#METABOLOMICS WORKBENCH JasonMiska_20240614_100943 DATATRACK_ID:4922 STUDY_ID:ST003262 ANALYSIS_ID:AN005346 PROJECT_ID:PR002024 VERSION 1 CREATED_ON June 17, 2024, 9:54 am #PROJECT PR:PROJECT_TITLE A Covalent Creatine Kinase Inhibitor Ablates Glioblastoma Migration and PR:PROJECT_TITLE Sensitizes Tumors to Oxidative Stress PR:PROJECT_TYPE LC-MS/MS PR:PROJECT_SUMMARY Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, PR:PROJECT_SUMMARY diffuse infiltration, and near-uniform lethality. The underlying mechanisms are PR:PROJECT_SUMMARY unknown, and no treatment has been curative. Using a recently developed kinase PR:PROJECT_SUMMARY inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. PR:PROJECT_SUMMARY While CKi minimally impacted GBM cell proliferation and viability, it PR:PROJECT_SUMMARY significantly affected migration. In established GBM cell lines and PR:PROJECT_SUMMARY patient-derived xenografts, CKi ablated both the migration and invasion of GBM PR:PROJECT_SUMMARY cells. CKi also hindered radiation-induced migration. RNA-seq revealed a PR:PROJECT_SUMMARY decrease in invasion-related genes, with an unexpected increase in glutathione PR:PROJECT_SUMMARY metabolism and ferroptosis protection genes post-CKi treatment. The effects of PR:PROJECT_SUMMARY CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 PR:PROJECT_SUMMARY metabolite tracing indicated heightened glutathione biosynthesis post-CKi PR:PROJECT_SUMMARY treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis PR:PROJECT_SUMMARY activation abrogated cell survival. Our data demonstrated that CKi perturbs PR:PROJECT_SUMMARY promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase PR:PROJECT_SUMMARY axis as a druggable target for GBM treatment. PR:INSTITUTE Northwestern University, Feinberg School of Medicine PR:DEPARTMENT Neurological Surgery PR:LABORATORY Jason Miska PR:LAST_NAME Miska PR:FIRST_NAME Jason PR:ADDRESS 676 N St. Clair PR:EMAIL jason.miska@northwestern.edu PR:PHONE 8478678201 #STUDY ST:STUDY_TITLE A Covalent Creatine Kinase Inhibitor Ablates Glioblastoma Migration and ST:STUDY_TITLE Sensitizes Tumors to Oxidative Stress. ST:STUDY_TYPE Cki treatment on glioblastoma ST:STUDY_SUMMARY Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, ST:STUDY_SUMMARY diffuse infiltration, and near-uniform lethality. The underlying mechanisms are ST:STUDY_SUMMARY unknown, and no treatment has been curative. Using a recently developed kinase ST:STUDY_SUMMARY inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. ST:STUDY_SUMMARY While CKi minimally impacted GBM cell proliferation and viability, it ST:STUDY_SUMMARY significantly affected migration. In established GBM cell lines and ST:STUDY_SUMMARY patient-derived xenografts, CKi ablated both the migration and invasion of GBM ST:STUDY_SUMMARY cells. CKi also hindered radiation-induced migration. RNA-seq revealed a ST:STUDY_SUMMARY decrease in invasion-related genes, with an unexpected increase in glutathione ST:STUDY_SUMMARY metabolism and ferroptosis protection genes post-CKi treatment. The effects of ST:STUDY_SUMMARY CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 ST:STUDY_SUMMARY metabolite tracing indicated heightened glutathione biosynthesis post-CKi ST:STUDY_SUMMARY treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis ST:STUDY_SUMMARY activation abrogated cell survival. Our data demonstrated that CKi perturbs ST:STUDY_SUMMARY promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase ST:STUDY_SUMMARY axis as a druggable target for GBM treatment. ST:INSTITUTE Northwestern University, Feinberg School of Medicine ST:DEPARTMENT Neurological Surgery ST:LABORATORY Jason Miska ST:LAST_NAME Miska ST:FIRST_NAME Jason ST:ADDRESS 676 N St. Clair ST:EMAIL jason.miska@northwestern.edu ST:PHONE 8478678201 ST:NUM_GROUPS 2 ST:TOTAL_SUBJECTS 12 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - Les-Jas-20230207-01 Sample source:Brain | Treatment:CTL RAW_FILE_NAME=Les-Jas-20230207-01 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-02 Sample source:Brain | Treatment:CTL RAW_FILE_NAME=Les-Jas-20230207-02 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-03 Sample source:Brain | Treatment:CTL RAW_FILE_NAME=Les-Jas-20230207-03 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-04 Sample source:Brain | Treatment:1hr RAW_FILE_NAME=Les-Jas-20230207-04 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-05 Sample source:Brain | Treatment:1hr RAW_FILE_NAME=Les-Jas-20230207-05 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-06 Sample source:Brain | Treatment:1hr RAW_FILE_NAME=Les-Jas-20230207-06 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-07 Sample source:Brain | Treatment:4hrs RAW_FILE_NAME=Les-Jas-20230207-07 