#METABOLOMICS WORKBENCH kevin.cho@wustl.edu_20240409_091613 DATATRACK_ID:4766 STUDY_ID:ST003162 ANALYSIS_ID:AN005188 PROJECT_ID:PR001967
VERSION             	1
CREATED_ON             	April 10, 2024, 6:57 pm
#PROJECT
PR:PROJECT_TITLE                 	Loss of dihydroceramide desaturase drives neurodegeneration by disrupting
PR:PROJECT_TITLE                 	endoplasmic reticulum and lipid droplet homeostasis in glial cells
PR:PROJECT_TYPE                  	Untargeted Metabolomics and Lipidomics
PR:PROJECT_SUMMARY               	Dihydroceramide desaturases convert dihydroceramides to ceramides, the
PR:PROJECT_SUMMARY               	precursors of all complex sphingolipids. Reduction of DEGS1 dihydroceramide
PR:PROJECT_SUMMARY               	desaturase function causes pediatric neurodegenerative disorder hypomyelinating
PR:PROJECT_SUMMARY               	leukodystrophy-18 (HLD-18). We discovered that infertile crescent (ifc), the
PR:PROJECT_SUMMARY               	Drosophila DEGS1 homolog, is expressed primarily in glial cells to promote CNS
PR:PROJECT_SUMMARY               	development by guarding against neurodegeneration. Loss of ifc causes massive
PR:PROJECT_SUMMARY               	dihydroceramide accumulation and severe morphological defects in cortex glia,
PR:PROJECT_SUMMARY               	including endoplasmic reticulum (ER) expansion, failure of neuronal
PR:PROJECT_SUMMARY               	ensheathment, and lipid droplet depletion. RNAi knockdown of the upstream
PR:PROJECT_SUMMARY               	ceramide synthase schlank in glia of ifc mutants rescues ER expansion,
PR:PROJECT_SUMMARY               	suggesting dihydroceramide accumulation in the ER drives this phenotype. RNAi
PR:PROJECT_SUMMARY               	knockdown of ifc in glia but not neurons drives neuronal cell death, suggesting
PR:PROJECT_SUMMARY               	that ifc function in glia promotes neuronal survival. Our work identifies glia
PR:PROJECT_SUMMARY               	as the primary site of disease progression in HLD-18 and may inform on juvenile
PR:PROJECT_SUMMARY               	forms of ALS, which also feature elevated dihydroceramide levels.
PR:INSTITUTE                     	Washington University in St. Louis
PR:DEPARTMENT                    	Genetics, Medicine, Chemistry
PR:LABORATORY                    	Skeath and Patti Laboratories
PR:LAST_NAME                     	Cho
PR:FIRST_NAME                    	Kevin
PR:ADDRESS                       	1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA
PR:EMAIL                         	kevin.cho@wustl.edu
PR:PHONE                         	314-935-8813
#STUDY
ST:STUDY_TITLE                   	Loss of dihydroceramide desaturase drives neurodegeneration by disrupting
ST:STUDY_TITLE                   	endoplasmic reticulum and lipid droplet homeostasis in glial cells
ST:STUDY_TYPE                    	Untargeted Metabolomics & Lipidomics
ST:STUDY_SUMMARY                 	Dihydroceramide desaturases convert dihydroceramides to ceramides, the
ST:STUDY_SUMMARY                 	precursors of all complex sphingolipids. Reduction of DEGS1 dihydroceramide
ST:STUDY_SUMMARY                 	desaturase function causes pediatric neurodegenerative disorder hypomyelinating
ST:STUDY_SUMMARY                 	leukodystrophy-18 (HLD-18). We discovered that infertile crescent (ifc), the
ST:STUDY_SUMMARY                 	Drosophila DEGS1 homolog, is expressed primarily in glial cells to promote CNS
ST:STUDY_SUMMARY                 	development by guarding against neurodegeneration. Loss of ifc causes massive
ST:STUDY_SUMMARY                 	dihydroceramide accumulation and severe morphological defects in cortex glia,
ST:STUDY_SUMMARY                 	including endoplasmic reticulum (ER) expansion, failure of neuronal
ST:STUDY_SUMMARY                 	ensheathment, and lipid droplet depletion. RNAi knockdown of the upstream
ST:STUDY_SUMMARY                 	ceramide synthase schlank in glia of ifc mutants rescues ER expansion,
ST:STUDY_SUMMARY                 	suggesting dihydroceramide accumulation in the ER drives this phenotype. RNAi
ST:STUDY_SUMMARY                 	knockdown of ifc in glia but not neurons drives neuronal cell death, suggesting
ST:STUDY_SUMMARY                 	that ifc function in glia promotes neuronal survival. Our work identifies glia
ST:STUDY_SUMMARY                 	as the primary site of disease progression in HLD-18 and may inform on juvenile
ST:STUDY_SUMMARY                 	forms of ALS, which also feature elevated dihydroceramide levels.
