#METABOLOMICS WORKBENCH silviaradenkovic_20230414_133420 DATATRACK_ID:3863 STUDY_ID:ST002561 ANALYSIS_ID:AN004222 PROJECT_ID:PR001653
VERSION             	1
CREATED_ON             	April 18, 2023, 10:44 am
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic profiling of PMM2-CDG zebrafish in presence and absence of
PR:PROJECT_TITLE                 	epalrestat
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. Targeting AR with epalrestat decreases polyol levels and increases
PR:PROJECT_SUMMARY               	GDP-mannose in vivo in pmm2 mutant zebrafish.
PR:INSTITUTE                     	Mayo Clinic
PR:LAST_NAME                     	Radenkovic
PR:FIRST_NAME                    	Silvia
PR:ADDRESS                       	200 2nd Ave SW Rochester MN, USA
PR:EMAIL                         	radenkovic.silvia@mayo.edu
PR:PHONE                         	507(77) 6-6107
PR:FUNDING_SOURCE                	NIH, KU Leuven
PR:PUBLICATIONS                  	Tracer metabolomics reveals the role of aldose reductase in glycosylation
PR:CONTRIBUTORS                  	Silvia Radenkovic, Anna N. Ligezka, Sneha S. Mokashi, Karen Driesen, Lynn
PR:CONTRIBUTORS                  	Dukes-Rimsky, Graeme Preston, Luckio F. Owuocha, Leila Sabbagh, Jehan Mousa,
PR:CONTRIBUTORS                  	Christina Lam, Andrew Edmondson, Austin Larson, Matthew Schultz, Pieter
PR:CONTRIBUTORS                  	Vermeersch, David Cassiman, Peter Witters, Lesa J. Beamer, Tamas Kozicz, Heather
PR:CONTRIBUTORS                  	Flanagan-Steet, Bart Ghesquière, Eva Morava
#STUDY
ST:STUDY_TITLE                   	Metabolomic profiling of PMM2-CDG zebrafish in presence and absence of
ST:STUDY_TITLE                   	epalrestat
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. Targeting AR with epalrestat decreases polyol levels and increases
ST:STUDY_SUMMARY                 	GDP-mannose in vivo in pmm2 mutant zebrafish.
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                  	Fish
SU:SUBJECT_SPECIES               	Danio rerio
SU:TAXONOMY_ID                   	7955
SU:GENOTYPE_STRAIN               	WT/PMM2 mutant sa10150
SU:AGE_OR_AGE_RANGE              	6 dpf
#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           	-	MCF001027_wt1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt1
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt2
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt3
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt4
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_wt3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_wt3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm1
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm2
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm3
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm4
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001027_pmm4_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=MCF001027_pmm4_ep40
SUBJECT_SAMPLE_FACTORS           	-	wt1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt1
SUBJECT_SAMPLE_FACTORS           	-	wt2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt2
SUBJECT_SAMPLE_FACTORS           	-	wt3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt3
SUBJECT_SAMPLE_FACTORS           	-	wt1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt1_ep10
SUBJECT_SAMPLE_FACTORS           	-	wt2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt2_ep10
SUBJECT_SAMPLE_FACTORS           	-	wt3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt3_ep10
SUBJECT_SAMPLE_FACTORS           	-	wt4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt4_ep10
SUBJECT_SAMPLE_FACTORS           	-	pmm1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm1
SUBJECT_SAMPLE_FACTORS           	-	pmm2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm2
SUBJECT_SAMPLE_FACTORS           	-	pmm3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm3
SUBJECT_SAMPLE_FACTORS           	-	pmm4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm4
SUBJECT_SAMPLE_FACTORS           	-	pmm1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm1_ep10
SUBJECT_SAMPLE_FACTORS           	-	pmm2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm2_ep10
SUBJECT_SAMPLE_FACTORS           	-	pmm3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm3_ep10
SUBJECT_SAMPLE_FACTORS           	-	pmm4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm4_ep10
SUBJECT_SAMPLE_FACTORS           	-	wt4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt4
SUBJECT_SAMPLE_FACTORS           	-	wt5	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt5
SUBJECT_SAMPLE_FACTORS           	-	wt1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt1_ep40
SUBJECT_SAMPLE_FACTORS           	-	wt2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt2_ep40
SUBJECT_SAMPLE_FACTORS           	-	wt3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=HiliC; RAW_FILE_NAME=wt3_ep40
SUBJECT_SAMPLE_FACTORS           	-	pmm5	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm5
SUBJECT_SAMPLE_FACTORS           	-	pmm6	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm6
