#METABOLOMICS WORKBENCH dj2122_20240830_065411 DATATRACK_ID:5157 STUDY_ID:ST003507 ANALYSIS_ID:AN005758 PROJECT_ID:PR002153
VERSION             	1
CREATED_ON             	September 8, 2024, 1:18 am
#PROJECT
PR:PROJECT_TITLE                 	A NRF2/β3-adrenoreceptor axis drives a sustained antioxidant and metabolic
PR:PROJECT_TITLE                 	rewiring through the pentose-phosphate pathway to alleviate cardiac stress
PR:PROJECT_SUMMARY               	Background Cardiac β3-adrenergic receptors (β3AR) are upregulated in diseased
PR:PROJECT_SUMMARY               	hearts and mediate antithetic effects to those of β1AR and β2AR. β3AR
PR:PROJECT_SUMMARY               	agonists were recently shown to protect from myocardial remodeling in
PR:PROJECT_SUMMARY               	preclinical studies and to improve systolic function in patients with severe
PR:PROJECT_SUMMARY               	heart failure. The underlying mechanisms, however, remain elusive. Methods To
PR:PROJECT_SUMMARY               	dissect functional, transcriptional and metabolic effects, hearts and isolated
PR:PROJECT_SUMMARY               	ventricular myocytes from mice harboring a moderate, cardiac-specific expression
PR:PROJECT_SUMMARY               	of a human ADRB3 transgene (β3AR-Tg) and subjected to transverse aortic
PR:PROJECT_SUMMARY               	constriction (TAC) were assessed using echocardiography, RNAseq, PET scan,
PR:PROJECT_SUMMARY               	metabolomics, seahorse and metabolic flux analysis. Subsequently, signaling and
PR:PROJECT_SUMMARY               	metabolic pathways were investigated further in vivo in β3AR-Tg and in vitro in
PR:PROJECT_SUMMARY               	neonatal rat ventricular myocytes adenovirally infected to express β3AR and
PR:PROJECT_SUMMARY               	subjected to neurohormonal stress. These results were completed with an analysis
PR:PROJECT_SUMMARY               	of single nucleus RNAseq data from human cardiac myocytes from heart failure
PR:PROJECT_SUMMARY               	patients. Results Compared with WT littermate, β3AR-Tg mice were protected from
PR:PROJECT_SUMMARY               	hypertrophy after transaortic constriction (TAC), while systolic function was
PR:PROJECT_SUMMARY               	preserved. β3AR-expressing hearts displayed enhanced myocardial glucose uptake
PR:PROJECT_SUMMARY               	under stress in absence of increased lactate levels. Instead, metabolomic and
PR:PROJECT_SUMMARY               	metabolic flux analyses in stressed hearts revealed an increase in intermediates
PR:PROJECT_SUMMARY               	of the Pentose-Phosphate Pathway (PPP) in β3AR-Tg, an alternative route of
PR:PROJECT_SUMMARY               	glucose utilization, paralleled with increased transcript levels of
PR:PROJECT_SUMMARY               	NADPH-producing and rate-limiting enzymes of the PPP, without fueling the
PR:PROJECT_SUMMARY               	hexosamine metabolism. The ensuing increased content of NADPH and of reduced
PR:PROJECT_SUMMARY               	glutathione decreased myocyte oxidant stress, while downstream oxidative
PR:PROJECT_SUMMARY               	metabolism assessed by oxygen consumption was preserved with higher glucose
PR:PROJECT_SUMMARY               	oxidation in β3AR-Tg post-TAC compared to WT, together with increased
PR:PROJECT_SUMMARY               	mitochondrial biogenesis. Unbiased transcriptomics and pathway analysis
PR:PROJECT_SUMMARY               	identified NRF2 (NFE2L2) as upstream transcription factor which was functionally
PR:PROJECT_SUMMARY               	verified in β3AR-expressing cardiac myocytes where its translocation and
PR:PROJECT_SUMMARY               	nuclear activity was dependent on β3AR activation of nitric-oxide synthase
PR:PROJECT_SUMMARY               	(NOS) NO production. Conclusion Moderate expression of cardiac β3AR, at levels
PR:PROJECT_SUMMARY               	observed in human cardiac myocardium, exerts antioxidant effects through
PR:PROJECT_SUMMARY               	activation of the PPP and NRF2 pathway, thereby preserving myocardial oxidative
PR:PROJECT_SUMMARY               	metabolism, function and integrity under pathophysiological stress.