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-08 Sample source:Brain | Treatment:4hrs RAW_FILE_NAME=Les-Jas-20230207-08 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-09 Sample source:Brain | Treatment:4hrs RAW_FILE_NAME=Les-Jas-20230207-09 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-10 Sample source:Brain | Treatment:8hrs RAW_FILE_NAME=Les-Jas-20230207-10 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-11 Sample source:Brain | Treatment:8hrs RAW_FILE_NAME=Les-Jas-20230207-11 SUBJECT_SAMPLE_FACTORS - Les-Jas-20230207-12 Sample source:Brain | Treatment:8hrs RAW_FILE_NAME=Les-Jas-20230207-12 #COLLECTION CO:COLLECTION_SUMMARY GBM cells were cultured under the conditions indicated in the manuscript, then CO:COLLECTION_SUMMARY metabolites were collected using 80% methanol extraction. CO:SAMPLE_TYPE Brain #TREATMENT TR:TREATMENT_SUMMARY metabolites isolated from Cki treated cells. Samples 1-3 are control treatment, TR:TREATMENT_SUMMARY Sample 4-6 are 1 hour CKi treatment, Samples 7-9 are 4 hours CKi treatmet, and TR:TREATMENT_SUMMARY 10-12 are 8 hours CKi treatment. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Isolated samples were dried using a SpeedVac. Acetonitrile (50%) was added to SP:SAMPLEPREP_SUMMARY the tube for reconstitution following overtaxing for 30s. Sample solution was SP:SAMPLEPREP_SUMMARY then centrifuged for 15min at 20,000g and 4°C. Supernatant was collected for SP:SAMPLEPREP_SUMMARY LC-MS analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Samples were analyzed by high-performance LC (HPLC) and high-resolution MS and CH:CHROMATOGRAPHY_SUMMARY MS/MS (HPLC-MS/MS). The system consists of Thermo Q Exactive with an CH:CHROMATOGRAPHY_SUMMARY electrospray source and an UltiMate3000 (Thermo Fisher Scientific) series HPLC CH:CHROMATOGRAPHY_SUMMARY consisting of a binary pump, degasser, and autosampler outfitted with an XBridge CH:CHROMATOGRAPHY_SUMMARY Amide column (Waters; dimensions of 4.6 mm by 100 mm and a 3.5-μm particle CH:CHROMATOGRAPHY_SUMMARY size). The mobile phase A contained 95% water/5% acetonitrile (v/v), 20 mM CH:CHROMATOGRAPHY_SUMMARY ammonium hydroxide, and 20 mM ammonium acetate (pH 9.0); phase B was 100% CH:CHROMATOGRAPHY_SUMMARY acetonitrile. The gradient was performed as follows: 0 min, 15% A; 2.5 min, 30% CH:CHROMATOGRAPHY_SUMMARY A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A CH:CHROMATOGRAPHY_SUMMARY with a flow rate of 400 µl/min. CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Water's Xbridge amide (100 x 3mm, 3.5 um) CH:SOLVENT_A 95% water/5% acetonitrile; 20 mM ammonium hydroxide; 20 mM ammonium acetate (pH CH:SOLVENT_A 9.0) CH:SOLVENT_B 100% acetonitrile CH:FLOW_GRADIENT 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% CH:FLOW_GRADIENT A; and 18 to 25 min, 15% A CH:FLOW_RATE 400 μl/min CH:COLUMN_TEMPERATURE - #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Plus Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE UNSPECIFIED MS:MS_COMMENTS The capillary of the electrospray ionization source was set to 275°C, with MS:MS_COMMENTS sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units, and the MS:MS_COMMENTS spray voltage at 4.0 kV. In positive/negative polarity switching mode, a MS:MS_COMMENTS mass/charge ratio (m/z) scan range from 70 to 850 was chosen and MS1 data were MS:MS_COMMENTS collected at a resolution of 70,000. The automatic gain control target was set MS:MS_COMMENTS at 1 × 106, and the maximum injection time was 200 ms. The top five precursor MS:MS_COMMENTS ions were subsequently fragmented, in a data-dependent manner, using the MS:MS_COMMENTS higher-energy collisional dissociation cell set to 30% normalized collision MS:MS_COMMENTS energy in MS2 at a resolution power of 17,500. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Peak area MS_METABOLITE_DATA_START Samples Les-Jas-20230207-01 Les-Jas-20230207-02 Les-Jas-20230207-03 Les-Jas-20230207-04 Les-Jas-20230207-05 Les-Jas-20230207-06 Les-Jas-20230207-07 Les-Jas-20230207-08 Les-Jas-20230207-09 Les-Jas-20230207-10 Les-Jas-20230207-11 Les-Jas-20230207-12 Factors Sample source:Brain | Treatment:CTL Sample source:Brain | Treatment:CTL Sample source:Brain | Treatment:CTL Sample source:Brain | Treatment:1hr Sample source:Brain | Treatment:1hr Sample source:Brain | Treatment:1hr Sample source:Brain | Treatment:4hrs Sample source:Brain | Treatment:4hrs Sample source:Brain | Treatment:4hrs Sample source:Brain | Treatment:8hrs Sample source:Brain | Treatment:8hrs Sample source:Brain | Treatment:8hrs 1-methylnicotinamide+ 7799753.722 9988125.805 101245686 5881691.603 11988434.3 10567108.93 9463413.658 12827150.42 12740823.58 6582861.107 8340249.952 9998279.773 1-phenylethylamine 26120857.13 41558361.92 20354189.53 24185851.29 18321220.64 21350833.69 