ST:INSTITUTE                     	Washington University in St. Louis
ST:DEPARTMENT                    	Genetics, Medicine, Chemistry
ST:LABORATORY                    	Skeath and Patti Laboratories
ST:LAST_NAME                     	Cho
ST:FIRST_NAME                    	Kevin
ST:ADDRESS                       	1 Brookings Drive, Campus Box 1134, St. Louis, MO, 63130, USA
ST:EMAIL                         	kevin.cho@wustl.edu
ST:PHONE                         	314-935-8813
#SUBJECT
SU:SUBJECT_TYPE                  	Insect
SU:SUBJECT_SPECIES               	Drosophila melanogaster
SU:TAXONOMY_ID                   	7227
#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           	-	WL_RPLC_POS_ifc_1	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_ifc_2	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_ifc_3	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_ifc_4	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_ifc_5	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_ifc_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_WT_1	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_WT_2	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_WT_3	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_WT_4	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_RPLC_POS_WT_5	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=WL_RPLC_POS_WT_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_ifc_1	Genotype:Knockout | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_ifc_2	Genotype:Knockout | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_ifc_3	Genotype:Knockout | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_ifc_4	Genotype:Knockout | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_ifc_5	Genotype:Knockout | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_ifc_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_WT_1	Genotype:Wild-type | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_WT_2	Genotype:Wild-type | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_WT_3	Genotype:Wild-type | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_WT_4	Genotype:Wild-type | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_RPLC_POS_WT_5	Genotype:Wild-type | Sample source:CNS	Chromatography=RPLC; Polarity=Positive; RAW_FILE_NAME(Raw file name)=CNS_RPLC_POS_WT_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_ifc_1	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_ifc_2	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_ifc_3	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_ifc_4	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_ifc_5	Genotype:Knockout | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_ifc_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_WT_1	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_WT_2	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_WT_3	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_WT_4	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	WL_HILIC_NEG_WT_5	Genotype:Wild-type | Sample source:Whole Larvae	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=WL_HILIC_NEG_WT_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_ifc_1	Genotype:Knockout | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_ifc_2	Genotype:Knockout | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_ifc_3	Genotype:Knockout | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_ifc_4	Genotype:Knockout | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_ifc_5	Genotype:Knockout | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_ifc_5.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_WT_1	Genotype:Wild-type | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_1.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_WT_2	Genotype:Wild-type | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_2.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_WT_3	Genotype:Wild-type | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_3.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_WT_4	Genotype:Wild-type | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_4.mzML
