#METABOLOMICS WORKBENCH alanyl_20241007_115037 DATATRACK_ID:5265 STUDY_ID:ST003819 ANALYSIS_ID:AN007387 PROJECT_ID:PR002390
VERSION             	1
CREATED_ON             	December 3, 2025, 6:19 am
#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 SLC31A1 knockout cells compared to intergenic
ST:STUDY_TITLE                   	sgRNA containing cells
ST:STUDY_SUMMARY                 	To investigate the effects of SLC31A1 knockout on steady state metabolite
ST:STUDY_SUMMARY                 	levels, SEM (acute lymphoblastic leukaemia cell line) cells containing either a
ST:STUDY_SUMMARY                 	SLC31A1-targeting sgRNA (two different guides, guide #2 or guide #4) or a
ST:STUDY_SUMMARY                 	intergenic -targetting sgRNA (i.e. a control sgRNA, "Intergenic1") and Cas9 were
ST:STUDY_SUMMARY                 	FACS-purified (Fluorescence Activated Cell Sorting) based on SLC31A1 expression.
ST:STUDY_SUMMARY                 	Cells were then cultured in RPMI-1640 containing 10% FBS and penicillin and
ST:STUDY_SUMMARY                 	streptomycin in 6-well plates, and then collected for metabolomics. SLC31A1
ST:STUDY_SUMMARY                 	knockout cells were found to exhibit profound electron transport chain
ST:STUDY_SUMMARY                 	inhibition, resulting in accumulation of carbamoyl aspartic acid/ureidosuccinic
ST:STUDY_SUMMARY                 	acid, and depletion of aspartate, as well as broad changes in nucleotide mono,
ST:STUDY_SUMMARY                 	di and triphosphate 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           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1058	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1058.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1059	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1059.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1060	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1060.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1061	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1061.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1062	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1062.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1063	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1063.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1064	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1064.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1065	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1065.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1066	Sample source:SEM leukemia cells | gRNA:Intergenic1	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1066.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1067	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1067.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1068	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1068.raw
SUBJECT_SAMPLE_FACTORS           	-	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1069	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4	RAW_FILE_NAME(Raw file name)=20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1069.raw
#COLLECTION
CO:COLLECTION_SUMMARY            	One million cells from culture were collected via centrifugation for 20 seconds
CO:COLLECTION_SUMMARY            	at 18,000xG at 4C, washed once with ice-cold 0.9% NaCl, and collected via
CO:COLLECTION_SUMMARY            	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 harboring control or SLC31A1-targetting guideRNAs were treated with
TR:TREATMENT_SUMMARY             	doxycycline at 2ug/mL for 3 days, then FACS purified for SLC31A1-hi (in the case
TR:TREATMENT_SUMMARY             	of control guideRNA) or SLC31A1-lo (in the case of the SLC31A1-targetting
TR:TREATMENT_SUMMARY             	guideRNA). After sorting, cells were expanded in vitro.
#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.
#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 acetonitrile; buffer B was 20 mM ammonium carbonate,
CH:CHROMATOGRAPHY_SUMMARY        	0.1% ammonium hydroxide in water; resulting pH is around 9 without pH
CH:CHROMATOGRAPHY_SUMMARY        	adjustment. Gradient conditions used were: 0-20 min: linear gradient from 20% to
CH:CHROMATOGRAPHY_SUMMARY        	80% B; 20-24 min: hold at 80% B; 24-24.1 min: from 80% to 20% B; 24.1-32 min:
CH:CHROMATOGRAPHY_SUMMARY        	hold at 20% B at 0.150 mL/min flow rate. The column oven and autosampler tray
CH:CHROMATOGRAPHY_SUMMARY        	were held at 25 °C and 4 °C, respectively.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um)
CH:SOLVENT_A                     	100% acetonitrile
CH:SOLVENT_B                     	100% water; 20mM ammonium carbonate; 0.1% ammonium hydroxide
CH:FLOW_GRADIENT                 	Gradient conditions were as follows: 0-20min: linear gradient from 20 to 80% B;
CH:FLOW_GRADIENT                 	20–20.5 min: from 80 to 20% B; 20.5–28 min: hold at 20% B.