23600905.79 15889503.15 21276939 14912257.24 19721635.82 11258938.66 2- / 3-phosphoglycerate 84949.89334 12877.84771 55689.24041 93153.52321 208778.4263 94520.66309 166466.4498 110833.5754 195366.2652 30983.77452 136049.0119 204319.8309 2,3-bisphospho-D-glycerate 19056.06926 29405.57087 42057.0934 13487.16913 38106.73347 18097.26655 49880.91058 31672.6562 197296.9951 0 0 23575.15544 2-/ 3- hydroxybutanoic acid 1990688.36 2399851.501 3661050.253 2572966.962 2196157.852 1889991.919 1516528.457 4792085.654 1628263.441 857144.0701 988712.9176 1278688.933 2-aminobutyric / 2-/3-aminoisobutyric acid 10855348.63 10312868.05 38023857.95 5982741.669 19529378.12 23090850.66 9095741.972 10998188.66 10740091.61 9968958.188 12001992.4 14270815.84 2-HG 699900.6745 421520.1022 2128966.604 2171668.009 1436197.529 1207052.651 1404211.686 516098.2545 632259.7387 1729399.521 2668943.245 1061870.248 2-ketovaleric acid 2923653.594 4804277.216 4131086.107 0 4448561.063 0 0 0 4004488.449 2681236.576 0 1434835.284 3- / 4-hydroxyphenylacetic acid 172996.7616 410045.9231 525886.8666 335619.2159 437487.7642 115848.6473 77095.52924 194161.7969 189288.8752 438836.5616 211274.9726 299449.8063 3- / 7-methylguanine 74899.26218 186620.5665 137118.5099 121951.4192 89176.78877 73211.48587 72589.93965 84399.15644 111363.3631 82452.95079 69990.45495 100718.0186 3-phospho-serine 27434.84259 15888.37665 46514.42233 0 36430.27987 76169.75074 0 10129.46044 67075.80272 0 0 0 4-hydroxybutyric acid 63462349.93 74428859.53 63564903.35 53276796.14 73926349.18 57914119.82 72033043.14 62581624.05 64401029.05 63377539.29 70312065.03 61829598.63 4-hydroxyproline / 5-aminolevulinate 1267747.265 2299814.746 2468877.447 3814865.906 1436432.107 2631991.738 3765681.451 4991645.363 3533753.123 4620077.039 2766671.365 1737941.529 5-HIAA 16582.2178 23005.90963 0 16333.24929 0 15231.30022 25860.20823 49959.56857 55075.57132 66430.49582 32238.58004 0 5-methylcytosine 427256.0514 510192.1725 278309.8988 497999.3672 356186.3373 261762.6216 416026.6246 230263.1075 407365.4994 418693.816 197013.4013 351058.027 5'-methylthioadenosine 8511086.144 9924309.628 11842653.11 14058280.93 9724246.007 13322407.22 13755550.25 12684353.15 16800297.76 30040716.67 15249912.61 27092794.78 6-phosphogluconic acid 1085867.213 1640464.52 2207001.608 2615904.315 3218794.027 4075907.128 10311693.9 10296985.58 15834232.62 26361539.38 20948047.72 30124456.82 acetylcarnitine+ 48566803.84 18363645.52 212876291 19837473.54 287069457.3 179279984.4 75981677.23 68692630.65 64065401 19640895.77 124642770.2 114166825.7 acetylcholine+ 11991841.05 7988744.884 9087320.626 9100760.936 11366430.62 8688015.254 6007836.129 5327051.955 7524023.019 5459859.96 4745009.824 5434795.357 acrylic acid 19669813.64 22941468.84 23382310.17 12261121.63 14580505.65 18835140.02 22738420.88 18425950.66 17357829.27 16991668.52 22046074.99 18719627.44 adenine 4294200.079 5966277.482 3639251.029 6084538.399 8352801.529 6056739.93 5331012.811 4709060.691 5346508.183 4809310.443 3658842.261 6095600.471 adenosine 22646507.56 14804891.8 12998135.21 6966701.883 6337404.156 14903123.96 13330824.54 7855549.022 15910413.14 4486852.649 7709675.566 7213814.877 ADP / dGDP 10809195.65 10327839.03 12665415.1 16145199.77 14859222.51 11724742.59 16058599.1 15510951.42 18757346.39 10420141.4 13207866.47 11389073.07 agmatine 7756.096152 168968.0153 45797.23021 38776.88046 42634.44844 86833.01673 268215.4937 92346.38909 67971.25107 112192.6175 72859.51092 216935.3844 alanine 13038558.25 8717349.559 17612299.12 15499880.43 16227194.08 17348381.88 16380491.6 18240223.15 16725391.47 16376946.07 16374199.4 19846544.69 alanine methyl ester 1107872.437 1081828.061 1345999.373 2524099.998 2358981.543 1355998.772 1046812.215 941457.9544 1958121.06 1750549.385 1147225.537 1093026.741 allantoin 2223826.323 3898958.164 2275888.601 2690334.272 2647938.099 3551517.503 3443694.113 2395641.986 3069023.651 3770369.426 2460536.245 3263798.735 arginine 236847419.5 727363965.2 276898155.5 307427495.5 255708872.9 507052873 459553375.6 325506619.4 609678320.3 442900460.2 385066047.4 518244787.8 asparagine 607878.6217 817064.3368 881235.3532 436152.015 656017.8266 508697.1786 1092937.116 1728484.339 1805379.9 734247.4059 649942.9455 1256304.522 aspartic acid 23401845.09 20428043.29 28400361.68 21366631.15 20243288.25 16393685.39 15657711.93 15712745.15 18464015.98 12329237.11 8785872.752 13083286.27 ATP / dGTP 68675569.56 104714404.6 83494833.14 98368073.17 99633359.01 118460561.2 104107590.9 106528957.2 206599553.8 62997016.09 84428807.87 82573151.5 betaine+ 989948355.9 858237687.6 555076048.1 526582879.2 1162880138 651268870.4 1202571043 799939793 739928407.1 604232401.1 753095535 630326170.2 butyric acid / (iso) 31261956.15 43021663.83 