SUBJECT_SAMPLE_FACTORS           	-	CNS_HILIC_NEG_WT_5	Genotype:Wild-type | Sample source:CNS	Chromatography=HILIC; Polarity=Negative; RAW_FILE_NAME(Raw file name)=CNS_HILIC_NEG_WT_5.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Whole larvae and dissected CNS
CO:SAMPLE_TYPE                   	Larvae
#TREATMENT
TR:TREATMENT_SUMMARY             	Untargeted lipidomics analysis was conducted on whole larva and dissected CNS of
TR:TREATMENT_SUMMARY             	wild type and ifc-/- mutants at the late-third instar stage. Five replicates
TR:TREATMENT_SUMMARY             	were prepared for each set of experiments. For whole larvae, at least 15 larvae
TR:TREATMENT_SUMMARY             	of each genotype were used for each replicate. For the dissected CNS, at least
TR:TREATMENT_SUMMARY             	50 wild type and 60 ifc-/- CNS were used per replicate. Immediately following
TR:TREATMENT_SUMMARY             	collection or dissection, larvae and the dissected CNS were flash frozen in
TR:TREATMENT_SUMMARY             	liquid nitrogen and placed at -80°C.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Whole larvae and CNS samples were extracted utilizing a tissue homogenizer,
SP:SAMPLEPREP_SUMMARY            	employing a solvent mixture of acetonitrile, methanol, and water in a 2:2:1
SP:SAMPLEPREP_SUMMARY            	ratio, using 40 µL per mg of wet weight. The samples were then stored at -20°C
SP:SAMPLEPREP_SUMMARY            	overnight. Following centrifugation at 14,000 x g for 10 minutes at 4°C, the
SP:SAMPLEPREP_SUMMARY            	supernatant was transferred into an LC/MS vial and subsequently stored at -80°C
SP:SAMPLEPREP_SUMMARY            	until LC/MS analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish Flex UHPLC Systems
CH:COLUMN_NAME                   	HILICON iHILIC-(P) Classic (100 x 2.1mm,5um)
CH:SOLVENT_A                     	water/acetonitrile (95/5); 20 mM ammonium bicarbonate; 0.1% ammonium hydroxide;
CH:SOLVENT_A                     	2.5 μM medronic acid
CH:SOLVENT_B                     	acetonitrile/water (95/5)
CH:FLOW_GRADIENT                 	0–1 min: 90% B, 1–12 min: 90-35% B, 12–12.5 min: 35-25% B, 12.5–14.5
CH:FLOW_GRADIENT                 	min: 25% B
CH:FLOW_RATE                     	250 uL/min
CH:COLUMN_TEMPERATURE            	45
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap ID-X tribrid
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Data were collected with the following MS source settings: spray voltage, -2.8
MS:MS_COMMENTS                   	kV; sheath gas, 50; auxiliary gas, 10; sweep gas, 1; ion transfer tube
MS:MS_COMMENTS                   	temperature, 300°C; vaporizer temperature, 200°C; mass range, 67 – 1000 Da;
MS:MS_COMMENTS                   	resolution, 120,000; maximum injection time, 200 ms; isolation window, 1.6 Da.
MS:MS_COMMENTS                   	XCMS and Skyline software were used for data processing
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	CNS_HILIC_NEG_WT_1	CNS_HILIC_NEG_WT_2	CNS_HILIC_NEG_WT_3	CNS_HILIC_NEG_WT_4	CNS_HILIC_NEG_WT_5	CNS_HILIC_NEG_ifc_1	CNS_HILIC_NEG_ifc_2	CNS_HILIC_NEG_ifc_3	CNS_HILIC_NEG_ifc_4	CNS_HILIC_NEG_ifc_5
Factors	Genotype:Wild-type | Sample source:CNS	Genotype:Wild-type | Sample source:CNS	Genotype:Wild-type | Sample source:CNS	Genotype:Wild-type | Sample source:CNS	Genotype:Wild-type | Sample source:CNS	Genotype:Knockout | Sample source:CNS	Genotype:Knockout | Sample source:CNS	Genotype:Knockout | Sample source:CNS	Genotype:Knockout | Sample source:CNS	Genotype:Knockout | Sample source:CNS
