#METABOLOMICS WORKBENCH silviaradenkovic_20230414_133420 DATATRACK_ID:3863 STUDY_ID:ST002561 ANALYSIS_ID:AN004221 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        	Reverse phase, IP, C18 column
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	C-18: Acquity UPLC -HSS T3 1. 8 µm; 2.1 x 150 mm, Waters
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, 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	MCF001395_wt1	MCF001395_wt2	MCF001395_wt1_ep10	MCF001395_wt2_ep10	MCF001395_wt1_ep40	MCF001395_wt2_ep40	MCF001395_wt3_ep40	MCF001395_wt4_ep40	MCF001395_pmm1	MCF001395_pmm2	MCF001395_pmm1_ep10	MCF001395_pmm2_ep10	MCF001395_pmm1_ep40	MCF001395_pmm2_ep40	MCF001395_wt3	MCF001395_wt4	MCF001395_wt3_ep10	MCF001395_wt4_ep10	MCF001395_pmm3	MCF001395_pmm4	MCF001395_pmm3_ep10	MCF001395_pmm4_ep10	MCF001395_pmm3_ep40	MCF001395_pmm4_ep40	MCF001428_wt1	MCF001428_wt2	MCF001428_wt3	MCF001428_wt1_ep10	MCF001428_wt2_ep10	MCF001428_wt3_ep10	MCF001428_wt4_ep10	MCF001428_wt1_ep40	MCF001428_wt2_ep40	MCF001428_wt3_ep40	MCF001428_pmm1	MCF001428_pmm2	MCF001428_pmm3	MCF001428_pmm1_ep10	MCF001428_pmm2_ep10	MCF001428_pmm3_ep10	MCF001428_pmm4_ep10	MCF001428_pmm1_ep40	MCF001428_pmm2_ep40	MCF001428_pmm3_ep40	MCF001428_pmm4_ep40
Factors	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:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM Epalrestat	Sample treatment:DMSO	Sample treatment:DMSO	Sample treatment:10µM Epalrestat	Sample treatment:10µM 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: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:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat	Sample treatment:40µM Epalrestat
Sorbitol	2.52E+06	2.45E+06	2.41E+06	2.73E+06	2.41E+06	2.16E+06	2.26E+06	2.23E+06	2.45E+06	2.52E+06	2.34E+06	1.98E+06	2.10E+06	1.61E+06	2.01E+06	1.87E+06	1.79E+06	1.98E+06	1.96E+06	1.92E+06	1.53E+06	1.62E+06	1.49E+06	1.57E+06	7.69E+06	1.05E+07	8.85E+06	1.03E+07	7.54E+06	7.02E+06	9.31E+06	8.31E+06	7.14E+06	7.64E+06	1.31E+07	1.96E+07	1.49E+07	1.45E+07	1.06E+07	1.60E+07	1.34E+07	1.36E+07	1.85E+07	1.36E+07	1.87E+07
GDP-mannose	2.93E+05	3.10E+05	2.96E+05	2.89E+05	2.99E+05	2.75E+05	3.28E+05	2.88E+05	2.04E+05	1.69E+05	1.90E+05	1.73E+05	1.50E+05	1.62E+05	3.13E+05	2.86E+05	2.24E+05	2.98E+05	1.76E+05	1.81E+05	1.97E+05	1.71E+05	1.64E+05	1.59E+05	1.30E+07	1.31E+07	1.42E+07	1.38E+07	1.32E+07	1.34E+07	1.31E+07	1.40E+07	1.21E+07	1.18E+07	8.06E+06	7.02E+06	1.00E+07	7.75E+06	1.00E+07	8.86E+06	1.09E+07	8.30E+06	8.51E+06	9.21E+06	8.35E+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
GDP-mannose	C16H25N5O16P2	604.06897	HMDB01163	C12 PARENT	m0
METABOLITES_END
#END