42438793.87 45582594.75 25250365.8 54180501.19 42256172.33 37425476.86 45406739.41 44197633.8 34979862.62 39779867.25 butyrylcarnitine+ 6668934.108 6419169.435 6246153.813 7994180.074 9118122.342 6293829.134 5144144.851 4674937.029 6894857.705 3821175.055 3928478.857 3284604.442 cAMP(2',3') / cAMP(3',5') 13128.64827 0 16449.65616 24175.53045 26073.34367 0 54237.97604 33536.10286 47748.33299 27766.74766 16099.69563 0 carnitine+ 4170254.832 4031650.737 5921851.414 4395997.974 6174659.738 4257794.83 4353637.949 3085229.863 5092933.213 2145559.227 2286017.161 2441345.033 carnosine 0 184616.29 0 70655.24873 15479.45877 83372.72034 271009.9324 0 100503.943 78189.67426 0 127742.0573 CDP 1184815.715 1279402.345 1393328.62 2930594.26 2523439.243 2850362.956 3476710.92 2925374.193 4758049.909 3292266.193 2557399.836 3251806.366 choline+ 693809950.9 706972433.4 834929473 579220247.1 846676964.1 606613442.8 499625851.2 499924037.7 614198422.7 510798134.9 496292041 446145577.7 citraconic acid 6680654.748 10012832.03 18923731.98 21769005.98 8782257.877 9790185.807 5336737.785 9291846.835 10812000.26 17479751.01 5521183.805 5283080.869 citrate / citrate(iso) 7242913.676 9471959.156 9463504.841 9799903.468 13506437.48 14534664.93 15381458.83 16526251.23 28602487.83 8254592.264 11418410.84 14000090.14 citrulline 480449.2386 509251.2932 737238.3501 648392.6921 563536.8722 221316.1142 347366.3652 522234.4785 395829.6275 691217.8955 806566.7298 592935.8779 creatine 51901180.88 49248921.97 81568159.17 54584741.32 78581589.37 70346731.32 69379457.17 68845209.03 80978897.07 58898245.91 60117265.2 69784270.34 creatinine 97060933.96 122143405.7 87318875.59 100296574.3 117476689.2 105995058.5 94724660.96 81458779.16 92537682.85 99877750.2 85681216.64 94929973.12 CTP 7097934.444 11760118.19 7631521.39 13580566.73 15126137.67 22261012.96 15856069.56 14796764.27 36312835.04 16486953.95 16232040.45 18014547.87 cystathionine 2882418.502 3705033.097 3029944.019 2604192.621 2708616.346 3291391.895 2762251.195 2987430.204 5067310.732 2306002.99 2879134.504 2499018.866 cystine 63111.07451 66838.23678 65430.46536 172479.0843 108394.7057 141150.2758 116595.9593 369658.8511 301287.0202 657420.9689 456102.1212 633557.2778 cytidine 0 81865.06777 0 9967.67481 0 13906.19378 38495.70468 47717.36977 136007.3804 250294.8955 34058.75378 185097.4222 cytosine 842285.0896 808794.4281 1198941.954 1251663.783 915068.851 567143.2023 758732.8141 776307.5485 853686.1753 876523.6389 749444.4104 627305.5763 dATP 153189.4377 277063.8523 305278.9809 41085.9843 61844.70706 141590.876 8209.268334 13657.9653 56145.38729 15987.36837 0 0 dCTP 261327.4214 511130.924 395325.9616 268061.0166 262284.7141 429246.7313 244922.7304 196631.5226 442047.1751 79194.17351 55157.47773 84667.2579 deoxyadenosine 93458.15856 125614.0413 154522.2259 8790.061383 39122.66438 33594.33523 9561.669441 0 0 0 0 0 deoxycarnitine+ 1908873.33 2157817.551 2923401.792 1796555.985 2211993.199 1556919.268 2017280.1 2289874.574 2716856.57 1708193.5 1700541.7 1491120.761 deoxycytidine 81461.41689 81665.8826 92318.38657 63372.59525 86799.34381 27008.57221 25574.18086 9590.381727 0 0 0 0 deoxyguanosine 161801.3771 0 2837462.039 33198.86206 95266.97676 4876989.068 3042915.74 2216394.915 4150435.658 7404416.845 25793.6099 3151088.646 deoxyinosine 290438.9521 283668.9921 337231.6127 218503.489 205479.0423 173393.0322 241417.7762 246737.6391 353636.6025 137388.5551 267595.8402 222951.7558 deoxyuridine 26171.18488 42501.74849 0 16393.78594 26326.82919 12358.92183 0 14450.37309 0 0 0 79500.71539 D-galactonic acid / D-gluconic acid 749447.7114 1289577.353 2035805.99 1618360.647 1916233.624 1241752.235 2672226.517 3629244.83 3583101.053 10834015.09 5686383.877 9507632.827 dimethylglycine 1191186.421 456730.8276 1312415.227 764956.9677 895566.865 443381.2278 979025.0827 716828.6323 1086022.51 748352.6807 739597.9667 980055.8995 dopamine / octopamine 1861165.738 2229271.761 1149383.3 1628970.574 3929371.191 3480476.202 1547770.398 3142667.765 2772913.393 730255.0239 2378101.432 2051910.981 D-sedoheptulose 131015.0588 158842.8726 129099.164 85120.21932 565524.6671 328856.976 375733.6944 204432.5922 604812.4919 222822.6823 493777.1541 608710.026 D-sedoheptulose-7-phosphate 35707.64052 57544.22807 234779.6644 68290.14547 155350.3235 120085.8583 147866.27 165163.2263 299357.8179 55100.34526 87727.89768 287360.7546 dTTP 68278.71188 107627.4375 120722.1953 222370.8187 228972.882 336514.3859 95011.23095 170166.2549 258677.6022 79382.73841 14652.25747 50818.54187 epinephrine / normetanephrine 1594092.073 1690793.127 2441926.88 1405570.508 1988695.603 1625994.342 2185697.052 1659032.259 1718628.607 2044241.358 1799735.728 1766098.296 F-1,6/2,6-DP 4512104.989 6839291.833 5884536.972 11349124.55 