Glucose	74696	179970	207611	142268	110533	285615	334420	199017	405114	460725
Glucose-6-phosphate	10462747	8898463	7817362	5608536	3319351	8926097	5340995	6592602	3839139	11148568
Fructose-bisphosphate	4769995	3639578	3524429	1766953	1181870	1829980	1201933	2593294	913269	2745753
DHAP	3668858	3187430	2541948	2480262	1786925	2512405	1582788	1876758	863240	2856584
2/3-PG	10117220	7404940	7975729	8637683	4896285	7776982	4580916	6725154	2229932	7389270
Phosphoenolpyruvate	1350545	1256355	1105873	2056861	1057636	1867847	819847	1066770	456797	1155046
Pyruvate	6781792	6853737	7733112	7753924	4939750	5194759	2186314	3108875	2199156	4470578
Lactate	163367072	162557424	178863696	162612688	55214372	142212080	116097240	123674496	108373640	220292624
Ribose-5-phosphate	279939	222457	238256	248302	116767	178134	115994	89377	46108	230322
Xylulose-5-phosphate	1002874	751677	632660	584617	294921	173909	164733	249856	128977	272931
Sedoheptulose 7-phosphate	701029	516959	455236	374832	225028	666551	392189	479461	322631	730300
Citrate	14376626	10081685	12763893	11270391	8440789	8679889	7003505	11779377	6850207	7893539
Aconitate	432687	305538	412199	459274	232611	224623	182380	146754	148847	244208
a-KG	3552646	3217245	6002153	4809215	4451411	1581942	1938666	517468	2804987	535148
Succinate	4849468	4398069	5584810	4453286	3717677	3404759	3040595	2728796	3745989	3313411
Fumarate	4968113	3882456	4069848	4220959	2453214	1482555	1152324	980629	1287951	1145254
Malate	92474904	74137832	78688448	79680816	51784788	31037792	27295614	22730766	32364764	27176226
Glutamate	72897584	61447276	62316960	62789948	38095816	29617358	24109804	20739730	20510334	31097440
Glutamine	9216221	8321454	9242683	4714363	3387664	9234541	7893601	8010228	5861898	11096252
Aspartate	18008528	15506260	13777500	17244506	9513131	9826667	8109509	10005007	9324621	10619394
Alanine	18446084	16033727	16917788	17420428	11125637	10984710	9291824	8230299	9420786	12983297
Glycine	3459788	2995372	2913461	2960821	2244183	3135420	2482299	1827466	2461698	2770863
Valine	6420958	4918644	5305802	4611946	3072850	4301667	3218734	3148612	3093621	4647418
Leucine	12869739	10067751	11021671	10085213	6430167	9617550	7535678	6468857	6041789	9452037
Isoleucine	5630789	4578674	4218202	4046444	2523233	3752941	3160560	3698076	2820690	4142319
Asparagine	7054756	4947224	5668789	4609133	3099465	4413688	3626815	3726911	3313147	4281615
Proline	14182377	12749668	12712447	10434846	7056130	8217975	7168406	6767594	5561083	8467954
Histidine	55436208	38472448	45683116	37391596	29732840	53078336	41413600	36548380	42731448	44402500
Methionine	400898	423117	379857	652420	330819	340875	273561	271239	221745	506647
Lysine	177161	152771	177257	139709	115717	184762	152845	222214	155389	147254
Arginine	9149549	7235640	7771671	6510176	4164172	9306020	7276095	14209337	5497869	9729785
Phenylalanine	10905424	7887443	8643994	7332766	5210003	8391279	6082753	5203893	5135778	6556145
Serine	5366600	3876879	4350332	3517178	2631896	3904262	2791387	2904754	2611120	3365434
Threonine	4532314	3588058	4405262	3965794	2751471	3291358	2383681	2060625	2410975	3057937
Tyrosine	9175983	7685924	7635100	8866497	5547080	4406173	4348334	3494135	3181079	5037070
Glycerol 3-phosphate	14228118	11544128	9501583	14799665	7757904	9946407	6767875	6576690	3850885	10728683
Adenine	2170599	2590831	2763518	3583098	1206636	2887727	1983672	1325210	1357729	2739640
Uracil	8391266	7621532	7108142	7973626	4529043	2567129	1838600	1989940	1384695	3192395
Guanosine	133224	187694	83392	191482	30966	100744	103260	59037	69043	207606
Uridine	3084498	2151790	2006214	1173855	645614	559785	509376	548705	433362	1173459
Cytidine	1624881	1594032	1159073	1900164	632334	336928	256765	291163	231967	549292
