#METABOLOMICS WORKBENCH alanyl_20250326_085903 DATATRACK_ID:5786 STUDY_ID:ST003826 ANALYSIS_ID:AN007478 PROJECT_ID:PR002390
VERSION             	1
CREATED_ON             	March 30, 2025, 2:56 pm
#PROJECT
PR:PROJECT_TITLE                 	In vivo CRISPR screen identifies copper metabolism as a vulnerability in acute
PR:PROJECT_TITLE                 	lymphoblastic leukemia
PR:PROJECT_SUMMARY               	The nutrient-sparse cerebrospinal fluid (CSF) poses a significant challenge to
PR:PROJECT_SUMMARY               	spreading cancer cells. Despite this challenge, leukemia often spreads to the
PR:PROJECT_SUMMARY               	CSF and represents a significant clinical complication. To uncover nutritional
PR:PROJECT_SUMMARY               	dependencies of leukemia cells in the CSF that could be targeted
PR:PROJECT_SUMMARY               	therapeutically, we conducted an in vivo targeted CRISPR screen in a xenograft
PR:PROJECT_SUMMARY               	model of leukemia. We found that SLC31A1, the primary cell surface copper
PR:PROJECT_SUMMARY               	importer, is a genetic dependency of leukemia in both the central nervous system
PR:PROJECT_SUMMARY               	as well as in the hematopoietic organs. Perturbation of copper metabolism leads
PR:PROJECT_SUMMARY               	to complex IV deficiency, perturbed nucleotide metabolism and slowed leukemia
PR:PROJECT_SUMMARY               	cell proliferation. Furthermore, nutritional copper depletion reduced cancer
PR:PROJECT_SUMMARY               	progression in cell line based and patient-derived xenograft models of leukemia.
PR:PROJECT_SUMMARY               	Copper thus appears to be an actionable micronutrient in leukemia.
PR:INSTITUTE                     	Boston Children's Hospital
PR:DEPARTMENT                    	Pathology
PR:LABORATORY                    	Naama Kanarek
PR:LAST_NAME                     	Wong
PR:FIRST_NAME                    	Alan
PR:ADDRESS                       	300 Longwood Avenue, Boston, MA, 02115, USA
PR:EMAIL                         	alan.wong@childrens.harvard.edu
PR:PHONE                         	(617) 355-7433
PR:FUNDING_SOURCE                	NCI 1R01CA282477-01A1
#STUDY
ST:STUDY_TITLE                   	Polar metabolite profiling of TTM-treated SEM cells
ST:STUDY_SUMMARY                 	To investigate the effects of ammonium tetrathiomolybdate (TTM) (a copper
ST:STUDY_SUMMARY                 	chelator) on steady state metabolite levels, SEM cells were treated for 2, 6 or
ST:STUDY_SUMMARY                 	24 hours with 100uM TTM; control cells received DMSO for 2 hours. This
ST:STUDY_SUMMARY                 	experiment revealed that TTM treatment significantly perturbed nucleotide
ST:STUDY_SUMMARY                 	levels.
ST:INSTITUTE                     	Boston Childrens Hospital
ST:LAST_NAME                     	Wong
ST:FIRST_NAME                    	Alan
ST:ADDRESS                       	300 Longwood Avenue
ST:EMAIL                         	alan.wong@childrens.harvard.edu
ST:PHONE                         	(617) 355-7433
ST:NUM_GROUPS                    	4
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
SU:CELL_STRAIN_DETAILS           	SEM leukemia cells
#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           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1103	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1103.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1104	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1104.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1105	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1105.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1106	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1106.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1107	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1107.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1108	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1108.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1109	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1109.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1110	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1110.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1111	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1111.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1112	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1112.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1113	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1113.raw
SUBJECT_SAMPLE_FACTORS           	-	20240412_QE2_HILIC_TTM_timecourse_AYW1114	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2	RAW_FILE_NAME(raw file name)=20240412_QE2_HILIC_TTM_timecourse_AYW1114.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	One and a half million cells from culture were collected via centrifugation for
CO:COLLECTION_SUMMARY            	20 seconds at 18,000xG at 4C, washed once with ice-cold 0.9% NaCl, and collected
CO:COLLECTION_SUMMARY            	via centrifugation for 20 seconds at 18,000xG at 4C. Cells were cultured in
CO:COLLECTION_SUMMARY            	RPMI-1640 with 10% FBS and penicillin/streptomycin in a 37C incubator with 5%
CO:COLLECTION_SUMMARY            	CO2.