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, San Jose, CA, USA) and was performed with polarity switching. Scans
MS:MS_COMMENTS                   	used were in the positive and negative ionization mode in a range of m/z =
MS:MS_COMMENTS                   	70–1000, with the resolution set at 70,000, the AGC target at 1 × 10^6, and
MS:MS_COMMENTS                   	the maximum injection time (Max IT) at 40 msec.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	Normalized peak area
MS_METABOLITE_DATA_START
Samples	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1058	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1059	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1060	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1061	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1062	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1063	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1064	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1065	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1066	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1067	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1068	20240119_QE2_HILIC_SLC31A1ko_metabolomics_AYW1069
Factors	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#2	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:Intergenic1	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4	Sample source:SEM leukemia cells | gRNA:sgSLC31A1-#4
2-hydroxyglutarate	244345767.3	276318517	251239261	438226255.2	357492303.4	497662360.3	103842577.1	95795134.18	103453000.5	164466342.1	145285493.6	138575191.2
3-methyl-2-oxopentanoic_acid	22953647.19	28738697.75	24703528.6	16968819.04	22190288.82	22140629.58	20029999.16	17592123.65	18578074.43	17916789.36	15261150.27	16129899.54
3-phosphoglycerate	4496454.81	4086827.586	3907391.571	2624023.906	2735267.188	2419483.435	2395646.595	2502227.646	1976981.975	1237933.758	1413743.045	2277129.169
Acetyl.CoA	5215725.189	5323410.927	5915549.343	4294906.604	4296027.399	4399560.704	7199819.686	7917164.894	7157444.292	6197734.087	5872644.197	5876696.1
acetylcarnitine	174869885.1	168901447.4	180032368.8	176959308.3	182590305.9	182547053.4	244912117	245571460.5	250617434.8	223345532.6	222934150	213065777.9
ADP	33512477.38	28193523.9	30275302.46	53352078.43	52908048.49	51362715.47	37403885.02	36694420.06	33981743.96	47523446	53866961.83	50515525.86
alanine	127867481.7	114006369.1	131230367.1	160809317.1	152105514.8	154636912.7	263279763.9	240822233	261093910.4	282037109.4	277880690.4	261120022.1
Alpha.Ketoglutarate	110522830.7	109621815.2	120994106.1	79239227.16	91773893.97	94089295.03	84398030.35	71605051.46	82233981.46	74092505.3	74689301.7	71497010.68
AMP	4652536.356	3992666.925	4680120.291	10637580.8	11152683.92	10752820.4	4110125.784	4547176.205	4368609.437	9053548.204	10315937.86	8599800.164
arginine	3742568751	3930654986	3797829742	3440753603	3171473862	3192597483	3406240855	3300689618	3178908715	2885292982	2691552888	2837303117
argininosuccinate	36389568.92	34782908.97	38047012.42	33778634.18	35810952.25	35346450.55	10242985.9	9747344.493	11010435.63	12825606.21	12111702.65	12765690.37
asparagine	203559525.9	186183013.5	200948628.1	153780120	149788833.3	150340235.1	214293887.4	214158794.4	224588087.9	170009683.4	164058929.7	177089376.5
aspartate	28397730.28	27166044.46	27647251.74	16513873.4	16495295.06	15900888.47	34414767.35	34590527.9	36506788.5	17000256.59	15039835.64	16799183.63