11291035.29 13559321.95 10930429.63 9561531.818 17753922.08 9059915.855 7215127.794 8640882.134 F6P / G1P / G6P 627366.215 550345.0428 936113.2224 615473.9888 942734.871 874750.334 901876.5616 908962.446 1461876.672 845661.7684 797376.4447 1027360.303 fructose / mannose 45748848.9 104727476.4 36057529.97 42317011.38 37553545.92 40954029.24 82799477.34 42263003.67 53529261.1 79709584.11 49925696.94 57943419.49 fumaric acid 682635.5352 199135.7207 691009.8738 388157.3132 301099.7142 473701.9139 374428.7694 198814.6429 219371.7181 317476.3831 74174.37277 118925.8473 galactosamine / glucosamine / mannosamine 25756.517 0 403103.874 0 640.6286166 133007.4535 1248.822393 0 234.6678475 224.8025509 861.934607 87012.90828 galactose / glucose 32630569.42 100328485.2 34625449.38 27210999.01 22284098.59 32908949 86573096.7 43067495.3 49968537.06 67154818.04 55513250.72 66064824.46 GalNAC / GlcNAC 0 214879.9159 264681.9848 0 0 55963.5779 0 26688.13479 103444.8772 0 39789.06675 0 GAP / DHAP 715209.4971 0 602964.4035 629553.596 691967.8989 113339.787 235720.3034 355667.3835 998876.9543 674802.4136 111363.2433 656085.0981 GDP 2385363.651 2836390.298 1971036.604 2302043.556 2160967.93 2648110.833 2365011.206 2121447.929 4824190.319 1211102.701 1787580.389 2041175.736 glutamate 154506468.5 128007906.3 199141172.5 128439440 154545626.2 91153556.08 163931837 163746853.2 197832297.3 141262093.7 135554726.8 160353371.2 glutamine 46970150.89 54443398.3 85970442.44 49641392.52 55876429.75 31572697.59 63379357.43 53748084.73 63607158.65 73590529.26 67592843.23 79420773.71 glutathione 3475959.928 1891699.821 6736579.342 3354743.316 4318888.286 1728844.726 2214022.705 3136163.889 4554785.124 1681099.207 2082539.633 1994721.103 glutathione disulfide 35379180.23 56503151.92 42863653.92 49125609.58 45537508.53 48301796.31 46886590.38 43282725.9 68799815.73 25683284.03 31798332.88 31953570.08 glyceric acid 29419370.32 40416853.19 26607036.02 26673947.87 31419118.46 29725077.32 34665347.54 30391910.28 40005366.71 44329034.16 29446382.91 39964155.69 glycine 8810744.204 9271633.561 16038046.66 10642539.59 6749659.041 4528262.341 8379176.768 8373719.368 8831945.179 8513419.677 7944509.866 7870580.967 glycylglycine 0 0 161266.1183 242431.1115 253326.7182 0 0 337443.1949 204262.4189 0 134159.287 0 glyoxylic acid 1285325.248 4348041.114 1532572.68 1811274.501 1730268.745 1458822.188 2557359.887 2686121.108 2025266.902 4954780.547 1396239.087 2599117.965 GTP 11953706.4 19065241.56 11415502.04 17629593.47 17387756.13 24568077.85 17757161.65 13529818.89 37828711.02 11492734.73 12543943.63 13347861.33 guanidine 6423670.119 1490147.005 14191670.14 1188841.672 22194897.16 4517491.626 6152124.667 1176565.174 3224607.169 11966509.61 16667855.71 9271648.033 guanine 143327.4358 194452.9411 121919.612 2210085.226 193495.5406 110519.5323 173201.2266 296760.6106 155027.979 180038.9636 67421.50817 539466.548 guanosine 360802.959 96450.61595 384302.3668 41397.39052 153251.2372 277113.7233 265837.8779 150998.0239 267378.5219 25274.73563 161418.6774 271067.4905 hexanoylcarnitine+ 1370972.589 1075790.787 1239096.075 1140400.638 1266582.395 1225159.484 1442498.408 1096813.135 1311135.491 1221427.627 1000140.241 1039514.538 histidine 15317664.61 18163626.21 18503236.83 15301935.01 16542191.38 19296120.75 19723688.74 18973926.67 26995510.37 18591756.3 19768202.33 21440301.81 homoserine / threonine / allothreonine 44559749.24 47062940.17 65141845.93 41347835.7 47899603.71 40694225.12 64982268.97 55088195.88 66658332.65 66883739.45 57267855.07 62393829.14 hydroxyphenyllactic acid / HVA 864707.2919 997210.2465 1024200.826 1036158.137 1051501.997 963157.8543 1164798.096 1037612.835 1174995.486 1362151.159 975359.3431 1059292.929 hypoxanthine 6064156.594 14680434.79 6861606.376 14056849.72 9269543.004 10761148.9 11298488.96 6923980.516 9124195.272 14113452.26 10610591.24 21005150.03 inosine 57538.8002 45867.10298 37921.99905 39012.09718 0 53435.9968 94361.43332 52715.67931 145675.0297 48138.24817 83199.91592 366481.3795 kynurenic acid 13222.70166 79498.16562 0 1547606.509 18504.70672 21040.59431 57073.63747 15225.6866 0 27101.70017 15207.7408 56433.21504 kynurenine 286448.0519 797012.6786 451777.3656 548357.2457 394821.2339 749746.7564 587963.9507 434168.6743 695512.9343 692242.8468 455359.4031 582529.4637 lactic acid 14557876653 15071207150 5197283669 13421070324 14831683160 6322996777 17561971579 13469384097 15351524457 12597091383 15232402054 13520618044 lactose / maltose 0 0 0 0 187143.7086 82723.11826 87602.75284 0 111991.8255 0 0 0 L-arginino-succinate 103064.1935 14135.30252 200724.2191 32343.70823 81356.47311 110648.6967 88331.58001 36188.94915 94525.06903 76314.06154 46050.96344 22793.2217 leucine / leucine(iso) / norleucine 484432711.9 