CO:SAMPLE_TYPE                   	Leukemia cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were treated for up to 24 hours with vehicel (DMSO) or 100uM TTM. Culture
TR:TREATMENT_SUMMARY             	media was RPMI-1640 containing 10% FBS and penicillin and streptomycin in 6-well
TR:TREATMENT_SUMMARY             	plates. Cells were seeded at 1.5 million/mL in 4mL.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Cell pellet was resuspended in 400uL of 100% LC-MS grade methanol supplemented
SP:SAMPLEPREP_SUMMARY            	with isotopically-labelled amino acid standards [Cambridge Isotope Laboratories,
SP:SAMPLEPREP_SUMMARY            	MSK-A2-1.2], aminopterin, and reduced glutathione standard [Cambridge Isotope
SP:SAMPLEPREP_SUMMARY            	Laboratories, CNLM-6245-10]). with repeated pipetting up and down and vortexing
SP:SAMPLEPREP_SUMMARY            	for 10 seconds. Then, 100uL of LCMS-grade water containing 125 mM Ammonium
SP:SAMPLEPREP_SUMMARY            	Acetate, 10 mM Na-Ascorbate, and 7.9 mg/mL 5,5-dithio-bis-(2-nitrobenzoic acid
SP:SAMPLEPREP_SUMMARY            	(Ellman's reagent) was added, and the sample vortex for another 10 seconds.
SP:SAMPLEPREP_SUMMARY            	Samples were then centrifuged for 10 minutes at 18,000 g to pellet cell debris.
SP:SAMPLEPREP_SUMMARY            	The supernatant was transferred to a new tube and dried on ice using a liquid
SP:SAMPLEPREP_SUMMARY            	nitrogen dryer. The dried metabolites were then resuspended in 30uL and 2uL was
SP:SAMPLEPREP_SUMMARY            	injected.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	2 μL of each sample was injected into a ZIC-pHILIC 150 x 2.1 mm (5 μm particle
CH:CHROMATOGRAPHY_SUMMARY        	size) column (EMD Millipore) operated on a Vanquish™ Flex UHPLC system (Thermo
CH:CHROMATOGRAPHY_SUMMARY        	Fisher Scientific). Chromatographic separation was achieved using the following
CH:CHROMATOGRAPHY_SUMMARY        	conditions: buffer A was 95% acetonitrile and 5% 20 mM ammonium carbonate, 0.1%
CH:CHROMATOGRAPHY_SUMMARY        	ammonium hydroxide in water; buffer B was 95% 20 mM ammonium carbonate, 0.1%
CH:CHROMATOGRAPHY_SUMMARY        	ammonium hydroxide in water and 5% acetonitrile. Gradient conditions used were:
CH:CHROMATOGRAPHY_SUMMARY        	0-20 min: linear gradient from 16.6% to 83.4% B; 20-24 min: hold at 83.4% B;
CH:CHROMATOGRAPHY_SUMMARY        	24-24.1 min: from 83.4% to 16.6% B; 24.1-32 min: hold at 20% B at 0.150 mL/min
CH:CHROMATOGRAPHY_SUMMARY        	flow rate. The column oven and autosampler tray were held at 25 °C and 4 °C,
CH:CHROMATOGRAPHY_SUMMARY        	respectively.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	SeQuant ZIC-HILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	95% acetonitrile; 5% water with 20mM ammonium carbonate; 0.1% ammonium hydroxide
CH:SOLVENT_B                     	95% water with 20mM ammonium carbonate; 0.1% ammonium hydroxide; 5% acetonitrile
CH:FLOW_GRADIENT                 	Gradient conditions used were: 0-20 min: linear gradient from 16.6% to 83.4% B;
CH:FLOW_GRADIENT                 	20-24 min: hold at 83.4% B; 24-24.1 min: from 83.4% to 16.6% B; 24.1-32 min:
CH:FLOW_GRADIENT                 	hold at 20% B at 0.150 mL/min flow rate. The column oven and autosampler tray
CH:FLOW_GRADIENT                 	were held at 25 °C and 4 °C, respectively.