asymmetric.dimethylarginine	25825181.13	30611166.29	28155919.42	21368570.89	21304188.38	21936332.83	25706351.49	25612774.62	24923145.78	20947102	20596016.08	20023620.1
ATP	244565778.8	178100050	182030251.4	201106204.7	186725817.6	183349584.4	249943467.5	260610174.9	247382907.5	200781235	233571059.8	218391983.9
beta.alanine	72068388.12	56772875.87	76293619.76	75393603.14	70186831.28	70048032.28	190629191.2	171863785.4	181678943.9	220841293.4	202012182.3	196953371
betaine	1544974896	1507713573	1580235407	1116765600	1046867307	1094907478	2025283161	1999341944	2047115150	1911909222	1810759038	1753424381
Butyrylcarnitine	158567222.4	143459530.7	160672479.2	150767333.5	154533829.1	158154665.1	206954513.2	201779241.8	210567408.8	197146344.8	189771219.9	195615295.9
C5.Carnitines	8146649.233	7926497.897	8837265.872	20441880.04	19629169.69	19538282.64	9918598.833	9735102.452	9381027.162	30084665.24	28735668.24	28713955.25
carnitine	278902248.6	265379917.9	288793876	292547407.2	285243260.6	282180114	364513219.6	350124675.4	365391433	394093934.9	393366375.5	382092715.4
carnosine	3203379.256	5314287.97	3686706.663	4040917.524	3854468.007	3661264.896	2067595.562	2117377.735	1882935.107	1946569.736	1685184.211	1908523.223
CDP	1242376.564	1056820.504	636449.2624	1920880.581	2189007.933	1871930.898	757509.6608	1041613.871	967671.9172	1042768.601	1300094.231	1365735.96
choline	20022739.44	15884920.94	20381264.59	13914686.95	14909917.74	16740072.76	29411500.29	32226684.61	30106484.56	21293035.7	18714480.65	20481407.18
cis.aconitate	25934855.34	24558564.31	26494782.35	10581382.61	13140908.86	11264908.95	25911561.2	21589501.7	26909625.1	16831545.5	16140329.86	15151479.91
citrate	966815937.8	868903306	923795697.8	518640851.9	544810154.4	517945617	579010986.3	528995817.9	547300576.2	436097655.3	416103996.8	367094122.9
citrulline	22169469.22	22234224.31	24276594.53	21726436.32	20821801.18	21932549.24	17433190.15	18887598.04	21817697.03	18743849.1	19526350.91	23625135.43
creatine	1036078543	1092095824	1087032306	982314851.8	1020286267	994101716.8	1039034733	992890252	977307524.2	1016157514	996020667.9	955283708.8
creatinine	88848593.17	100173650.6	92275365.8	85070499.7	85658552.15	86259299.09	97168504.78	95541808.01	91544257.97	87544256.76	88197382.93	85621327.5
CTP	22692362.32	15296294.2	15394011.38	11676193.99	10834151.14	10599301.08	23682843.05	24717657.96	22760162.26	16140752.5	19118969.68	17566225.64
cystathionine	6987970.06	6824885.032	7587294.468	4253995.673	4190424.95	4006923.663	2939249.593	2883936.679	3378730.432	2396848.271	2102246.347	2057399.204
cystine	3823294.705	10851739.62	4166140.127	6856369.761	6217164.042	5836367.415	1327145.018	2651094.636	2150589.397	2063631.461	1203174.574	767736.8405
dATP	1048694.114	922037.4144	943851.3151	890402.4048	864310.1124	731838.91	916687.0131	1270748.114	843866.9532	255597.7132	442478.044	101634.4209
dihydroxyacetone_phosphate	593146.4725	555143.5093	479493.2714	360373.2246	785636.1319	474137.0253	228542.6307	232682.3927	332398.5626	54562.75147	351896.848	80080.23511
F6P_or_G1P	1809719.182	1456374.06	1468107.988	1625585.15	1620248.77	1790881.473	1051360.32	943030.2582	848038.1796	1936973.989	1802076.168	1843547.871
FAD	1364890.663	1296805.031	1293311.125	1216608.805	1213632.478	1540948.113	1182598.655	1314952.827	823422.0858	778784.7505	1262034.127	1084640.916