1030281601 783173104.9 505459866 438498099.9 591942974.6 761477758.3 426112441 880109610.8 1060636802 676377898 1173370452 levulinic acid 7323952.922 6848125.04 7286732.731 5373894.925 6749804.678 7332416.144 6113243.901 11542591.52 7507490.353 8964821.602 4919349.021 6080940.936 L-NMMA 1375767.428 4451966.465 1433442.886 1440059.898 1392089.171 3485303.816 2692454.954 1970365.479 3860173.969 2350149.4 2198156.829 4575713.018 L-tyrosine methyl ester 992155.9291 1101737.347 449722.4131 441439.3077 923186.0056 1238682.353 1102481.835 889272.9542 1110678.737 911292.4336 1135833.204 1103240.783 lysine 121668689.3 411688691.6 139648175.5 160192315.7 123218636 249429254 261315874.7 147824911.7 314072538.8 291900892.4 181834120.9 246962073.6 maleic acid 12130615.04 22700999.28 33687678.15 26434224.34 10311590.89 19932569.49 13097407.09 15723973.86 16610627.54 25830653.29 11906391.59 14048883.1 malic acid 9903986.898 7217859.618 9119869.237 8867764.639 9575671.383 4737531.24 6274828.424 7709814.175 8175942.707 7929687.48 5440564.582 9308817.584 ManNAC 305339.702 384513.5646 447580.3381 289408.4503 346040.124 223656.6869 436078.1087 460952.7708 331114.0774 274310.0509 231666.8472 283280.7904 methionine 184172568.3 290068283.5 198647984 180994714 172865409.7 175265640.5 332152079.9 290605125.2 250838103.2 262994201.1 241492542.6 221958294.4 methylglyoxal 3300718.522 3501212.713 4871484.398 3221480.547 5689461.812 3208728.771 2656268.316 3149097.28 2918268.135 2983701.143 2851025.135 2720258.795 methylmalonic acid 28929317.28 33201908.44 19699070.34 31277519.65 44619908.72 23641516.58 44850218.83 35037856.03 53441063.69 56078698.27 17437749.8 53102563 N1/N8-acetylspermidine 280863.8406 1824801.444 8350238.302 1642240.4 6095891.581 4417077.339 3718217.481 5821554.807 5426918.834 3681194.806 11639082.04 11827536.85 N1-acetylspermine 62906.80221 63055.50076 93328.49314 11894.05207 26557.58432 11841.56484 211774.9484 0 27894.40704 0 0 0 N-acetylaspartic acid 758335.2345 619871.2198 1518505.758 924185.8926 1544827.849 679859.2847 970628.195 1563368.369 1656689.722 1017728.096 974498.5646 1302028.612 N-acetyl-b-alanine / N-acetyl-alanine 5397640.311 13246601.21 8632460.81 11190123.25 7819113.757 11474323.88 13727501.86 8423535.54 15891290.25 14634785.99 8603116.921 12507616.83 N-acetyl-DL-serine / O-acetyl-L-serine 8139090.667 11115035 13264950.79 8404179.138 19350813.16 14191085 14073385.51 11460349.34 15319254.64 17311451.15 8993631.33 15615891.7 N-acetylglycine 11950766.76 26102078.76 8199090.207 18528873.72 17750889.53 18960212.27 28705648.42 23202295.84 27481754.01 36940577.99 17674733.97 31786694.1 NADP+ 646003.7993 610795.5297 526928.2333 642138.0086 664936.9782 809705.3062 877672.1009 1049577.93 1785941.466 700636.7693 916046.1451 1055172.958 N-formyl-L-methionine 2108738.529 2299392.133 2064981.434 1439633.009 2104399.517 1981842.652 2028192.644 1072645.3 2357637.692 987240.0901 1115081.19 821205.1176 nicotinamide 13092542.65 34427244.36 26019910.16 28095778.71 40500437.9 13159961.98 26059927.25 37144623.91 23978073.82 13119023.68 24405884.08 26104692.04 N-methylaspartic acid 27488947.25 23561738.8 50835227.21 21977031.32 30141917.45 15691956.35 28294881.67 29429126.57 34122573.4 24041812.53 22453404.56 22889959.17 norepinephrine / pyridoxine 240082436.5 517338816.6 335816651.7 538345035.7 198701829.1 464913308.2 467992252.8 325251593.4 429380311.9 518658642.6 423499289.5 523174189 ornithine 11160777.26 19311681.65 7077880.812 8497638.733 7534997.721 12057204.9 21399984.91 9813772.224 18411481.83 10991314.43 9580538.928 14274763.78 orotic acid 610744.0471 906496.6411 2384565.58 71133.19356 225641.7847 0 0 42979.86912 0 0 0 0 palmitic acid 6877717619 5250019908 10817754511 4910599868 7104824320 7774085240 7299135675 9399138651 8617205394 5422443160 5626128983 5361985761 pantothenic acid 43920564.32 57622650.76 148887358.1 60986892.02 86654837.88 56680192.13 70722301.79 51459365.4 57412515.29 65526965.6 43017455.34 46538260.18 phenylalanine 134903990.4 272595931.7 173765152.8 158189761.5 159846344.9 171313495.6 234708965.5 153050939.9 247824362.2 295920762.6 216614046.3 286621340.4 phosphocholine+ 1339198413 1715095735 1216396104 1438585041 1474548246 1787477462 1358863053 1336349935 2382653736 862661136.1 953351271.9 973698168.3 phosphocreatine 37702000.79 26627375.77 38356434.43 24943931.17 30306339.11 18760980.91 21715371.41 20047269.16 25304937.92 13384985.72 11341128.99 13739001.66 picolinic acid 807714.3108 3262942.921 951460.7934 1436379.201 808475.1912 1414849.475 1099819.254 1308858.419 1629377.598 1910023.559 2370084.816 1648856.868 proline 144671995.8 152254747.2 527571743.3 84119263.41 226472178.4 177303335.7 190523280.7 175692664.1 191056896.4 39415228.9 123356746.2 121600663.5 