CH:FLOW_RATE                     	0.15 mL/min
CH:COLUMN_TEMPERATURE            	25
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	UNSPECIFIED
MS:MS_COMMENTS                   	MS data acquisition was performed using a QExactive benchtop orbitrap mass
MS:MS_COMMENTS                   	spectrometer equipped with an Ion Max source and a HESI II probe (Thermo Fisher
MS:MS_COMMENTS                   	Scientific) and polarity switching was used. Four scans were used: full scans in
MS:MS_COMMENTS                   	both positive and negative ionization mode in a range of m/z = 70–1000, with
MS:MS_COMMENTS                   	the resolution set at 70,000, the AGC target at 1 × 10e6, and the maximum
MS:MS_COMMENTS                   	injection time (Max IT) at 20 msec from 0-20 minutes. A third scan in the
MS:MS_COMMENTS                   	negative mode was used with range of m/z = 220-700 from 0-20 minutes and the
MS:MS_COMMENTS                   	same resolution, AGC settings with 30ms Max IT. Lastly, a targeted-SIM scan was
MS:MS_COMMENTS                   	added with a resolution of 35k, AGC target 1e5, and max IT 20ms, isolation
MS:MS_COMMENTS                   	window = 1.0 m/z, with an inclusion m/z of 503.0552 (corresponding to
MS:MS_COMMENTS                   	Ellman-derivatized glutathione). Tune file parameters were: spray voltage =
MS:MS_COMMENTS                   	3.5kV, capillary temperature = 320C, S-lens RF = 50, auxillary gas temperature =
MS:MS_COMMENTS                   	350C.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Normalized peak area
MS_METABOLITE_DATA_START
Samples	20240412_QE2_HILIC_TTM_timecourse_AYW1103	20240412_QE2_HILIC_TTM_timecourse_AYW1104	20240412_QE2_HILIC_TTM_timecourse_AYW1105	20240412_QE2_HILIC_TTM_timecourse_AYW1106	20240412_QE2_HILIC_TTM_timecourse_AYW1107	20240412_QE2_HILIC_TTM_timecourse_AYW1108	20240412_QE2_HILIC_TTM_timecourse_AYW1109	20240412_QE2_HILIC_TTM_timecourse_AYW1110	20240412_QE2_HILIC_TTM_timecourse_AYW1111	20240412_QE2_HILIC_TTM_timecourse_AYW1112	20240412_QE2_HILIC_TTM_timecourse_AYW1113	20240412_QE2_HILIC_TTM_timecourse_AYW1114
Factors	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:24	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:6	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	Sample source:SEM leukemia cells | Treatment:TTM | Treatment time:2	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2	Sample source:SEM leukemia cells | Treatment:DMSO | Treatment time:2
2-aminobutyric_acid	35404357.56	31274235.11	33649214.47	23403473.23	24620825.18	24237116.81	27318879.21	25154448.12	21547629.51	25144620.61	27736730.62	26713993.46
3-phosphoglycerate	2511991.287	4695640.54	3640658.1	10813010.03	10895733.05	11121882.98	5934776.846	5567807.517	5117207.117	3895725.944	4769291.173	4187751.633
Acetyl.CoA	3141127.998	3594215.652	2298244.944	7885317.991	8355353.31	8067454.309	8136982.477	8662543.042	7621675.376	7848357.225	8834748.467	7658702.329
acetylcarnitine	242078915.8	239470103.5	223586382.7	301106644.3	297551392.9	292574831.7	318807249.1	318456017.6	321430368.4	331789149	327082286.1	303213685.7