PR:INSTITUTE                     	UCLouvain
PR:LAST_NAME                     	Dewulf
PR:FIRST_NAME                    	Joseph
PR:ADDRESS                       	Avenue Hippocrate 10, Brussels, Brussels, 1200, Belgium
PR:EMAIL                         	joseph.dewulf@uclouvain.be
PR:PHONE                         	027646727
#STUDY
ST:STUDY_TITLE                   	A NRF2/β3-adrenoreceptor axis drives a sustained antioxidant and metabolic
ST:STUDY_TITLE                   	rewiring through the pentose-phosphate pathway to alleviate cardiac stress
ST:STUDY_SUMMARY                 	WT TAC vs TG TAC vs WT baseline vs TG baseline Background: Cardiac
ST:STUDY_SUMMARY                 	β3-adrenergic receptors (β3AR) are upregulated in diseased hearts and mediate
ST:STUDY_SUMMARY                 	antithetic effects to those of β1AR and β2AR. β3AR agonists were recently
ST:STUDY_SUMMARY                 	shown to protect from myocardial remodeling in preclinical studies and to
ST:STUDY_SUMMARY                 	improve systolic function in patients with severe heart failure. The underlying
ST:STUDY_SUMMARY                 	mechanisms, however, remain elusive. Methods: To dissect functional,
ST:STUDY_SUMMARY                 	transcriptional and metabolic effects, hearts and isolated ventricular myocytes
ST:STUDY_SUMMARY                 	from mice harboring a moderate, cardiac-specific expression of a human ADRB3
ST:STUDY_SUMMARY                 	transgene (β3AR-Tg) and subjected to transverse aortic constriction (TAC) were
ST:STUDY_SUMMARY                 	assessed using echocardiography, RNAseq, PET scan, metabolomics, seahorse and
ST:STUDY_SUMMARY                 	metabolic flux analysis. Subsequently, signaling and metabolic pathways were
ST:STUDY_SUMMARY                 	investigated further in vivo in β3AR-Tg and in vitro in neonatal rat
ST:STUDY_SUMMARY                 	ventricular myocytes adenovirally infected to express β3AR and subjected to
ST:STUDY_SUMMARY                 	neurohormonal stress. These results were completed with an analysis of single
ST:STUDY_SUMMARY                 	nucleus RNAseq data from human cardiac myocytes from heart failure patients.
ST:STUDY_SUMMARY                 	Results: Compared with WT littermate, β3AR-Tg mice were protected from
ST:STUDY_SUMMARY                 	hypertrophy after transaortic constriction (TAC), while systolic function was
ST:STUDY_SUMMARY                 	preserved. β3AR-expressing hearts displayed enhanced myocardial glucose uptake
ST:STUDY_SUMMARY                 	under stress in absence of increased lactate levels. Instead, metabolomic and
ST:STUDY_SUMMARY                 	metabolic flux analyses in stressed hearts revealed an increase in intermediates
ST:STUDY_SUMMARY                 	of the Pentose-Phosphate Pathway (PPP) in β3AR-Tg, an alternative route of
ST:STUDY_SUMMARY                 	glucose utilization, paralleled with increased transcript levels of
ST:STUDY_SUMMARY                 	NADPH-producing and rate-limiting enzymes of the PPP, without fueling the
ST:STUDY_SUMMARY                 	hexosamine metabolism. The ensuing increased content of NADPH and of reduced
ST:STUDY_SUMMARY                 	glutathione decreased myocyte oxidant stress, while downstream oxidative
ST:STUDY_SUMMARY                 	metabolism assessed by oxygen consumption was preserved with higher glucose
ST:STUDY_SUMMARY                 	oxidation in β3ARTg post-TAC compared to WT, together with increased
ST:STUDY_SUMMARY                 	mitochondrial biogenesis. Unbiased transcriptomics and pathway analysis
ST:STUDY_SUMMARY                 	identified NRF2 (NFE2L2) as upstream transcription factor which was functionally
ST:STUDY_SUMMARY                 	verified in β3AR- expressing cardiac myocytes where its translocation and
ST:STUDY_SUMMARY                 	nuclear activity was dependent on β3AR activation of nitric-oxide synthase
ST:STUDY_SUMMARY                 	(NOS) NO production. Conclusion: Moderate expression of cardiac β3AR, at levels
ST:STUDY_SUMMARY                 	observed in human cardiac myocardium, exerts antioxidant effects through
ST:STUDY_SUMMARY                 	activation of the PPP and NRF2 pathway, thereby preserving myocardial oxidative
ST:STUDY_SUMMARY                 	metabolism, function and integrity under pathophysiological stress.