SUBJECT_SAMPLE_FACTORS           	-	pmm7	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm7
SUBJECT_SAMPLE_FACTORS           	-	pmm8	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm8
SUBJECT_SAMPLE_FACTORS           	-	pmm1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm1_ep40
SUBJECT_SAMPLE_FACTORS           	-	pmm2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm2_ep40
SUBJECT_SAMPLE_FACTORS           	-	pmm3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm3_ep40
SUBJECT_SAMPLE_FACTORS           	-	pmm4_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=HiliC; RAW_FILE_NAME=pmm4_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt1
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt2
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt4_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt4_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm1
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm2
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt3
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt4
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt3_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_wt4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_wt4_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm3
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm4	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm4
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm3_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm4_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001395_pmm4_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001395_pmm4_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt1
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt2
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt3
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt3_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt4_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_wt3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio wildtype; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_wt3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm1	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm1
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm2	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm2
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm3	Sample treatment:DMSO	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm3
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm1_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm1_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm2_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm2_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm3_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm3_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm4_ep10	Sample treatment:10µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm4_ep10
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm1_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm1_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm2_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm2_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm3_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm3_ep40
SUBJECT_SAMPLE_FACTORS           	-	MCF001428_pmm4_ep40	Sample treatment:40µM Epalrestat	Organism part=whole organism; Organism type=zebrafish; Organism=Danio rerio PMM2 sa10150 mutant; Chromatography=Reverse phase IP; RAW_FILE_NAME=MCF001428_pmm4_ep40
#COLLECTION
CO:COLLECTION_SUMMARY            	10 embryos per sample were homogenized by sonication in extraction buffer (80%
CO:COLLECTION_SUMMARY            	MeOH, IS). The metabolites were then precipitated at -80 °C overnight, after
CO:COLLECTION_SUMMARY            	which the samples were centrifuged at 15,000 rpm, 20 min, 4 °C. Next, the
CO:COLLECTION_SUMMARY            	supernatant was transferred to a fresh Eppendorf tube. The samples were
CO:COLLECTION_SUMMARY            	centrifuged again at 15,000 rpm, 20 min, 4 °C to remove any leftover impurities
CO:COLLECTION_SUMMARY            	and transferred to a fresh MS vial. Finally, the samples were analyzed by LC/MS
CO:SAMPLE_TYPE                   	Larvae
CO:STORAGE_CONDITIONS            	-80?
#TREATMENT
TR:TREATMENT_SUMMARY             	Zebrafish were treated with vehicle or 10 or 40 microM epalrestat
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	10 embryos per sample were homogenized by sonication in extraction buffer (80%
SP:SAMPLEPREP_SUMMARY            	MeOH, IS). The metabolites were then precipitated at -80 °C overnight, after
SP:SAMPLEPREP_SUMMARY            	which the samples were centrifuged at 15,000 rpm, 20 min, 4 °C. Next, the
SP:SAMPLEPREP_SUMMARY            	supernatant was transferred to a fresh Eppendorf tube. The samples were
SP:SAMPLEPREP_SUMMARY            	centrifuged again at 15,000 rpm, 20 min, 4 °C to remove any leftover impurities
SP:SAMPLEPREP_SUMMARY            	and transferred to a fresh MS vial. Finally, the samples were analyzed by LC/MS
SP:PROCESSING_STORAGE_CONDITIONS 	-80?