propionylcarnitine+ 13166544.29 10144617.71 102456747.4 11842947.37 44479778.61 19118637.67 13195316.88 5403414.049 8404228.563 2165837.17 4649212.994 4256673.742 pseudouridine 46485.30252 92002.09288 693917.4471 167620.9573 48164.56996 158538.8542 297246.6829 221777.2241 570701.4869 385290.3842 137636.7782 174103.583 purine 10652.81521 16671.98537 0 11527.10778 4566.762777 12195.65307 22740.89784 8852.910828 0 12506.53673 22585.48035 10365.91466 putrescine 1236618.35 20394538.32 51386491.9 10446704.8 35121129.27 29352891.8 22280851.18 14586568.64 14487595.28 10405402.98 9369465.059 11907680.65 pyridoxal 3609629.257 4482339.328 4855619.067 16577805.81 6116046.6 7157608.175 6275387.571 9742131.416 9024867.196 9738247.92 7395087.054 10249394.39 pyroglutamic acid 48286137.41 190427012.7 99734016.69 194156958.4 110009413.7 151482078.1 298265747.8 249760531.4 374415742 475641429.3 130072653.4 424151546.4 pyrroline hydroxycarboxylic acid 11273184.93 11794713.26 19549183.85 12055942.89 12867547.91 7691841.897 16072986.02 13479630.6 16498576.18 16812253.59 14215178.61 17812027.51 pyruvic acid 35177250.37 32162143.77 26770776.8 38268287.22 32457682.39 45938967.03 28784218.94 30856032.81 36525353.39 21047202.7 25219418.94 39494130.07 S-adenosyl-L-methionine+ 7635513.967 10656103.66 7358022.584 8143362.358 9727395.001 12444881.11 13709739.56 17460502.05 28006155.65 14900412.69 18618998.99 19341767.97 serine 15102140.61 17821747.23 20860196.51 14702990.01 17183915.74 14252473.94 23023084.07 21417785.89 24081246.81 23448434.76 21987624.46 25023576.96 sorbitol / galactitol / L-iditol / manitol 31342018.43 103095413.6 22786207.97 74996089.27 94439367.62 70518885.98 89628650.43 63075829.82 67695769.08 190123274.5 76382011.12 73043452.02 spermidine 188979023.1 209277502.4 711074898.3 121615615.6 440423406.5 258447095.1 171357767.5 125547003.2 102894835.4 50171354.09 94390133.37 79363516.65 spermine 14169745.92 16353077.28 29591538.31 13085751.67 25734093.33 19473620.98 22380205.82 18775720.16 18762561.68 13811288.39 21594522.33 18650945.53 succinic acid 2039841.383 6019844.889 2398443.029 3205987.856 1561607.836 5016538.221 5253928.703 3801053.175 2529408.951 5021059.947 3238293.365 3446808.986 sucrose 709696.5189 3230973.542 716193.8687 128475.3952 574711.082 1563697.226 1207327.568 3644220.457 454242.1756 0 4525519.264 1964849.566 taurine 178157191.6 190053736.3 301134792.3 104573230.1 271732604.1 190798039.6 158646535.9 160480172.2 171511069.8 131655512.8 164194025.1 158556358.7 thiamine diphosphate+ 64941.44698 73862.33884 58530.36045 65118.01169 81745.85529 174472.8541 100820.8677 84642.79971 274698.6769 53265.41426 78145.32409 108473.1148 thiamine phosphate+ 0 0 0 8261.519487 7798.899309 7073.90476 7402.905979 7264.252246 39169.39253 0 13047.3281 28325.25735 thiamine+ 14265702.26 10569045.74 313175949.1 5849169.819 170529806.1 105085823.4 22284832.34 17075320.09 23439090.3 6864605.675 32399544.38 89905633.3 thymine 3435816.119 1641425.27 2134860.39 1270793.261 6884739.018 3992345.182 6190916.814 1930199.255 1711229.838 2864473.642 6392118.587 3727024.151 tryptamine 551981.7705 432968.1821 462474.5906 530435.2285 623516.6839 474135.2581 739283.8601 732264.6461 589697.1654 451603.6138 431886.6317 582604.3578 tryptophan 25491281.84 45823603.58 34494420.79 30829052.55 28018599.7 30869744.61 39958392.24 26458916.8 42450274.87 46187700.7 31951936.38 48673081.34 tyramine 88525.36299 121794.5695 186780.4603 105850.8217 262702.6343 407074.8713 119976.1697 233884.1431 210455.9748 191342.0366 142138.186 488768.9593 tyrosine 117441520.9 165584552.9 224721112.5 77489706.24 196452724.1 194024852.7 213527590.5 151440322.7 168727241.3 149841460.5 189911567.8 224454451 UDP 2786844.148 2759338.936 3388351.25 5381673.401 5466080.838 4738237.622 6087885.401 5149051.864 7366012.243 4400650.824 4904971.093 6535983.03 UDP-D-galactose / UDP-D-glucose 135704.6377 88503.32223 265511.1118 188735.0569 390560.3503 262950.2262 613362.4832 391992.3844 1102039.219 605169.3497 193801.1565 533047.382 uracil 3943519.346 7158063.566 4656763.99 4140271.11 3015972.686 4663803.358 6652495.038 4506501.782 5746541.009 6982956.066 4871849.992 7248236.638 urea 21714443.43 26317557.56 24359582.17 27702149.31 34325316.64 33397852.77 41968842.56 30871041.57 31888604.34 33044307.31 29313644.07 34722872.38 uric acid 159641.6302 1076630.776 1057476.957 2500861.438 355347.7817 224995.0946 522143.4177 270839.8645 126031.9508 179148.7257 385229.8146 141377.0206 uridine 270252.2334 272929.8929 121922.7541 130327.8892 76359.96313 297998.1803 233773.6928 91057.02022 324386.3518 148774.1802 163137.3296 198952.2788 UTP 3616056.557 6033419.928 4622264.278 7215219.848 8059332.07 10653817.33 9487189.731 7819586.695 15948107.64 7079697.527 7032058.223 