ST:INSTITUTE                     	UCLouvain
ST:LAST_NAME                     	Dewulf
ST:FIRST_NAME                    	Joseph
ST:ADDRESS                       	Avenue Hippocrate 10
ST:EMAIL                         	joseph.dewulf@uclouvain.be
ST:PHONE                         	027646727
#SUBJECT
SU:SUBJECT_TYPE                  	Mammal
SU:SUBJECT_SPECIES               	Mus musculus
SU:TAXONOMY_ID                   	10090
#FACTORS
#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           	-	013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=022 sample 10 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=023 sample 11 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=024 sample 12 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=025 sample 13 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=026 sample 14 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=027 sample 15 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=028 sample 16 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=029 sample 17 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=030 sample 18 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=031 sample 19 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=032 sample 20 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=033 sample 21 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=034 sample 22 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=035 sample 23 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=036 sample 24 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=037 sample 25 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=038 sample 26 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=039 sample 27 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=040 sample 28 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=041 sample 29 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=042 sample 30 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=043 sample 31 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=044 sample 32 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=013 sample 1 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=014 sample 2 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=015 sample 3 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=016 sample 4 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=019 sample 7 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=020 sample 8 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=021 sample 9 ACNH2O 1_1 AF M1_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	026 sample 14 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=026 sample 14 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	027 sample 15 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=027 sample 15 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	028 sample 16 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=028 sample 16 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	029 sample 17 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=029 sample 17 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	030 sample 18 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=030 sample 18 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	031 sample 19 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=031 sample 19 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	032 sample 20 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=032 sample 20 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	033 sample 21 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=033 sample 21 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	034 sample 22 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=034 sample 22 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	035 sample 23 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=035 sample 23 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	036 sample 24 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=036 sample 24 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	037 sample 25 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=037 sample 25 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	038 sample 26 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=038 sample 26 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	039 sample 27 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=039 sample 27 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	040 sample 28 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=040 sample 28 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	041 sample 29 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=041 sample 29 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	042 sample 30 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=042 sample 30 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	043 sample 31 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=043 sample 31 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	044 sample 32 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=044 sample 32 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	013 sample 1 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=013 sample 1 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	014 sample 2 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=014 sample 2 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	015 sample 3 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=015 sample 3 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	016 sample 4 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=016 sample 4 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	019 sample 7 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=019 sample 7 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	020 sample 8 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=020 sample 8 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	021 sample 9 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=021 sample 9 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	022 sample 10 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=022 sample 10 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	023 sample 11 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=023 sample 11 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	024 sample 12 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=024 sample 12 