ADP	84124507.17	86867896.2	74723427.95	111176283.8	102341017.4	97978009.47	108724146.7	108991746	100263044	105421070.9	122694011.2	114986314.1
alanine	91744049.74	80715669.1	87929339.29	72208787.79	74893183.71	74424635.01	77560155.39	80588331.87	77567599.96	87755281.62	79403489.01	80308093.31
Alpha.Ketoglutarate	116042273.8	101351364.2	116173011.3	98889556.12	105026742	100454010.7	82601177.21	77343155.78	62204353.36	69147048.02	63611654.19	70774041.99
AMP	36319550.85	34091343.73	34266598.8	24212201.15	23884592.27	24270000.98	22451428.32	22245584.84	23824977.79	22834482.9	24504141.13	26122463.5
arginine	1870062174	2059753010	1888812809	1652619627	1550150771	1635707696	1618788269	1667925371	1597505239	1733092132	1778075184	1928819713
argininosuccinate	10269467.96	10772279.76	11460933.7	12995081.45	13573621.13	13417623.35	8131640.443	7592204.143	8354635.332	7314142.338	6999168.344	6911323.006
asparagine	101334804	94873151.71	103306566.1	156701200.3	151104433.9	156238409.2	167517163.9	165925583.9	165324283.8	174919774.2	170738058.7	172606568.2
aspartate	42677346.28	37787319.65	38523013.23	89832086.86	90627452.46	94947172.15	138056008.4	133679598.4	141337236.6	140211047.1	136286214.5	132812913.9
asymmetric.dimethylarginine	11175708.39	12655734.09	10587751.63	6233837.784	6160886.649	6967411.402	6473776.08	7274659.563	6384847.251	7028461.081	7197592.425	7574201.516
ATP	268273151.4	347763775.3	221813291.7	764475629.8	571026843.4	537901608.7	638015754.7	728268313.7	492273025.2	513577219.3	762086963.4	647151765.1
Butyrylcarnitine	46482754.49	46967601.59	53564909.87	102198314.9	122853836.8	119370093.1	120600628.4	135504302.5	177176775.4	137236044.8	110094478.8	140338144.3
carnitine	325825885.6	336705979.1	333609378.2	344479787.1	384229032.5	379230664.4	437253552.7	401708665.1	419370127.1	423270145.5	385816756.4	404605683.1
cis.aconitate	65042245.61	63053365.37	64890239.75	48754425.98	49080319.42	54737538.75	41789846.85	33293482	36485203.54	32127416.2	39067166.79	43121594.49
citrate	890576434	686427750.5	725314970.5	415701870.9	441868040.7	413419714.9	513263280	429983804.1	494439070.4	606584487.2	482117678.6	496350034.3
citrulline	22645940.84	23203758.92	23101714.85	22752635.34	23500037.77	25130774.93	24207040.55	27122500.71	24221765.37	25975356.41	24631614.88	26192279.25
creatine	293423349.5	304198500.6	286038854.3	319205600	327002316.9	317262553.1	347711728.2	348340412.2	350834897.4	379310960.6	350789558.6	389943544.5
creatinine	37254349.46	35717083.45	36632573.49	38478267.33	41172359.82	37074325.97	41084633.27	41426754.63	51663206.53	44945138.53	36742649.02	51628111.72
CTP	23118589.53	29592726.08	20291696.77	55952490.6	41591980.99	40159856.86	67541285.54	75342984.24	50061118.47	53616714.18	78804621.73	70024398.84
Cysteine_Ellmans_conjugated	57549392.8	58574814.58	57188410.69	41863589.43	45822327.27	39697617.95	24361479.53	28371203.98	28220080.62	15749226.88	12264224.72	19028389.06
cystathionine	1754507.833	1652655.624	1952975.941	4893087.544	5174713.61	5179499.963	3681726.313	3380745.918	3790248.674	3641168.747	3206057.258	3206576.011
dihydroxyacetone_phosphate	9584287.585	13391566.35	10218091.59	7942779.546	9197834.08	8686444.101	4770556.946	4929279.853	5569448.781	4817320.08	4898255.504	5358466.133
