#METABOLOMICS WORKBENCH silviaradenkovic_20230421_122617 DATATRACK_ID:3880 STUDY_ID:ST002575 ANALYSIS_ID:AN004240 PROJECT_ID:PR001660 VERSION 1 CREATED_ON April 23, 2023, 2:45 pm #PROJECT PR:PROJECT_TITLE Metabolomic profiling of PMM2-CDG patient fibroblasts by GC/MS 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 such as sorbitol and mannitol. 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 patient fibroblasts by GC/MS 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 such as sorbitol and mannitol. 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 P1 Polyol_SR01 (1) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (1) SUBJECT_SAMPLE_FACTORS P1 Polyol_SR01 (2) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (2) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR01 (3) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (3) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR01 (4) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (4) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR01 (5) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (5) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR01 (6) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (6) SUBJECT_SAMPLE_FACTORS C2 Polyol_SR01 (7) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (7) SUBJECT_SAMPLE_FACTORS C2 Polyol_SR01 (8) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (8) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR01 (9) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (9) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR01 (10) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (10) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR01 (11) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (11) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR01 (12) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (12) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR01 (13) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (13) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR01 (14) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (14) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR01 (15) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (15) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR01 (16) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (16) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR01 (17) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (17) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR01 (18) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (18) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR01 (19) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (19) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR01 (20) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (20) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR01 (21) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (21) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR01 (22) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (22) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR01 (23) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (23) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR01 (24) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (24) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR01 (25) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (25) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR01 (26) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (26) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR01 (27) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (27) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR01 (28) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR01 (28) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (1) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (1) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (2) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (2) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (3) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (3) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (4) Genotype:PMM2-CDG | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (4) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (5) Genotype:PMM2-CDG | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (5) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (6) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (6) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (7) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (7) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (8) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (8) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (9) Genotype:WT | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (9) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (10) Genotype:WT | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (10) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (11) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (11) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (12) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (12) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (13) Genotype:PMM2-CDG | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (13) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (14) Genotype:PMM2-CDG | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (14) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (15) Genotype:PMM2-CDG | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (15) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (16) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (16) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (17) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (17) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (18) Genotype:WT | Treatment:Glucose+Fructose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (18) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (19) Genotype:WT | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (19) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (20) Genotype:WT | Treatment:Glucose Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (20) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (21) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (21) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (22) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (22) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (23) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (23) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (24) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (24) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (25) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (25) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (26) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (26) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (27) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (27) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (28) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (28) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (29) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (29) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR02 (30) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (30) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (31) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (31) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (32) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (32) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (33) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (33) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR02 (34) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR02 (34) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR03 (1) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (1) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR03 (2) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (2) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR03 (3) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (3) SUBJECT_SAMPLE_FACTORS P5 Polyol_SR03 (4) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (4) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR03 (5) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (5) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR03 (6) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (6) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR03 (7) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (7) SUBJECT_SAMPLE_FACTORS P2 Polyol_SR03 (8) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (8) SUBJECT_SAMPLE_FACTORS P1 Polyol_SR03 (9) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (9) SUBJECT_SAMPLE_FACTORS P1 Polyol_SR03 (10) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (10) SUBJECT_SAMPLE_FACTORS P1 Polyol_SR03 (11) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (11) SUBJECT_SAMPLE_FACTORS P1 Polyol_SR03 (12) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (12) SUBJECT_SAMPLE_FACTORS C4 Polyol_SR03 (13) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (13) SUBJECT_SAMPLE_FACTORS C4 Polyol_SR03 (14) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (14) SUBJECT_SAMPLE_FACTORS C4 Polyol_SR03 (15) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (15) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR03 (16) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (16) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR03 (17) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (17) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR03 (18) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (18) SUBJECT_SAMPLE_FACTORS C3 Polyol_SR03 (19) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR03 (19) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR04 (1) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (1) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR04 (2) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (2) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR04 (3) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (3) SUBJECT_SAMPLE_FACTORS P4 Polyol_SR04 (4) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (4) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR04 (5) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (5) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR04 (6) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (6) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR04 (7) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (7) SUBJECT_SAMPLE_FACTORS P3 Polyol_SR04 (8) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (8) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR04 (9) Genotype:PMM2-CDG | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (9) SUBJECT_SAMPLE_FACTORS P6 Polyol_SR04 (10) Genotype:PMM2-CDG | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (10) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR04 (11) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (11) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR04 (12) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (12) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR04 (13) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (13) SUBJECT_SAMPLE_FACTORS C1 Polyol_SR04 (14) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (14) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR04 (15) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (15) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR04 (16) Genotype:WT | Treatment:10microM epalrestat Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (16) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR04 (17) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (17) SUBJECT_SAMPLE_FACTORS C5 Polyol_SR04 (18) Genotype:WT | Treatment:Vehicle Cell type=Skin fibroblasts; RAW_FILE_NAME=Polyol_SR04 (18) #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 either 0 or 10microM epalrestat #SAMPLEPREP SP:SAMPLEPREP_SUMMARY metabolites were extracted from cells with 2 mL 80 % MeOH, IS and the SP:SAMPLEPREP_SUMMARY supernatant was transferred to a fresh 2 mL Eppendorf tube. Standards containing SP:SAMPLEPREP_SUMMARY metabolites of interest were prepared simultaneously and dried overnight SP:SAMPLEPREP_SUMMARY together with cellular extracts by vacuum centrifugation at 4 °C. The following SP:SAMPLEPREP_SUMMARY day, 20 µL of methoxyamine (MOX) (Sigma Aldrich) was added to the samples and SP:SAMPLEPREP_SUMMARY they were incubated for 90 min at 37 °C. Next, 60 µL of N, SP:SAMPLEPREP_SUMMARY O-bis(trimethylsilyl)trifluoroacetamide (TMS) (Sigma Aldrich) was added to the SP:SAMPLEPREP_SUMMARY samples, which were then incubated at 60 °C for 30 