ/atRA ST:STUDY_SUMMARY (CFZ: 16 mg/kg iv, atRA: 1 mg/kg iv) or Captisol (Vehicle, iv) (6 mice per ST:STUDY_SUMMARY cohort). ECGs were recorded under anesthesia at the end of 1h treatment and the ST:STUDY_SUMMARY mice were sacrificed by cervical dislocation. Plasma was obtained after ST:STUDY_SUMMARY centrifugation by heart puncture. Samples were immediately frozen and stored at ST:STUDY_SUMMARY -80°C until analysis ST:INSTITUTE University of Zurich ST:LAST_NAME Mendez ST:FIRST_NAME Max ST:ADDRESS Rorschacherstrasse 95. 9007, St.Gallen ST:EMAIL maxmendez@runbox.com ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 24 ST:NUM_FEMALES 24 ST:PHONE +41 71 494 10 67 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN Balb/c SU:AGE_OR_AGE_RANGE 8-12 weeks old SU:WEIGHT_OR_WEIGHT_RANGE 20-25 g SU:GENDER Female 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]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - QC_06_NEG class:QC RAW_FILE_NAME=QC_06_NEG; order=1 SUBJECT_SAMPLE_FACTORS - QC_07_NEG class:QC RAW_FILE_NAME=QC_07_NEG; order=2 SUBJECT_SAMPLE_FACTORS - QC_08_NEG class:QC RAW_FILE_NAME=QC_08_NEG; order=3 SUBJECT_SAMPLE_FACTORS - QC_09_NEG class:QC RAW_FILE_NAME=QC_09_NEG; order=4 SUBJECT_SAMPLE_FACTORS - QC_10_NEG class:QC RAW_FILE_NAME=QC_10_NEG; order=5 SUBJECT_SAMPLE_FACTORS - pCTRL1_NEG class:Pcontrol RAW_FILE_NAME=pCTRL1_NEG; order=6 SUBJECT_SAMPLE_FACTORS - pCFZ1_NEG class:PcaseB RAW_FILE_NAME=pCFZ1_NEG; order=7 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA1_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA1_NEG; order=8 SUBJECT_SAMPLE_FACTORS - pBTZ1_NEG class:PcaseD RAW_FILE_NAME=pBTZ1_NEG; order=9 SUBJECT_SAMPLE_FACTORS - pCTRL2_NEG class:Pcontrol RAW_FILE_NAME=pCTRL2_NEG; order=10 SUBJECT_SAMPLE_FACTORS - pCFZ2_NEG class:PcaseB RAW_FILE_NAME=pCFZ2_NEG; order=11 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA2_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA2_NEG; order=12 SUBJECT_SAMPLE_FACTORS - pBTZ2_NEG class:PcaseD RAW_FILE_NAME=pBTZ2_NEG; order=13 SUBJECT_SAMPLE_FACTORS - pCTRL3_NEG class:Pcontrol RAW_FILE_NAME=pCTRL3_NEG; order=14 SUBJECT_SAMPLE_FACTORS - pCFZ3_NEG class:PcaseB RAW_FILE_NAME=pCFZ3_NEG; order=15 SUBJECT_SAMPLE_FACTORS - QC_11_NEG class:QC RAW_FILE_NAME=QC_11_NEG; order=16 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA3_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA3_NEG; order=17 SUBJECT_SAMPLE_FACTORS - pBTZ3_NEG class:PcaseD RAW_FILE_NAME=pBTZ3_NEG; order=18 SUBJECT_SAMPLE_FACTORS - pCTRL4_NEG class:Pcontrol RAW_FILE_NAME=pCTRL4_NEG; order=19 SUBJECT_SAMPLE_FACTORS - pCFZ4_NEG class:PcaseB RAW_FILE_NAME=pCFZ4_NEG; order=20 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA4_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA4_NEG; order=21 SUBJECT_SAMPLE_FACTORS - pBTZ4_NEG class:PcaseD RAW_FILE_NAME=pBTZ4_NEG; order=22 SUBJECT_SAMPLE_FACTORS - pCTRL5_NEG class:Pcontrol RAW_FILE_NAME=pCTRL5_NEG; order=23 SUBJECT_SAMPLE_FACTORS - pCFZ5_NEG class:PcaseB RAW_FILE_NAME=pCFZ5_NEG; order=24 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA5_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA5_NEG; order=25 SUBJECT_SAMPLE_FACTORS - pBTZ5_NEG class:PcaseD