F6P_or_G1P	16734900.17	14070767.73	13210281.85	9932619.069	9680845.948	10817277.23	24042431.13	22255358.79	20950276.8	18111636.68	17596012.11	15816682.78
Fructose.1.6.bisphosphate	4375337.926	4844197.696	4188408.49	4372229.35	4243940.076	4120609.122	4451372.003	4141359.21	3929770.027	3369544.778	3662617.747	3465060.087
fumarate	234253824	205689155.6	206280729.2	174548505.1	179893272.5	172693762.7	155065940.9	141044678.8	167489937	160468706.4	153219547.2	146715237.8
GDP	11703258.05	9951619.917	10932955.4	9237507.719	8189784.939	8604913.456	11853545.45	12343850.34	11432147.1	12307805.25	12737335.27	11814408.48
Glucose.6.phosphate	9113714.305	11809277.69	10896550.66	1959130.672	2030305.044	1845351.531	1351149.069	758156.1199	1185794.081	245044.6087	710290.6123	538846.1121
glutamate	1027363289	945940694.8	1035295374	2340705989	2397560563	2511711095	2644249212	2715868822	2863946061	2969823548	2836587317	2785490450
glutamine	1641819951	1574176182	1684168459	2650362560	2585300903	2670856880	2788902258	2740275515	2894492904	3074443261	2985555723	2943171220
glycerate	43279546594	38576972203	37606325130	24284653057	25552330194	25519267310	29719752664	26178181745	26608855208	29812701237	26608784360	26895010707
Glycerol.3.Phosphate	14022963.55	15096550.47	13871719.66	8404994.211	8706777.679	8376201.775	7676619.115	6824457.467	7374149.115	5910907.847	5921713.302	5877584.874
Glycerophosphocholine	3820040302	3825841873	3854212463	3345943684	3343189808	3431028307	1440150005	1490660779	1608281378	1467238899	1566946018	1589481225
glycine	48775698.77	42926500.15	46619440.19	35890545.54	35470542.2	36963022.48	41235905.46	36579488.05	39099403.19	45297725.3	40011476.62	40355997.17
GSH_Ellmans	1118645914	1136494710	1090104727	809166698.2	826119432.1	837950751.8	653509499.1	657100342	548194916	595950966.6	607834138.3	618558046.7
GSSG	3988756.354	4131424.01	3877016.848	4266770.465	4539819.382	4027122.344	1964804.432	2246105.74	3236404.134	2186924.839	3127050.557	2020674.943
GTP	26691341.88	28676534.98	18978104.84	61103528.56	41299507.36	36650029.06	69397529.89	77776817.34	43997407.11	50717439.96	80497894.02	61537531.12
histidine	231554481.3	222976390.9	235293941.5	211386657.7	220390125.7	207602410.2	241110296.3	231925418.3	235286828.8	265414667.6	247902416.5	245255694.3
hypotaurine	3063315.35	2862655.914	3636710.899	4159432.784	4702874.401	4786775.219	5207318.098	5194960.726	5592099.7	5197668.384	5310210.929	5263592.552
IMP	2068795.461	2029243.646	2353400.269	1647063.258	1450843.772	1057212.21	2068533.998	1986164.99	1895443.471	1790041.052	1679266.081	1732595.259
isocitrate	1206745105	1289754136	1206015227	1850861407	1427861400	1420145977	930815858.6	916229983.3	1062303287	1091732558	955221329.4	976941142.4
isoleucine	380075831.9	433228238.4	448776340.6	448633617.5	459425728.7	503944115.1	430498928.1	469731090.9	480679750.3	532651766.4	448100695.8	513399686.4
lactic_acid	599383165.5	536721668.3	667720379.8	297930457.7	347401933.3	319262172.3	175024474.4	191097873.7	200074197.5	231623977.8	251140167.5	269612328
leucine	415445461.4	428318339.7	472528334.5	481784164.2	526342540.8	518367821	492297388.7	520444298	530615816.9	514678455.6	535401160.2	542042580.3