Glutathione, reduced	30069660	22450640	21579182	27472310	15412513	20214324	16281416	17832258	9530146	21706466
Glutathione, oxidized	64590	76752	55044	127622	46612	51800	55089	179256	70863	66812
UMP	246253	292916	184313	244008	86083	304412	218946	261125	175675	302227
UDP	222518	250796	149220	143697	100402	166153	128919	160560	124556	220495
UTP	64437	24777	30141	8428	18962	27457	15570	40140	8716	31633
AMP	11928166	12923707	8011402	12620946	4089593	9456784	7538137	6868374	5057153	11322906
ADP	6917115	5697804	3918895	5352337	2158835	3667045	3446770	3551350	2604289	4244692
ATP	1614866	961505	990525	509732	557205	477663	504099	994607	357856	554265
NAD+	2270037	1874030	1763841	1965699	932544	2117275	1777953	1454061	1170756	2235945
NADH	1196812	569495	539429	342399	211488	572599	392534	475828	179454	769694
NADP+	26965	25502	6638	11508	2899	14760	15160	37266	17483	21961
CMP	109463	144018	62337	136551	38194	77652	64613	52812	36873	81030
CDP	186815	184676	116134	124079	68092	76406	76248	104435	48414	92743
CDP-Choline	176103	125598	144142	80872	61232	64836	52087	65766	33865	72626
FAD	605012	560628	514977	488946	200263	581107	414942	493082	233052	646078
GMP	862530	824882	420832	786439	261324	377160	324169	330604	251194	525127
GDP	268130	299310	221860	183622	132200	154427	156973	163766	135279	189150
dGTP	1616778	963589	992979	509732	560080	477670	504107	994592	358254	554273
UDP-glucose / UDP-galatose	1940225	1432762	1352815	1140533	713504	1055090	747804	1009512	553516	1376280
Uridine 5'-diphosphoglucuronic acid	52738	47016	39415	36466	12228	46113	37799	131237	24483	51081
UDP-N-acetylglucosamine	2769260	2043535	2080915	1873956	1112215	1469386	1258949	1477529	889110	2091926
Glucosamine-6-phosphate	891093	558200	606170	588668	391188	917027	935100	630629	674126	1147025
GlcNAc-Phosphate	257271	195743	191753	194855	108367	203878	191827	189138	110732	253865
S-Adenosylhomocysteine	447524	267359	230862	441766	187489	213612	152797	202847	103845	234831
S-Adenosylmethionine	1307916	962768	958909	858074	579742	649950	526820	505056	299540	706814
Carnitine	174012832	124040536	109210808	105368328	58189568	74499152	64668264	78192656	55985516	95213608
3-Indoleacrylic acid	49177744	34631228	37051248	32638162	25186510	43740824	34041252	30559054	28708754	38940256
Acetyl-Lysine	140550	376874	613263	668033	93733	715909	443792	221807	349601	584344
N,N-Dimethylarginine	386399	359457	284960	339507	128340	318945	243109	112698	127301	410753
Acetylcarnitine	270554848	215202432	240633424	217319808	132356800	185426928	152079488	135105520	85816520	192298144
Glu-gln	1994686	1650913	1945404	1722750	1035031	901941	735006	849137	378636	975247
Methyladenosine	271705	254633	223983	217357	134891	185874	117470	102463	48366	221683
Cysteine-glutathione disulfide	277717	245956	319839	418672	88807	180520	176579	272320	189422	219541
2KIVA	1032692	1013943	1034906	1010798	642026	947922	517305	2127795	890938	533158
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
Glucose
Glucose-6-phosphate
Fructose-bisphosphate
DHAP
2/3-PG
Phosphoenolpyruvate
Pyruvate
Lactate
Ribose-5-phosphate
Xylulose-5-phosphate
Sedoheptulose 7-phosphate
Citrate
Aconitate
a-KG
Succinate
Fumarate
Malate
Glutamate
Glutamine
Aspartate
Alanine
Glycine
Valine
Leucine
Isoleucine
Asparagine
Proline
Histidine
Methionine
Lysine
Arginine
Phenylalanine
Serine
Threonine
Tyrosine
Glycerol 3-phosphate
Adenine
Uracil
Guanosine
Uridine
Cytidine
Glutathione, reduced
Glutathione, oxidized
UMP
UDP
UTP
AMP
ADP
ATP
NAD+
NADH
NADP+
CMP
CDP
CDP-Choline
FAD
GMP
GDP
dGTP
UDP-glucose / UDP-galatose
Uridine 5'-diphosphoglucuronic acid
UDP-N-acetylglucosamine
Glucosamine-6-phosphate
GlcNAc-Phosphate
S-Adenosylhomocysteine
S-Adenosylmethionine
Carnitine
3-Indoleacrylic acid
Acetyl-Lysine
N,N-Dimethylarginine
Acetylcarnitine
Glu-gln
Methyladenosine
Cysteine-glutathione disulfide
2KIVA
METABOLITES_END
#END