fumarate	50932568.39	43118256.88	49547932.75	53061988.21	54310533.88	53101238.66	69619289.54	71886161.16	69977972.67	76474040.45	72904552.76	70071436.6
GDP	3131169.325	2336871.775	2383544.184	5445224.763	5796564.039	5244859.243	4065594.358	4241289.192	3439842.664	5807101.37	7004861.187	6070736.303
glutamate	1149705167	1069237991	1168915948	921520451.5	897306348	901576280.5	1035140717	1033749027	1055516397	882405041.9	839932521.2	876299999.6
glutamine	5354801739	5048194056	5504571880	4372788985	4309311725	4156561171	4536172166	4372478157	4735466768	3731060321	3485852865	3721506624
glycerate	1124212713	996749031.8	1161729394	1067202530	991490265.3	990080235.1	17552402569	15561967535	17818538047	18658439513	16998377454	16554067462
Glycerol.3.Phosphate	13028711.67	12795729.73	12523522.68	34079229.63	35481030.26	35146644.92	8552715.794	7705103.475	7559512.736	37312286.32	36983500.77	36941339.82
Glycerophosphocholine	4445673920	4040648243	4767065383	3694436199	3665627730	3852376104	7575778015	7617610618	7532544676	5319805738	4992690244	5161597720
glycine	13887900.93	10463748.52	12913221.31	11214549.33	12691540.55	11381916.31	57832533.85	52158684.43	57575655.35	57842323.66	56368227.65	53209618.81
GMP	321655.1292	121677.5873	425580.6047	1348425.85	1534337.929	1709273.577	463766.0331	546928.791	66794.6168	902500.5589	1371235.75	1224358.696
GSH_Ellmans	301722108.5	268021151.9	298067764.8	281000287.7	263412617.9	259717634.1	242265603	249412643.9	250805163.9	231366038.3	230600806.6	228436118.7
GSSG	5090995.069	4057739.849	4559234.794	4005134.334	5401994.542	4550593.606	3205403.924	1918106.816	1594256.113	1469286.414	1611993.44	1855052.032
GTP	20806319.53	14716641.2	14222564.78	17479006.7	17103044.51	16813442.66	30266483.51	30053810.43	24893550.28	25680502.44	31882092.76	28309349.25
histidine	708112726.4	751528176.3	738810903.3	620606987.9	655681880.9	631920297.4	352669935	310170654.7	338576349.7	284881947.3	297792952.8	276975434.7
homocitrulline	2527851.469	2461449.23	2360450.642	3816365.259	1623421.108	2040051.663	2227198.552	2434685.729	2974011.483	3741673.934	2536125.661	4743099.222
hypotaurine	3590662.686	4224990.576	3822966.618	2900524.173	8709471.749	5252540.994	9757335.607	9599004.241	11386146.09	12196246.4	12933324.91	9445046.831
IMP	783016.064	631243.3303	676936.4191	1334656.681	1349903.172	1373597.192	184109.5246	97179.34055	55376.04896	1596892.888	1835725.191	1229830.217
isoleucine	883861950.5	891474741.9	891197831.5	801042809.6	802749665.3	777915264.8	734223110.2	706334653.8	769442043.1	676582527.6	680470559.8	666484821.9
Itaconic.Acid	NA	NA	NA	NA	NA	NA	19023627.02	20015268.93	16853831.23	18486049.75	17952909.57	18521241.78
lactic_acid	400208393.1	416353213.3	400766887.1	577500774.8	644646357.4	647730960.7	321629328.3	329454570.3	291460237.6	604679422.5	509526687.4	483718235.9
leucine	803370986	809829896.3	804803780.4	723128715.5	743534660	717545807	694989567.4	678802843.8	729404736.5	630790696	620046658.5	622818646.6
lysine	69556279.42	72220011.73	66449214.41	76963089.54	67073317.4	66529790.35	28168092.67	27848742.24	26087277.8	24660400.5	22195350.66	23358556.69
malate	630447309.6	564920435.8	629049646.8	648706342.3	654093641	638801984.8	913102693.2	847088209.8	927536657.9	993952365.9	926165744.2	945842378.5
Methionine.sulfoxide	119217211.8	122491548.8	126468472.9	94255202.51	103375621.5	104714074.4	456946754.8	445800118.5	467193266.3	386712667.6	385544124.8	381175297.9
methionine	264430085.3	280197372	272581123	238152471.1	270552320.9	245675729.3	113843515.7	106276138.8	115416306.1	90302380.12	94214291.71	89132830.77