lysine	39935371	44920940.89	41841763.53	31388507.89	31076993.53	30896588.06	32897995.92	35275966.51	32486598.17	35777529.45	35243588.77	39237629.82
malate	2306582479	2097886329	1969904252	1723349164	1722855126	1717108792	1582092500	1423350423	1579249457	1640402191	1512266536	1418288586
Methionine.sulfoxide	366679228.2	358457689.9	365944968.8	392146912	431508593.6	413423361	396604469.6	389678063.4	471984856.5	439196770.9	406867346.7	459675373.4
methionine	40222740.68	45951118.26	63623446.57	59639591.69	78632911.17	88899575.71	68476792.66	78034384.45	73782005.46	77166541	85724341.91	105740348.6
methylthioadenosine	2458744.248	1963807.239	2243125.704	6607517.711	7698089.352	6881675.95	6572275.998	7868718.08	10226913.81	9197280.722	8734632.733	7063192.726
N.acetylaspartate	33797704.41	26728992.88	27319353.99	16694613.88	17561709.28	16020933.58	23175595.3	21946797.92	21390859.62	21496791.21	18830803.31	21198529.36
N.acetylglutamate	54484447.86	43179598.99	54139592.55	26214615.3	24495612.37	20738009.94	35471059.07	30998007.17	34586606.39	36182029.52	29926585.59	36100882.69
NAD.	66826229.13	66634570.81	65325168.77	109416530.3	106913257.2	106133776.6	111503814	111751192.3	115566510.1	111617208.1	118294964.3	115120774.9
NADH	7412572.193	9771540.564	9002660.594	12752906.92	13387130.46	14042818.41	14068961.22	12960613.94	5635839.967	9937741.23	11240113.56	10362859.77
NADP	1753730.315	1832019.952	1280117.062	2867138.802	2450770.629	2097543.021	2548871.692	2486531.225	4662157.882	3259719.917	3249133.314	2915639.999
NADPH	1406936.052	1833683.371	2037235.623	3486204.816	3580806.81	3755243.508	4466467.503	4340599.46	1824129.368	3103652.283	3339971.128	3631036.532
niacinamide	72812151.92	66440412.09	59689737.99	59319596.06	56137902	55631686.01	52663137.5	57369168.28	63052140.68	69851730.52	61884398.87	59753343.99
O.phosphoethanolamine	18607691.14	19030244.71	21103498.77	9082289.781	8960617.116	9235576.868	9277982.492	10348313.72	10536725.77	9503438.424	9891590.988	9130153.334
ornithine	6281053.962	6477633.097	6299185.009	4068885.318	3425893.963	4763603.525	5068229.856	6237186.659	4506186.084	6165429.395	4554385.591	4291998.016
palmitate	223307941.4	162974018.2	209720034.2	132371737.4	117712602.3	138539571.2	143395131.7	118506539.8	138262873.4	199060167	124773193.8	127786618
Pantothenate	86506573.85	79242938.16	82112828.6	187164201.8	206637716.7	195914734.3	197820648.8	185507271.8	226963958.3	201136298.7	196236775.8	196225522
phosphocholine	9132147871	9466566149	9474178685	10981898019	11182478213	10851122275	10299019419	10515986589	10167681900	10631236590	10777766673	10411456064
phosphocreatine	1927923.224	1765051.734	1402820.312	2107365.41	2648734.109	2766946.609	2099189.247	2216834.764	2064303.827	1871651.289	2009892.209	2032454.138
phosphoenolpyruvate	883006.1794	847104.9466	1196511.083	4190777.578	4938164.764	4750972.647	1958490.833	2251963.968	1864677.271	1446057.24	1462564.539	1676552.578
proline	3068961020	2649331402	3247448957	3712735839	3723269328	3826345833	3899539308	2888061947	3267331617	3705703186	3931989564	3572560188