8331751.835 valine / norvaline 170848613.4 380341469.4 197154556.5 137080824.3 199041777.4 209133388.6 332539273.8 227138337.9 319425634 335521043 275977795.5 311519102.9 vitamin B2 97817.69797 153660.0361 143297.2892 140101.4179 123641.0166 137991.2395 186783.553 98200.56658 156575.6466 271021.5101 183583.9409 178454.9396 xanthine 486595.7409 1046091.779 584363.8167 745054.4301 501967.2169 666947.9207 808530.835 436839.3044 692611.5142 921947.4528 608469.2528 800731.0712 xylitol / arabitol / ribito 1853922.123 4414220.286 2724323.569 1578512.435 2366405.304 2468249.955 2455210.84 3236943.379 2747966.539 3214520.702 3250891.05 4340721.507 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name 1-methylnicotinamide+ C02918 1-phenylethylamine C02455 2- / 3-phosphoglycerate C00631 / C00197 2,3-bisphospho-D-glycerate C01159-C03339 2-/ 3- hydroxybutanoic acid C05984 / C01089 2-aminobutyric / 2-/3-aminoisobutyric acid C02261 / C03665 / C05145 2-HG C02630 2-ketovaleric acid C06255 3- / 4-hydroxyphenylacetic acid C05593 / C00642 3- / 7-methylguanine C02230 / C02242 3-phospho-serine C01005 4-hydroxybutyric acid C00989 4-hydroxyproline / 5-aminolevulinate C01157 / C00430 5-HIAA C05635 5-methylcytosine C02376 5'-methylthioadenosine C00170 6-phosphogluconic acid C00345 acetylcarnitine+ C02571 acetylcholine+ C01996 acrylic acid C00511 adenine C00147 adenosine C00212 ADP / dGDP C00008 / C00361 agmatine C00179 alanine C00041 alanine methyl ester n/a allantoin C01551 arginine C02385  asparagine C16438 aspartic acid C16433 ATP / dGTP C00002 / C00286 betaine+ C00719 butyric acid / (iso) C00246 / C02632 butyrylcarnitine+ C02862 cAMP(2',3') / cAMP(3',5') C02353 / C00575 carnitine+ C00487 carnosine C00386 CDP C00112 choline+ C00114 citraconic acid C02226 citrate / citrate(iso) C00158 / C00311 citrulline C00327 creatine C00300 creatinine C00791 CTP C00063 cystathionine C00542 cystine C01420 cytidine C00475 cytosine C00380 dATP C00131 dCTP C00458 deoxyadenosine C00559 deoxycarnitine+ C01181 deoxycytidine C00881 deoxyguanosine C00330 deoxyinosine C05512 deoxyuridine C00526 D-galactonic acid / D-gluconic acid C00880 / C00257 dimethylglycine C01026 dopamine / octopamine C03758 / C04227 D-sedoheptulose C02076 D-sedoheptulose-7-phosphate C05382 dTTP C00459 epinephrine / normetanephrine C00788 / C05589 F-1,6/2,6-DP C00354 / C00665 F6P / G1P / G6P C00085 / C00103 / C00092 fructose / mannose C00095 / C00159 fumaric acid C00122 galactosamine / glucosamine / mannosamine C02262 / C00329 / C03570 galactose / glucose C00124 / C00267 GalNAC / GlcNAC C01074 / C00140 GAP / DHAP C00118 / C00111 GDP C00035 glutamate C00302 glutamine C00303 glutathione C00051 glutathione disulfide C00127 glyceric acid C00258 glycine C00037 glycylglycine C02037 glyoxylic acid C00048 GTP C00044 guanidine C17349 guanine C00242 guanosine C00387 hexanoylcarnitine+ n/a histidine C00768 homoserine / threonine / allothreonine C00263 / C00188 / C05519 hydroxyphenyllactic acid / HVA C03672 / C05582 hypoxanthine C00262 inosine C00294 kynurenic acid C01717 kynurenine C01718 lactic acid C01432 lactose / maltose C00243 / C00208 L-arginino-succinate C03406 leucine / leucine(iso) / norleucine C16439 / C16434 / C01933 levulinic acid n/a L-NMMA C03884 L-tyrosine methyl ester C03404 lysine C00047/C00739 maleic acid C01384 malic acid C00711 ManNAC C00645 methionine C00073 methylglyoxal C00546 methylmalonic acid C02170 N1/N8-acetylspermidine C00612 / C01029 N1-acetylspermine C02567 N-acetylaspartic acid C01042 N-acetyl-b-alanine / N-acetyl-alanine C01073 / n/a N-acetyl-DL-serine / O-acetyl-L-serine n/a / C00979 N-acetylglycine n/a NADP+ C00006 N-formyl-L-methionine C03145 nicotinamide C00153 N-methylaspartic acid C12269 norepinephrine / pyridoxine C00547 / C00314 ornithine C01602 orotic acid C00295 palmitic acid C00249 pantothenic acid C00864 phenylalanine C02057 phosphocholine+ C00588 phosphocreatine C02305 picolinic acid C10164 proline C16435 propionylcarnitine+ C03017 pseudouridine C02067 purine C15587 putrescine C00134 pyridoxal C00250 pyroglutamic acid C01879-C02237 pyrroline hydroxycarboxylic acid C04281 pyruvic acid C00022 S-adenosyl-L-methionine+ C00019 serine C00716 sorbitol / galactitol / L-iditol / manitol C00794 / C01697 / C01507 / C00392 spermidine C00315 spermine C00750 succinic acid C00042 sucrose C00089 taurine C00245 thiamine diphosphate+ C00068 thiamine phosphate+ C01081 thiamine+ C00378 thymine C00178 tryptamine C00398 tryptophan C00806 tyramine C00483 tyrosine C01536 UDP C00015 UDP-D-galactose / UDP-D-glucose C00052 / C00029 uracil C00106 urea C00086 uric acid C00366 uridine C00299 UTP C00075 valine / norvaline C16436 / C01799 vitamin B2 C00255 xanthine C00385 xylitol / arabitol / ribito C00379/ C01904 / C00532 / C00474 METABOLITES_END #END