methylthioadenosine	13379749.13	10910119.83	12893534.21	9551743.838	8635258.688	9569419.136	12449580.84	11540374.41	9506484.357	10087784.38	8760983.798	10686824.14
N.acetylaspartate	5509388.018	4886740.923	4807250.483	4286926.521	2938715.171	3568554.339	34377972.08	32127231.07	33571627.16	33136758.24	33940321.96	33780527.23
N.acetylglutamate	33533538.35	31375679.73	35536564.27	32987194.28	34788573.03	32537359.38	101955447	93766320.09	110440967.9	120168789.4	111047230.1	95301021.74
NAD.	102443612.1	92648835.74	106051471.9	87629218.04	92775963.29	92602685.43	119945222.1	121254571.6	117490065.6	108877537.8	106166917.8	103993967.9
NADH	20727113.09	18085739.56	20293811.55	26513688.12	28124154.41	26284210.53	16217029.58	15342225.18	13910775.77	24111056.88	23233284.03	22967626.51
NADP	1427149.81	1559124.434	1497434.523	1532165.46	1604581.234	1656954.234	1377795.921	1236854.431	1578599.369	1519436.055	1653744.678	1656013.083
NADPH	4291090.623	3820728.061	3941202.377	3162392.747	3393517.484	3251532.205	5408745.599	4941512.829	4796894.798	4004723.533	3947000.205	3747653.763
niacinamide	91667915.27	106047448.6	110587575.5	116357339.8	109669942.1	115372064.9	149301920.9	163715505.3	153146946.1	193969917.7	166886374.1	161653569.8
ornithine	54817760.01	50286515.32	42282646.4	42961795.79	36019821.52	35704771.69	8647252.199	8212535.612	8035859.855	6865827.234	7042794.905	6915987.745
orotate	NA	130105.362	NA	NA	NA	NA	45934821.1	38795939.18	44015111.95	44652779.75	41683601.97	41996670.29
palmitate	NA	373187534.9	343720684.8	349857790.4	NA	258294201.7	NA	196823557.1	303104794.1	NA	NA	278224821.7
phosphocholine	33402716364	30932827120	33384148116	26608949476	27056960069	26169174180	36046356966	33943505294	35262172914	32523093500	30357212345	30625354095
phosphoenolpyruvate	1899268.722	2018500.615	1773269.855	830363.3247	561339.7061	616361.223	392755.4529	708764.3937	560834.4783	220587.8641	274109.9315	522165.4915
proline	7524512128	6957559129	7629105725	7326855699	7141148441	7151698079	6953248555	6615489959	7208322929	8263799523	8099668232	8120835584
propionylcarnitine	7537496.775	8630658.945	8578783.85	3317282.89	3351585.995	3353020.316	15232096.14	15335404.74	14757287.25	5901927.907	5726612.472	5819817.596
Pyridoxal	6099090.133	5982617.673	8314981.551	4548058.597	2788340.575	2576843.074	77576246.76	80655733.5	58069052.01	127741677.9	59807582.27	117008849.4
Pyridoxine	142268304.9	153722626.2	150941596.5	134689454.5	139094057	136958422.5	138610799.3	142186596.9	147289694.2	127229438.9	150817709.3	140028897.7
S.Adenosyl.L.Homocysteine	19506810.25	18427810.51	20082413.73	16539159.46	17521238.39	17251478.23	6898204.248	6619300.096	5866553.141	6268374.571	6192012.565	6567268.144
S.Adenosyl.L.methionine	303802971.9	287313486	317531314	264832747.8	265132048.1	257542631.7	179668678.2	169508381.8	165457398.4	163825782.4	171647608.6	162236209.7
saccharopine	2522456.812	2205528.736	2359900.672	1071019.123	1231886.423	1169516.265	1548767.922	1489497.475	1642292.765	1223597.914	1197302.16	732128.38
Sedoheptulose.7.phosphate	1361592.954	1089462.804	989208.8921	2667292.326	2356989.293	2764547.02	319667.7103	764953.7514	383846.079	1083599.083	1084904.788	1384041.617
serine	26306958.78	26504720.93	28128159.59	31714300.58	29912271.29	30230949.54	260672444.6	241402892.6	254284343.3	266159217.2	264119972.4	244287639.2