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	025 sample 13 PREM M2_ MSE_res_pos IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=025 sample 13 PREM M2_ MSE_res_pos IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	026 sample 14 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=026 sample 14  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	027 sample 15 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=027 sample 15  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	028 sample 16 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=028 sample 16  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	029 sample 17 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=029 sample 17  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	030 sample 18 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=030 sample 18  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	031 sample 19 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=031 sample 19  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	032 sample 20 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=032 sample 20  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	033 sample 21 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=033 sample 21  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	034 sample 22 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=034 sample 22  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	035 sample 23 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=035 sample 23  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	036 sample 24 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=036 sample 24  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	037 sample 25 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=037 sample 25  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	038 sample 26 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=038 sample 26  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	039 sample 27 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=039 sample 27  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	040 sample 28 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=040 sample 28  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	041 sample 29 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT TAC 9W	RAW_FILE_NAME(Raw file name)=041 sample 29  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	042 sample 30 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=042 sample 30  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	043 sample 31 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=043 sample 31  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	044 sample 32 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG TAC 9W	RAW_FILE_NAME(Raw file name)=044 sample 32  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	013 sample 1 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=013 sample 1  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	014 sample 2 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=014 sample 2  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	015 sample 3 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=015 sample 3  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	016 sample 4 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=016 sample 4  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	019 sample 7 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=019 sample 7  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	020 sample 8 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=020 sample 8  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	021 sample 9 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=021 sample 9  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	022 sample 10 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=022 sample 10  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	023 sample 11 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:TG Baseline	RAW_FILE_NAME(Raw file name)=023 sample 11  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	024 sample 12 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=024 sample 12  M2_ MSE_res_neg IDC10_ 3ul inj.raw
SUBJECT_SAMPLE_FACTORS           	-	025 sample 13 M2_ MSE_res_neg IDC10_ 3ul inj	Sample source:mouse serum | Condition:WT Baseline	RAW_FILE_NAME(Raw file name)=025 sample 13  M2_ MSE_res_neg IDC10_ 3ul inj.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	Metabolites from serum samples were extracted in microcentrifuge tubes after the
CO:COLLECTION_SUMMARY            	addition of 3 volumes of 100% acetonitrile containing 3 internal standards
CO:COLLECTION_SUMMARY            	[caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic
CO:COLLECTION_SUMMARY            	acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed,
CO:COLLECTION_SUMMARY            	sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10
CO:COLLECTION_SUMMARY            	minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube
CO:COLLECTION_SUMMARY            	and 3 volumes of pure acetonitrile were added to the initial pellet for a second
CO:COLLECTION_SUMMARY            	similar extraction. The combined upper-phases resulting from the two extractions
CO:COLLECTION_SUMMARY            	were mixed and divided into four equal parts. Each part was dried down under a
CO:COLLECTION_SUMMARY            	gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to
CO:COLLECTION_SUMMARY            	analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v),
CO:COLLECTION_SUMMARY            	0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75
CO:COLLECTION_SUMMARY            	(v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the
CO:COLLECTION_SUMMARY            	method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”,
CO:COLLECTION_SUMMARY            	respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to
CO:COLLECTION_SUMMARY            	remove any particulates before being transferred into polypropylene autosampler
CO:COLLECTION_SUMMARY            	vials.
CO:SAMPLE_TYPE                   	Blood (serum)
#TREATMENT
TR:TREATMENT_SUMMARY             	Metabolites from serum samples were extracted in microcentrifuge tubes after the
TR:TREATMENT_SUMMARY             	addition of 3 volumes of 100% acetonitrile containing 3 internal standards
TR:TREATMENT_SUMMARY             	[caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic
TR:TREATMENT_SUMMARY             	acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed,
TR:TREATMENT_SUMMARY             	sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10
TR:TREATMENT_SUMMARY             	minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube
TR:TREATMENT_SUMMARY             	and 3 volumes of pure acetonitrile were added to the initial pellet for a second
TR:TREATMENT_SUMMARY             	similar extraction. The combined upper-phases resulting from the two extractions
TR:TREATMENT_SUMMARY             	were mixed and divided into four equal parts. Each part was dried down under a
TR:TREATMENT_SUMMARY             	gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to
TR:TREATMENT_SUMMARY             	analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v),
TR:TREATMENT_SUMMARY             	0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75
TR:TREATMENT_SUMMARY             	(v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the
TR:TREATMENT_SUMMARY             	method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”,
TR:TREATMENT_SUMMARY             	respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to
TR:TREATMENT_SUMMARY             	remove any particulates before being transferred into polypropylene autosampler
TR:TREATMENT_SUMMARY             	vials. L
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Metabolites from serum samples were extracted in microcentrifuge tubes after the
SP:SAMPLEPREP_SUMMARY            	addition of 3 volumes of 100% acetonitrile containing 3 internal standards
SP:SAMPLEPREP_SUMMARY            	[caffeine-(trimethyl-d9), succinic acid-2,2,3,3-d4 and N-acetyl-aspartic
SP:SAMPLEPREP_SUMMARY            	acid-2,3,3-d3] on one volume of serum. The samples were thoroughly mixed,
SP:SAMPLEPREP_SUMMARY            	sonicated and incubated at -20°C for 30 min, then centrifuged at 10,000g for 10
SP:SAMPLEPREP_SUMMARY            	minutes at 4°C. The upper-phase was collected into a new microcentrifuge tube
SP:SAMPLEPREP_SUMMARY            	and 3 volumes of pure acetonitrile were added to the initial pellet for a second
SP:SAMPLEPREP_SUMMARY            	similar extraction. The combined upper-phases resulting from the two extractions
SP:SAMPLEPREP_SUMMARY            	were mixed and divided into four equal parts. Each part was dried down under a
SP:SAMPLEPREP_SUMMARY            	gentle stream of nitrogen at 30°C and kept frozen until analysis. Prior to
SP:SAMPLEPREP_SUMMARY            	analysis, the samples were reconstituted in [water:acetonitrile, 50:50 (v:v),
SP:SAMPLEPREP_SUMMARY            	0,1% formic acid], [water:acetonitrile, 50:50 (v:v)], [water:acetonitrile, 25:75
SP:SAMPLEPREP_SUMMARY            	(v:v), 0,1% formic acid] and [water:acetonitrile, 25:75 (v:v)] depending of the
SP:SAMPLEPREP_SUMMARY            	method used ,“M1 POS”, “M1 NEG”, “M2 POS” and “M2 NEG”,
SP:SAMPLEPREP_SUMMARY            	respectively. The tubes were centrifuged at 10 000g for 5 minutes at 4°C to
SP:SAMPLEPREP_SUMMARY            	remove any particulates before being transferred into polypropylene autosampler
SP:SAMPLEPREP_SUMMARY            	vials. L
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	M1POS
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity Premier
CH:COLUMN_NAME                   	Waters Acquity Premier HSS T3 1.8um, 2.1 x 100 mm
CH:SOLVENT_A                     	Water, acetic acid 0.1%
CH:SOLVENT_B                     	acetonitrile, acetic acid 0.1%
CH:FLOW_GRADIENT                 	1 min 99%A, 85% over 2 min, 50% over 3min, 5% over 3 min
CH:FLOW_RATE                     	0.5 ml/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Synapt-XS
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Acquisition: Masslynx The separation columns, operated at 40°C, were an Acquity
MS:MS_COMMENTS                   	Premier HSS T3 column 1,8 µm, 2,1 x 100 mm (Waters) and an Acquity Premier BEH
MS:MS_COMMENTS                   	Amide 1,8 µm, 2,1 x 100 mm (Waters), for reverse phase and HILIC
MS:MS_COMMENTS                   	chromatography, respectively. Mass spectrometry data was acquired in profile
MS:MS_COMMENTS                   	(continuum) format over the mass range of m/z 50-1200 using a Waters Synapt XS
MS:MS_COMMENTS                   	high resolution Q-TOF mass spectrometer set in MSE resolution mode (scan time
MS:MS_COMMENTS                   	0,1 sec, the low and high trap collision energy were 4V and a ramp between 20
MS:MS_COMMENTS                   	and 50 V, respectively). A dual electrospray ionization (ESI) source was used in
MS:MS_COMMENTS                   	positive or negative mode. Capillary and sampling cone, voltages were set to 1
MS:MS_COMMENTS                   	kV and 30 V or 1 kV and 25 V in positive and negative mode, respectively. Source
MS:MS_COMMENTS                   	temperature was set to 150 °C and desolvation temperature to 600 °C (550°C in
MS:MS_COMMENTS                   	negative ESI). Gas flow rates were set at 1200 L/h for the desolvation gas (1100
MS:MS_COMMENTS                   	L/h in negative ESI) and 50 L/h for the cone gas. Acquisition of leucine
MS:MS_COMMENTS                   	enkephalin infused at 10ul/min through a lockspray probe allowed a real time
MS:MS_COMMENTS                   	mass correction (30 sec scan intervals). Data processing: Raw LC-MS data were
MS:MS_COMMENTS                   	imported and processed in Progenesis QI software (Nonlinear Dynamics), which
MS:MS_COMMENTS                   	performed chromatogram alignment, peak picking, ion deconvolution,
MS:MS_COMMENTS                   	normalization, peak annotation and statistical analysis. Thresholds of area
MS:MS_COMMENTS                   	under the curve > 1000 were set and yielded the following number of detected
MS:MS_COMMENTS                   	metabolites: for Control WT vs Control TG (M1 neg : 1886 ; M1 pos : 3881 ; M2
MS:MS_COMMENTS                   	neg : 785 ; M2 pos : 2411) TAC 9W WT vs Control TG (M1 neg : 2022 ; M1 pos :
MS:MS_COMMENTS                   	3690 ; M2 neg : 905 ; M2 pos : 2405) and Control WT vs TAC WT (M1 neg : 2165 ;
MS:MS_COMMENTS                   	M1 pos : 4068 ; M2 neg : 944 ; M2 pos : 2448). Volcano plot and FDR threshold of
MS:MS_COMMENTS                   	significance were performed on Graphpad Prism.
MS:MS_RESULTS_FILE               	ST003507_AN005758_Results.txt	UNITS:arbitrary unit	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END