SP:EXTRACTION_METHOD             	80 % MeOH with Internal standards (e.g. d27 myristic acid)
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	HILIC
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Agilent Infinity
CH:COLUMN_NAME                   	Poroshell 120 HILIC-Z PEEK Column 2.1x 150mm, 5 um
CH:SOLVENT_A                     	100% acetonitrile
CH:SOLVENT_B                     	100% water; 10 mM sodium acetate, pH 9.3
CH:FLOW_GRADIENT                 	A linear gradient was carried out starting with 90% solvent A and 10% solvent B.
CH:FLOW_GRADIENT                 	From 2 to 12 min the gradient changed to 60% B. The gradient was kept on 60% B
CH:FLOW_GRADIENT                 	for 3 minutes and followed by a decrease to 10% B. The chromatography was
CH:FLOW_GRADIENT                 	stopped at 25 min
CH:FLOW_RATE                     	0.25 mL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Focus
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	El-Maven polly, ThemoFisher Xcalibur Metabolites were annotated using in-house
MS:MS_COMMENTS                   	metabolite library- elution time and m/z were considered
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	AUC
MS_METABOLITE_DATA_START
Samples	MCF001027_wt1	MCF001027_wt2	MCF001027_wt3	MCF001027_wt4	MCF001027_wt1_ep10	MCF001027_wt2_ep10	MCF001027_wt1_ep40	MCF001027_wt2_ep40	MCF001027_wt3_ep40	MCF001027_pmm1	MCF001027_pmm2	MCF001027_pmm3	MCF001027_pmm4	MCF001027_pmm1_ep10	MCF001027_pmm2_ep10	MCF001027_pmm1_ep40	MCF001027_pmm2_ep40	MCF001027_pmm3_ep40	MCF001027_pmm4_ep40	wt1	wt2	wt3	wt1_ep10	wt2_ep10	wt3_ep10	wt4_ep10	pmm1	pmm2	pmm3	pmm4	pmm1_ep10	pmm2_ep10	pmm3_ep10	pmm4_ep10	wt4	wt5	wt1_ep40	wt2_ep40	wt3_ep40	pmm5	pmm6	pmm7	pmm8	pmm1_ep40	pmm2_ep40	pmm3_ep40	pmm4_ep40
Factors	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat
Sorbitol	1.26E+06	1.40E+06	1.17E+06	9.85E+05	9.05E+05	1.35E+06	8.93E+05	8.26E+05	1.04E+06	1.49E+06	1.43E+06	1.48E+06	1.41E+06	1.42E+06	1.61E+06	1.28E+06	1.03E+06	1.50E+06	7.78E+05	3.06E+06	3.79E+06	2.92E+06	3.07E+06	3.04E+06	3.17E+06	2.84E+06	3.46E+06	3.50E+06	4.48E+06	3.08E+06	3.16E+06	2.98E+06	3.36E+06	2.95E+06	2.82E+06	2.91E+06	2.03E+06	1.90E+06	2.00E+06	2.43E+06	3.31E+06	2.77E+06	2.66E+06	2.19E+06	1.97E+06	1.80E+06	1.96E+06
GDP-mannose	3.03E+05	1.81E+05	1.76E+05	2.27E+05	3.16E+05	3.11E+05	2.82E+05	3.01E+05	3.46E+05	5.35E+04	9.92E+04	7.64E+04	6.00E+04	1.39E+05	1.44E+05	1.21E+05	1.44E+05	1.90E+05	2.99E+05	8.37E+05	8.28E+05	6.91E+05	7.77E+05	7.37E+05	7.90E+05	7.16E+05	5.19E+05	5.29E+05	3.44E+05	3.38E+05	5.50E+05	5.07E+05	5.30E+05	3.66E+05	6.76E+05	5.95E+05	5.19E+05	6.18E+05	6.81E+05	4.80E+05	3.77E+05	4.50E+05	4.25E+05	4.23E+05	4.58E+05	3.08E+05	3.63E+05
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
GDP-mannose	C16H25N5O16P2	604.06897	HMDB01163	C12 PARENT	m0
METABOLITES_END
#END