min. Samples were kept SP:SAMPLEPREP_SUMMARY overnight in a dry and cool place to allow further derivatization with TMS SP:PROCESSING_STORAGE_CONDITIONS -80℃ SP:EXTRACT_STORAGE -80℃ SP:SAMPLE_DERIVATIZATION MOX, TMS #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Agilent 7890A GC (Agilent Technologies) coupled with an HP-5 ms 5 % phenyl CH:CHROMATOGRAPHY_SUMMARY methyl silox capillary column (30 m, 0.25 mm, 0.25 um, Agilent Technologies) CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME Agilent HP-5ms (30m x 0.25mm, 0.25um) CH:SOLVENT_A N/A CH:SOLVENT_B N/A CH:FLOW_GRADIENT The temperature gradient applied was as follows: 2 min 100°C, next 175 °C at CH:FLOW_GRADIENT Δ20 °C/min, followed by an increase to 230 °C at Δ4 °C/min, kept at 230 °C CH:FLOW_GRADIENT for 3 min, increased to 300 °C at Δ 40°C/min and lastly kept at 300 °C for 5 CH:FLOW_GRADIENT min. The column was further baked for another 3 min at 325 °C after the CH:FLOW_GRADIENT gradient. CH:FLOW_RATE NA CH:COLUMN_TEMPERATURE Gradient #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Agilent 7000B MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE EI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Mass Hunter Workstation software with the Quantitative Analysis Version MS:MS_COMMENTS B.06.00/Build 6.0.388.0 Specific metabolites were identified based on their MS:MS_COMMENTS fragment formula (SIM)/elution time #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS AUC MS_METABOLITE_DATA_START Samples Polyol_SR01 (1) Polyol_SR01 (2) Polyol_SR01 (3) Polyol_SR01 (4) Polyol_SR01 (5) Polyol_SR01 (6) Polyol_SR01 (7) Polyol_SR01 (8) Polyol_SR01 (9) Polyol_SR01 (10) Polyol_SR01 (11) Polyol_SR01 (12) Polyol_SR01 (13) Polyol_SR01 (14) Polyol_SR01 (15) Polyol_SR01 (16) Polyol_SR01 (17) Polyol_SR01 (18) Polyol_SR01 (19) Polyol_SR01 (20) Polyol_SR01 (21) Polyol_SR01 (22) Polyol_SR01 (23) Polyol_SR01 (24) Polyol_SR01 (25) Polyol_SR01 (26) Polyol_SR01 (27) Polyol_SR01 (28) Polyol_SR02 (1) Polyol_SR02 (2) Polyol_SR02 (3) Polyol_SR02 (4) Polyol_SR02 (5) Polyol_SR02 (6) Polyol_SR02 (7) Polyol_SR02 (8) Polyol_SR02 (9) Polyol_SR02 (10) Polyol_SR02 (11) Polyol_SR02 (12) Polyol_SR02 (13) Polyol_SR02 (14) Polyol_SR02 (15) Polyol_SR02 (16) Polyol_SR02 (17) Polyol_SR02 (18) Polyol_SR02 (19) Polyol_SR02 (20) Polyol_SR02 (21) Polyol_SR02 (22) Polyol_SR02 (23) Polyol_SR02 (24) Polyol_SR02 (25) Polyol_SR02 (26) Polyol_SR02 (27) Polyol_SR02 (28) Polyol_SR02 (29) Polyol_SR02 (30) Polyol_SR02 (31) Polyol_SR02 (32) Polyol_SR02 (33) Polyol_SR02 (34) Polyol_SR03 (1) Polyol_SR03 (2) Polyol_SR03 (3) Polyol_SR03 (4) Polyol_SR03 (5) Polyol_SR03 (6) Polyol_SR03 (7) Polyol_SR03 (8) Polyol_SR03 (9) Polyol_SR03 (10) Polyol_SR03 (11) Polyol_SR03 (12) Polyol_SR03 (13) Polyol_SR03 (14) Polyol_SR03 (15) Polyol_SR03 (16) Polyol_SR03 (17) Polyol_SR03 (18) Polyol_SR03 (19) Polyol_SR04 (1) Polyol_SR04 (2) Polyol_SR04 (3) Polyol_SR04 (4) Polyol_SR04 (5) Polyol_SR04 (6) Polyol_SR04 (7) Polyol_SR04 (8) Polyol_SR04 (9) Polyol_SR04 (10) Polyol_SR04 (11) Polyol_SR04 (12) Polyol_SR04 (13) Polyol_SR04 (14) Polyol_SR04 (15) Polyol_SR04 (16) Polyol_SR04 (17) Polyol_SR04 (18) Factors Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose Genotype:PMM2-CDG | Treatment:Glucose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose Genotype:WT | Treatment:Glucose Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose+Fructose Genotype:PMM2-CDG | Treatment:Glucose Genotype:PMM2-CDG | Treatment:Glucose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose+Fructose Genotype:WT | Treatment:Glucose Genotype:WT | Treatment:Glucose Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:Vehicle Genotype:PMM2-CDG | Treatment:10microM epalrestat Genotype:PMM2-CDG | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:10microM epalrestat Genotype:WT | Treatment:Vehicle Genotype:WT | Treatment:Vehicle Sorbitol 93349.76178 466496.2863 57880.45374 80840.54602 96132.85896 102686.5611 70925.95768 95782.86832 84560.63631 134609.2893 91063.95489 91153.66594 104269.926 112738.9167 155947.4556 174983.0563 71793.69938 113116.8802 80535.95306 70425.76902 183369.2035 246629.3608 190946.6136 168158.0474 159524.6488 217042.5494 410204.4591 150078.9837 251028.018 36640.73689 22729.67882 106478.4358 21203.23642 56904.7806 36964.41901 42794.97079 32328.13347 37253.44069 2228029.899 1788172.074 1353796.64 1918388.101 1412972.721 1815576.62 1064561.568 1156773.356 1376227.714 1343770.664 17460.74132 106478.4358 21203.23642 17460.74132 270306.0499 106478.4358 21203.23642 24912.95432 129800.462 28398.34021 27687.77846 54039.69285 57823.71013 51669.22057 3.66E+06 3.34E+06 5.34E+06 4.16E+06 3.38E+06 3.28E+06 4.75E+06 3.76E+06 1.83E+06 1.67E+06 2.44E+06 2.43E+06 2.64E+06 2.08E+06 2.14E+06 2.05E+06 2.14E+06 2.54E+06 2.41E+06 29331.2563 9000.00887 15380.41772 18772.67088 12013.77572 12351.78298 62593.66745 11039.56093 22264.73781 16527.89409 13345.2292 9871.729501 12265.3907 13103.81222 7866.283744 8179.43839 9142.855318 11417.05308 Mannitol 5532.40909 36119.01096 2171.058651 3927.551931 3149.222605 2594.084347 2101.761759 3249.961498 2030.265207 2766.065386 3011.870016 6289.841792 3518.790547 4311.811028 3371.570937 3676.23737 2643.602087 10858.4253 3133.06996 3530.917937 5189.191233 14968.10001 2262.452534 4295.372263 2115.377574 10247.91913 22627.67277 2762.49567 1965871.244 1680647.896 993322.184 1522674.29 1328962.383 1534577.644 910492.1983 988665.705 949302.5863 1058534.903 302839.4924 48014.50429 36715.21909 129800.462 28398.34021 71748.2823 39578.62717 52039.79851 57823.71013 51669.22057 993568.6869 1522674.29 1328962.383 993568.6869 1344670.65 1522674.29 1328962.383 1378047.78 1918388.101 1412972.721 1009834.113 1293286.636 1376227.714 1343770.664 87361.64056 132059.4298 171499.2396 104210.4847 93153.71556 100285.6905 92156.95062 93667.66803 68760.63274 95030.42728 90916.3403 88452.54893 75350.46249 94026.18613 57199.41826 67745.34501 87474.89292 67605.95955 91588.92809 571222.7415 519906.8595 789698.4226 750750.7501 877422.3837 725558.5589 936745.8792 810468.8197 328095.3975 320085.9055 409146.8169 316101.1657 528657.3745 469866.9203 222875.9792 209801.5534 252577.4917 272835.9025 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name parent formula SIM HMDB ID isotopeLabel isotope count RT Sorbitol C6H14O6 319 HMDB00247 C12 PARENT_6TMS m0-4 13.092 Mannitol C6H14O6 319 HMDB0000765 C12 PARENT_TMS m0-m4 12.959 METABOLITES_END #END