succinate	27835392.51	31808103.46	30575704.42	87026461.66	86817979.83	90343041.38	87341175.4	84648188.45	90233283	116762701.1	114116415	107668533.9
succinyl.CoA	1355963.91	1043499.854	1576684.817	485991.8885	276395.5755	108120.7835	1153479.8	1719565.727	1632974.754	487771.5263	555404.6554	434770.3957
taurine	191274458.4	178281212.5	194476962.9	170692410.2	172492639.8	169862522.4	169572399.2	170101507.2	172348375.5	205388201.7	197723619.5	197931953
threonine	148937643.1	146468256.6	140721391.3	119147956.9	127115408.4	145028692.1	244885710	160620828.9	221417403.5	197892077.9	182210219.5	196260626.1
trans.urocanate	387365245.6	223267275.2	273241057.2	234400046	182755963.1	218176568.4	1024641638	919493320.7	1091117466	1081479989	945652476.6	975103797.4
tryptophan	79937726.45	85699648.55	84547809.46	74636319.66	82135187.24	76157804.4	36844954.08	36064390.49	37555000.11	32078369.2	33183156.32	31824282.51
tyrosine	233872475.8	257084193	253330992.1	213366662.8	239591540.5	231252276.9	177379069	162052749	182323626.1	140812514.1	149395735.4	129725487.1
UDP.GlcNAc	34140200.29	31808161.3	36465271.26	18428420.32	20095044.56	19760907.69	29562911.62	28162418.45	28415569.33	18246374.88	18765227.83	17068314.87
UDP	3960049.995	3068960.381	3089214.263	6279286.767	7036430.423	6530544.052	4020621.115	3233500.582	4225605.354	5212053.176	5658069.755	5706651.554
UMP	525848.6363	693156.1273	744363.007	1718567.796	1818207.279	1985252.578	625834.4869	714826.7667	373014.5196	1293489.767	1481631.265	1739073.226
Ureidosuccinic_acid	14212514.1	13804598.9	14999277.21	116985436.6	117696405.4	118390313.3	9714946.06	10878325.69	10542014.92	32398842.08	31011018.77	33833535.2
UTP	62490314.96	49186816.8	48267421.76	39191501.57	35657748.55	35670244.41	67721198.64	69121104.34	64366927.17	47886869.87	56537788.53	53693634.08
valine	237984667.9	254096347.9	215612041.4	249341889.3	249548984.6	253731935.9	202784597.8	198966870.9	223679629.7	148201864	193363288.9	135503836.7
xanthine	1998466.976	2171589.766	1875734.606	1816178.705	2156355.388	1847971.178	1496769.166	1060247.36	1023645.734	1150753.334	1248683.526	1251650.636
dihydroorotate	3181676.626	2389125.011	2725737.513	24472369.69	24092092.46	24859054.58	1757846.883	2351331.7	2060346.184	6211898.224	5447321.846	6344343.723
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name
2-hydroxyglutarate
3-methyl-2-oxopentanoic_acid
3-phosphoglycerate
Acetyl.CoA
acetylcarnitine
ADP
alanine
Alpha.Ketoglutarate
AMP
arginine
argininosuccinate
asparagine
aspartate
asymmetric.dimethylarginine
ATP
beta.alanine
betaine
Butyrylcarnitine
C5.Carnitines
carnitine
carnosine
CDP
choline
cis.aconitate
citrate
citrulline
creatine
creatinine
CTP
cystathionine
cystine
dATP
dihydroxyacetone_phosphate
F6P_or_G1P
FAD
fumarate
GDP
glutamate
glutamine
glycerate
Glycerol.3.Phosphate
Glycerophosphocholine
glycine
GMP
GSH_Ellmans
GSSG
GTP
histidine
homocitrulline
hypotaurine
IMP
isoleucine
Itaconic.Acid
lactic_acid
leucine
lysine
malate
Methionine.sulfoxide
methionine
methylthioadenosine
N.acetylaspartate
N.acetylglutamate
NAD.
NADH
NADP
NADPH
niacinamide
ornithine
orotate
palmitate
phosphocholine
phosphoenolpyruvate
proline
propionylcarnitine
Pyridoxal
Pyridoxine
S.Adenosyl.L.Homocysteine
S.Adenosyl.L.methionine
saccharopine
Sedoheptulose.7.phosphate
serine
succinate
succinyl.CoA
taurine
threonine
trans.urocanate
tryptophan
tyrosine
UDP.GlcNAc
UDP
UMP
Ureidosuccinic_acid
UTP
valine
xanthine
dihydroorotate
METABOLITES_END
#END