RAW_FILE_NAME=pBTZ5_NEG; order=26 SUBJECT_SAMPLE_FACTORS - QC_12_NEG class:QC RAW_FILE_NAME=QC_12_NEG; order=27 SUBJECT_SAMPLE_FACTORS - pCTRL6_NEG class:Pcontrol RAW_FILE_NAME=pCTRL6_NEG; order=28 SUBJECT_SAMPLE_FACTORS - pCFZ6_NEG class:PcaseB RAW_FILE_NAME=pCFZ6_NEG; order=29 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA6_NEG class:PcaseC RAW_FILE_NAME=pCFZ.ATRA6_NEG; order=30 SUBJECT_SAMPLE_FACTORS - pBTZ6_NEG class:PcaseD RAW_FILE_NAME=pBTZ6_NEG; order=31 SUBJECT_SAMPLE_FACTORS - QC_13_NEG class:QC RAW_FILE_NAME=QC_13_NEG; order=32 SUBJECT_SAMPLE_FACTORS - QC_14_NEG class:QC RAW_FILE_NAME=QC_14_NEG; order=33 SUBJECT_SAMPLE_FACTORS - QC_15_NEG class:QC RAW_FILE_NAME=QC_15_NEG; order=34 SUBJECT_SAMPLE_FACTORS - QC_06_POS class:QC RAW_FILE_NAME=QC_06_POS; order=1 SUBJECT_SAMPLE_FACTORS - QC_07_POS class:QC RAW_FILE_NAME=QC_07_POS; order=2 SUBJECT_SAMPLE_FACTORS - QC_08_POS class:QC RAW_FILE_NAME=QC_08_POS; order=3 SUBJECT_SAMPLE_FACTORS - QC_09_POS class:QC RAW_FILE_NAME=QC_09_POS; order=4 SUBJECT_SAMPLE_FACTORS - QC_10_POS class:QC RAW_FILE_NAME=QC_10_POS; order=5 SUBJECT_SAMPLE_FACTORS - pCTRL1_POS class:Pcontrol RAW_FILE_NAME=pCTRL1_POS; order=6 SUBJECT_SAMPLE_FACTORS - pCFZ1_POS class:PcaseB RAW_FILE_NAME=pCFZ1_POS; order=7 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA1_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA1_POS; order=8 SUBJECT_SAMPLE_FACTORS - pBTZ1_POS class:PcaseD RAW_FILE_NAME=pBTZ1_POS; order=9 SUBJECT_SAMPLE_FACTORS - pCTRL2_POS class:Pcontrol RAW_FILE_NAME=pCTRL2_POS; order=10 SUBJECT_SAMPLE_FACTORS - pCFZ2_POS class:PcaseB RAW_FILE_NAME=pCFZ2_POS; order=11 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA2_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA2_POS; order=12 SUBJECT_SAMPLE_FACTORS - pBTZ2_POS class:PcaseD RAW_FILE_NAME=pBTZ2_POS; order=13 SUBJECT_SAMPLE_FACTORS - pCTRL3_POS class:Pcontrol RAW_FILE_NAME=pCTRL3_POS; order=14 SUBJECT_SAMPLE_FACTORS - pCFZ3_POS class:PcaseB RAW_FILE_NAME=pCFZ3_POS; order=15 SUBJECT_SAMPLE_FACTORS - QC_11_POS class:QC RAW_FILE_NAME=QC_11_POS; order=16 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA3_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA3_POS; order=17 SUBJECT_SAMPLE_FACTORS - pBTZ3_POS class:PcaseD RAW_FILE_NAME=pBTZ3_POS; order=18 SUBJECT_SAMPLE_FACTORS - pCTRL4_POS class:Pcontrol RAW_FILE_NAME=pCTRL4_POS; order=19 SUBJECT_SAMPLE_FACTORS - pCFZ4_POS class:PcaseB RAW_FILE_NAME=pCFZ4_POS; order=20 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA4_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA4_POS; order=21 SUBJECT_SAMPLE_FACTORS - pBTZ4_POS class:PcaseD RAW_FILE_NAME=pBTZ4_POS; order=22 SUBJECT_SAMPLE_FACTORS - pCTRL5_POS class:Pcontrol RAW_FILE_NAME=pCTRL5_POS; order=23 SUBJECT_SAMPLE_FACTORS - pCFZ5_POS class:PcaseB RAW_FILE_NAME=pCFZ5_POS; order=24 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA5_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA5_POS; order=25 SUBJECT_SAMPLE_FACTORS - pBTZ5_POS class:PcaseD RAW_FILE_NAME=pBTZ5_POS; order=26 SUBJECT_SAMPLE_FACTORS - QC_12_POS class:QC RAW_FILE_NAME=QC_12_POS; order=27 SUBJECT_SAMPLE_FACTORS - pCTRL6_POS class:Pcontrol RAW_FILE_NAME=pCTRL6_POS; order=28 SUBJECT_SAMPLE_FACTORS - pCFZ6_POS class:PcaseB RAW_FILE_NAME=pCFZ6_POS; order=29 SUBJECT_SAMPLE_FACTORS - pCFZ.ATRA6_POS class:PcaseC RAW_FILE_NAME=pCFZ.ATRA6_POS; order=30 SUBJECT_SAMPLE_FACTORS - pBTZ6_POS class:PcaseD RAW_FILE_NAME=pBTZ6_POS; order=31 SUBJECT_SAMPLE_FACTORS - QC_13_POS class:QC RAW_FILE_NAME=QC_13_POS; order=32 SUBJECT_SAMPLE_FACTORS - QC_14_POS class:QC RAW_FILE_NAME=QC_14_POS; order=33 SUBJECT_SAMPLE_FACTORS - QC_15_POS class:QC RAW_FILE_NAME=QC_15_POS; order=34 #COLLECTION CO:COLLECTION_SUMMARY Anesthesia was induced with Ketamine 80 mg/kg intramuscular and maintained with CO:COLLECTION_SUMMARY Isoflurane 0.5%. Plasma was obtained by centrifugation after cervical CO:COLLECTION_SUMMARY dislocation under anesthesia. Samples were snap frozen in dry ice and kept at CO:COLLECTION_SUMMARY -80°C until analysis. CO:SAMPLE_TYPE Blood (plasma) CO:STORAGE_CONDITIONS Described in summary #TREATMENT TR:TREATMENT_SUMMARY Injections with CFZ (16 mg/kg, iv), BTZ (1 mg/kg, iv), CFZ+atRA (CFZ: 16 mg/kg TR:TREATMENT_SUMMARY iv, atRA: 1 mg/kg iv) or Captisol (Vehicle, iv) TR:TREATMENT_ROUTE i.v TR:TREATMENT_DOSE As described in treatment summary TR:TREATMENT_VEHICLE Captisol TR:ANIMAL_ANESTHESIA Induction with Ketamine 80 mg/kg intramuscular and maintained with Isoflurane TR:ANIMAL_ANESTHESIA 0.5% TR:ANIMAL_ENDP_EUTHANASIA 1h TR:ANIMAL_ENDP_TISSUE_COLL_LIST 1h TR:TREATMENT_COMPOUND Bortezomib (PS-34, Selleck Chemicals, S1013); Carfilzomib (PR-171, Selleck TR:TREATMENT_COMPOUND Chemicals, S2853); all-trans-Retinoic-Acid (Sigma-Aldrich, R2625); Captisol TR:TREATMENT_COMPOUND (Ligand Pharmaceuticals #SAMPLEPREP SP:SAMPLEPREP_SUMMARY 1 Take 100 μL of each sample (including QC) into the corresponding 96-well SP:SAMPLEPREP_SUMMARY plate; 2 Add 400 μL of precipitant (methanol: acetonitrile = 2:1, V/V) to the SP:SAMPLEPREP_SUMMARY sample, seal the membrane with a film sealer, and shake for 1 min; 3 Place the SP:SAMPLEPREP_SUMMARY sample in ice water for ultrasonication for 10 min; 4 Place the sonicated sample SP:SAMPLEPREP_SUMMARY at -20°C for 2h; 5 Centrifuge the sample in a centrifuge (4,000 rpm * 4°C, 30 SP:SAMPLEPREP_SUMMARY min), each sample taken 300 μL to the new 96-well plate; 6 Put the 96-well SP:SAMPLEPREP_SUMMARY plate into a freeze dryer and drain it; 7 On the drained plate, add 300 μL of SP:SAMPLEPREP_SUMMARY 10% methanol, mix with a Multi-Tube vorterer (parameter setting 2,500HZ, 3 min), SP:SAMPLEPREP_SUMMARY and place in ice water for 10 min; 8 Centrifuge for 20 min (4,000 rpm * 4°C), SP:SAMPLEPREP_SUMMARY take 250 μL of the supernatant on a new plate; 9 For each sample (excluding SP:SAMPLEPREP_SUMMARY preparation QC), take 50 μL of each sample and mix into a QC sample, and the SP:SAMPLEPREP_SUMMARY prepared supernatant is ready to be used. SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters Acquity BEH HSS T3 (100 x 2.1mm, 1.8um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Waters Xevo-TQ MS:INSTRUMENT_TYPE QTOF MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS capillary and sampling cone voltages were set at 3.0 kV and 40.0 V, respectively MS:MS_RESULTS_FILE ST001858_AN003012_Results.txt UNITS:Peak Area Has m/z:Yes Has RT:Yes RT units:Minutes #END