propionylcarnitine	23779724.67	21817605.81	23373913.45	28545789.68	30542668.62	29078635.89	34515996.7	31276072.81	42025444.64	38158509.03	38480548.19	40667476.5
sarcosine	91744049.74	80812467.61	76610531.17	72208787.79	74557472.31	75008820.63	66267106.5	77555415.49	78412421.92	87880019.35	79403489.01	80719214.53
Sedoheptulose.7.phosphate	6731714.569	6384595.603	7533495.672	7236668.906	7162478.827	7101434.017	14659411.4	13694390.38	12531770.78	12459093.59	12134581.06	12063872.63
serine	191137061.5	155537725.3	169453112.1	119890093.6	119040782.8	122835157.4	145084450.6	133680822.7	138993228.3	152295088	145335550.7	135452421.2
succinate	241471464.4	215377434.3	232141287.3	140094038.5	118962181.8	122384453.2	132706617	133591530.1	162118728.3	148035111.1	143974432.5	141393198.2
taurine	311500245.4	311204365.5	319496735	355764463.9	368241528.2	368962364.5	402554039.4	400573297.1	400203475.5	426584454.7	395131577	412953105.4
threonine	56335825.89	75871033.16	91354869.41	124924177.9	93259442.05	142095248.7	116211066.9	138480533.1	179731121.3	157163189.9	141921222.2	142502131.1
trans.urocanate	277363383.5	315539059.1	304669778.9	217653502.7	180252076.9	203053695.2	211353110.6	216920792.6	194749056.2	201414163.5	215743593.8	199556896.8
tryptophan	36876133.43	38189298.56	38469856.48	41117322.57	38819501.4	47091857	44047657.52	45453488.07	46739226.99	40576434.99	46235532.52	46833607.17
tyrosine	114133411.2	127464438.8	137945373.2	126774704.6	135328543	154993571.5	131483992.9	135513320.8	139937067.5	131317919.7	138585009.2	161583086.7
UDP.GlcNAc	38781777.95	40357413.27	39083197.85	54004287.1	53350393.83	53067247.62	52783587.44	52698931.38	54069330.45	55034061.15	51789844.56	52184071.54
UDP	8871344.796	9712483.762	9439331.436	13172806.54	11545626.92	10793872.4	16757495.69	18477523.71	18182134.17	17252700.05	19501473.71	19662254.36
Ureidosuccinic_acid	3781020.625	4278859.105	3303994.105	3765365.207	3611371.15	3242831.258	3999408.218	3787784.751	4574841.242	4689493.519	4662408.124	4667106.237
UTP	60250246.37	77677968.87	55690785.4	165247925.6	123178245.3	120832578.7	208289255.7	232510509.4	158265394.9	171930360.4	237568620.3	194939807.2
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
2-aminobutyric_acid
3-phosphoglycerate
Acetyl.CoA
acetylcarnitine
ADP
alanine
Alpha.Ketoglutarate
AMP
arginine
argininosuccinate
asparagine
aspartate
asymmetric.dimethylarginine
ATP
Butyrylcarnitine
carnitine
cis.aconitate
citrate
citrulline
creatine
creatinine
CTP
Cysteine_Ellmans_conjugated
cystathionine
dihydroxyacetone_phosphate
F6P_or_G1P
Fructose.1.6.bisphosphate
fumarate
GDP
Glucose.6.phosphate
glutamate
glutamine
glycerate
Glycerol.3.Phosphate
Glycerophosphocholine
glycine
GSH_Ellmans
GSSG
GTP
histidine
hypotaurine
IMP
isocitrate
isoleucine
lactic_acid
leucine
lysine
malate
Methionine.sulfoxide
methionine
methylthioadenosine
N.acetylaspartate
N.acetylglutamate
NAD.
NADH
NADP
NADPH
niacinamide
O.phosphoethanolamine
ornithine
palmitate
Pantothenate
phosphocholine
phosphocreatine
phosphoenolpyruvate
proline
propionylcarnitine
sarcosine
Sedoheptulose.7.phosphate
serine
succinate
taurine
threonine
trans.urocanate
tryptophan
tyrosine
UDP.GlcNAc
UDP
Ureidosuccinic_acid
UTP
METABOLITES_END
#END