#METABOLOMICS WORKBENCH silviaradenkovic_20230415_081530 DATATRACK_ID:3865 STUDY_ID:ST002563 ANALYSIS_ID:AN004224 PROJECT_ID:PR001653 VERSION 1 CREATED_ON April 18, 2023, 10:56 am #PROJECT PR:PROJECT_TITLE Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1 PR:PROJECT_SUMMARY Abnormal polyol metabolism has been predominantly associated with diabetes, PR:PROJECT_SUMMARY where excess glucose is converted to sorbitol by aldose reductase (AR). PR:PROJECT_SUMMARY Recently, abnormal polyol metabolism has also been implicated in PR:PROJECT_SUMMARY phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and PR:PROJECT_SUMMARY epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. PR:PROJECT_SUMMARY Given that the PMM enzyme is not closely connected to polyol metabolism, and, PR:PROJECT_SUMMARY unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the PR:PROJECT_SUMMARY increased polyol production, and the therapeutic mechanism of epalrestat in PR:PROJECT_SUMMARY PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM PR:PROJECT_SUMMARY enzyme and results in a depletion of mannose-1-P and guanosine diphosphate PR:PROJECT_SUMMARY mannose (GDP-mannose), which is essential for glycosylation. Here, we show that PR:PROJECT_SUMMARY apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in PR:PROJECT_SUMMARY intracellular glucose flux, which results in an increase in intracellular PR:PROJECT_SUMMARY polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts PR:PROJECT_SUMMARY glucose flux away from polyol production towards the synthesis of sugar PR:PROJECT_SUMMARY nucleotides. PR:INSTITUTE Mayo Clinic PR:LAST_NAME Radenkovic PR:FIRST_NAME Silvia PR:ADDRESS 200 2nd Ave SW Rochester MN PR:EMAIL radenkovic.silvia@mayo.edu PR:PHONE 507(77) 6-6107 PR:FUNDING_SOURCE NIH, KU Leuven #STUDY ST:STUDY_TITLE Metabolomic profiling of PMM2-CDG after siRNA mediated KD of AKR1b1 ST:STUDY_SUMMARY Abnormal polyol metabolism has been predominantly associated with diabetes, ST:STUDY_SUMMARY where excess glucose is converted to sorbitol by aldose reductase (AR). ST:STUDY_SUMMARY Recently, abnormal polyol metabolism has also been implicated in ST:STUDY_SUMMARY phosphomannomutase 2-congenital disorder of glycosylation (PMM2-CDG), and ST:STUDY_SUMMARY epalrestat, an AR inhibitor, proposed as a potential therapy for this disorder. ST:STUDY_SUMMARY Given that the PMM enzyme is not closely connected to polyol metabolism, and, ST:STUDY_SUMMARY unlike in diabetes, PMM2-CDG does not present with hyperglycemia in blood, the ST:STUDY_SUMMARY increased polyol production, and the therapeutic mechanism of epalrestat in ST:STUDY_SUMMARY PMM2-CDG remained largely elusive. PMM2-CDG is caused by deficiency of the PMM ST:STUDY_SUMMARY enzyme and results in a depletion of mannose-1-P and guanosine diphosphate ST:STUDY_SUMMARY mannose (GDP-mannose), which is essential for glycosylation. Here, we show that ST:STUDY_SUMMARY apart from glycosylation abnormalities, PMM2 deficiency also leads to changes in ST:STUDY_SUMMARY intracellular glucose flux, which results in an increase in intracellular ST:STUDY_SUMMARY polyols. Ssing tracer glucose studies, we demonstrate that AR inhibition diverts ST:STUDY_SUMMARY glucose flux away from polyol production towards the synthesis of sugar ST:STUDY_SUMMARY nucleotides. ST:INSTITUTE Mayo Clinic ST:LAST_NAME Radenkovic ST:FIRST_NAME Silvia ST:ADDRESS 200 2nd Ave SW Rochester MN, USA ST:EMAIL radenkovic.silvia@mayo.edu ST:PHONE 507(77) 6-6107 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENOTYPE_STRAIN WT/PMM2-CDG SU:AGE_OR_AGE_RANGE 5-45 SU:GENDER Male and female #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 P5 MCF0000858_SR01 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR01 SUBJECT_SAMPLE_FACTORS P5 MCF0000858_SR02 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR02 SUBJECT_SAMPLE_FACTORS P5 MCF0000858_SR03 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR03 SUBJECT_SAMPLE_FACTORS P5 MCF0000858_SR04 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR04 SUBJECT_SAMPLE_FACTORS P5 MCF0000858_SR05 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR05 SUBJECT_SAMPLE_FACTORS P5 MCF0000858_SR06 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR06 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR07 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR07 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR08 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR08 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR09 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR09 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR10 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR10 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR11 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR11 SUBJECT_SAMPLE_FACTORS C3 MCF0000858_SR12 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR12 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR13 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR13 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR14 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR14 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR15 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR15 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR16 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR16 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR17 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR17 SUBJECT_SAMPLE_FACTORS P2 MCF0000858_SR18 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR18 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR19 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR19 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR20 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR20 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR21 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR21 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR22 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR22 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR23 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR23 SUBJECT_SAMPLE_FACTORS C2 MCF0000858_SR24 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF0000858_SR24 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR01 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR01 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR02 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR02 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR03 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR03 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR04 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR04 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR05 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR05 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR06 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR06 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR07 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR07 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR08 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR08 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR09 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR09 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR10 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR10 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR11 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR11 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR12 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR12 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR13 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR13 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR14 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR14 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR15 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR15 SUBJECT_SAMPLE_FACTORS C3 MCF000867_SR16 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR16 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR17 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR17 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR18 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR18 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR19 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR19 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR20 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR20 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR21 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR21 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR22 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR22 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR23 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR23 SUBJECT_SAMPLE_FACTORS P5 MCF000867_SR24 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000867_SR24 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR01 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR01 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR02 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR02 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR03 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR03 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR04 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR04 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR05 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR05 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR06 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR06 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR07 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR07 SUBJECT_SAMPLE_FACTORS P2 MCF000887_SR08 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR08 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR09 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR09 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR10 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR10 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR11 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR11 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR12 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR12 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR13 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR13 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR14 Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR14 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR15 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR15 SUBJECT_SAMPLE_FACTORS P5 MCF000887_SR16 Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR16 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR17 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR17 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR18 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR18 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR19 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR19 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR20 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR20 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR21 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR21 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR22 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR22 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR23 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR23 SUBJECT_SAMPLE_FACTORS C3 MCF000887_SR24 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR24 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR25 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR25 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR26 Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR26 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR27 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR27 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR28 Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR28 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR29 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR29 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR30 Genotype:WT | Treatment:5.5mM 13C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR30 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR31 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR31 SUBJECT_SAMPLE_FACTORS C1 MCF000887_SR32 Genotype:WT | Treatment:5.5mM 12C Glc siRNA Tissue type=Fibroblasts; Organism=Homo sapiens; RAW_FILE_NAME=MCF000887_SR32 #COLLECTION CO:COLLECTION_SUMMARY Cells were washed with PBS, cells were incubated with extraction buffer for 2min CO:COLLECTION_SUMMARY before scraping and transferring to a fresh eppendorf. Samples were precipitated CO:COLLECTION_SUMMARY overnight at -80, then they were centrifuged at max rpm, 20min, 4 degrees C and CO:COLLECTION_SUMMARY supernatant transferred to an M/S vial. CO:SAMPLE_TYPE Fibroblasts CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Cells were treated with vehicle or 5nM siRNA targeting AKR1B1 #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites were extracted with 80% Methanol and IS (d27 myristic acid) SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY C18 iP REVERSE PHASE CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.8um) CH:SOLVENT_A 100% water; 10mM tributylamine; 15mM acetic acid CH:SOLVENT_B 100% methanol CH:FLOW_GRADIENT The gradient started with 5% of solvent B and 95% solvent A and remained at 5% B CH:FLOW_GRADIENT until 2 min post injection. A linear gradient to 37% B was carried out until 7 CH:FLOW_GRADIENT min and increased to 41% until 14 min. Between 14 and 26 minutes the gradient CH:FLOW_GRADIENT increased to 95% of B and remained at 95% B for 4 minutes. At 30 min the CH:FLOW_GRADIENT gradient returned to 5% B. The chromatography was stopped at 40 min. CH:FLOW_RATE 0.25 mL/min CH:COLUMN_TEMPERATURE 40 #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS El-Maven polly, ThermoFisher Xcalibur; Metabolites were annotated using the MS:MS_COMMENTS inhouse standard metabolite library- elution time and m/z values. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AUC MS_METABOLITE_DATA_START Samples MCF0000858_SR01 MCF0000858_SR02 MCF0000858_SR03 MCF0000858_SR04 MCF0000858_SR05 MCF0000858_SR06 MCF0000858_SR07 MCF0000858_SR08 MCF0000858_SR09 MCF0000858_SR10 MCF0000858_SR11 MCF0000858_SR12 MCF0000858_SR13 MCF0000858_SR14 MCF0000858_SR15 MCF0000858_SR16 MCF0000858_SR17 MCF0000858_SR18 MCF0000858_SR19 MCF0000858_SR20 MCF0000858_SR21 MCF0000858_SR22 MCF0000858_SR23 MCF0000858_SR24 MCF000867_SR01 MCF000867_SR02 MCF000867_SR03 MCF000867_SR04 MCF000867_SR05 MCF000867_SR06 MCF000867_SR07 MCF000867_SR08 MCF000867_SR09 MCF000867_SR10 MCF000867_SR11 MCF000867_SR12 MCF000867_SR13 MCF000867_SR14 MCF000867_SR15 MCF000867_SR16 MCF000867_SR17 MCF000867_SR18 MCF000867_SR19 MCF000867_SR20 MCF000867_SR21 MCF000867_SR22 MCF000867_SR23 MCF000867_SR24 MCF000887_SR01 MCF000887_SR02 MCF000887_SR03 MCF000887_SR04 MCF000887_SR05 MCF000887_SR06 MCF000887_SR07 MCF000887_SR08 MCF000887_SR09 MCF000887_SR10 MCF000887_SR11 MCF000887_SR12 MCF000887_SR13 MCF000887_SR14 MCF000887_SR15 MCF000887_SR16 MCF000887_SR17 MCF000887_SR18 MCF000887_SR19 MCF000887_SR20 MCF000887_SR21 MCF000887_SR22 MCF000887_SR23 MCF000887_SR24 MCF000887_SR25 MCF000887_SR26 MCF000887_SR27 MCF000887_SR28 MCF000887_SR29 MCF000887_SR30 MCF000887_SR31 MCF000887_SR32 Factors Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc negative siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 13C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:PMM2-CDG | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 12C Glc negative siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 13C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Genotype:WT | Treatment:5.5mM 12C Glc siRNA Sorbitol 6.20E+06 6.61E+06 1.52E+06 1.24E+06 1.87E+06 1.69E+06 4.35E+06 5.34E+06 1.12E+06 1.06E+06 1.42E+06 1.59E+06 5.38E+06 6.43E+06 1.47E+06 2.12E+06 1.79E+06 1.47E+06 2.65E+06 3.03E+06 2.05E+06 1.30E+06 1.96E+06 1.84E+06 7.30E+06 6.26E+06 7.74E+06 7.36E+06 1.88E+06 1.69E+06 2.00E+06 2.83E+06 5.72E+06 6.04E+06 7.08E+06 6.24E+06 2.28E+06 6.27E+06 2.16E+06 7.49E+06 6.98E+06 6.60E+06 7.21E+06 6.73E+06 3.25E+06 2.37E+06 2.78E+06 3.27E+06 1.12E+07 1.01E+07 1.12E+07 1.07E+07 2.12E+06 2.52E+06 2.51E+06 8.03E+06 9.22E+06 9.43E+06 1.28E+07 1.12E+07 2.12E+06 2.87E+06 2.22E+06 2.86E+06 1.02E+07 8.17E+06 9.08E+06 8.56E+06 1.91E+06 1.85E+06 4.25E+06 4.13E+06 8.11E+06 9.52E+06 1.11E+07 1.13E+07 1.97E+06 2.30E+06 2.56E+06 3.05E+06 CMP-sialic acid 4.47E+06 6.79E+06 7.11E+06 8.04E+06 1.08E+07 1.12E+07 8.28E+06 1.25E+07 1.25E+07 1.16E+07 1.62E+07 1.77E+07 5.86E+06 7.43E+06 5.86E+06 5.61E+06 9.42E+06 9.26E+06 7.15E+06 1.00E+07 6.83E+06 7.86E+06 1.09E+07 1.11E+07 4619724.28 5544198.58 8414899.53 11529223.49 3787443 3622040.96 7120839.87 7810364.68 8475719.63 8731786.6 13723911.76 13594607.82 13875730.68 12357849.2 17104895.19 20102595.26 5439974.33 5608650.69 9197733.82 6703395.13 10216174.44 11960217.6 19507225.28 18098294.68 2.19E+06 1.48E+06 4.86E+06 4.66E+06 2.60E+06 2.22E+06 5.97E+06 4.89E+06 3.09E+06 6.54E+05 3.24E+06 3.64E+06 1.96E+06 1.54E+06 4.88E+06 6.03E+06 4.85E+06 3.99E+06 1.02E+07 1.15E+07 7.70E+06 6.65E+06 1.33E+07 1.61E+07 1.84E+06 2.87E+06 2.79E+06 4.04E+06 9.64E+05 3.85E+06 3.87E+06 4.28E+06 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name parent formula M/Z HMDB ID isotopeLabel isotope count Sorbitol C6H14O6 181.07164 HMDB00247 C12 PARENT m0-6 CMP-sialic acid C20H31N4O16P 613.140442 HMDB0000230 C12 PARENT m0-20 METABOLITES_END #END