#METABOLOMICS WORKBENCH shuibingchen_20210924_093504 DATATRACK_ID:2863 STUDY_ID:ST001921 ANALYSIS_ID:AN003122 PROJECT_ID:PR001212 VERSION 1 CREATED_ON September 27, 2021, 6:30 pm #PROJECT PR:PROJECT_TITLE An Airway Organoid-Based Screen Identifies a Role for the HIF1α-Glycolysis Axis PR:PROJECT_TITLE in SARS-CoV-2 Infection PR:PROJECT_TYPE MS PR:PROJECT_SUMMARY SARS-CoV-2 causes the COVID-19 pandemic. It is urgent to develop disease models PR:PROJECT_SUMMARY to dissect mechanisms regulating SARS-CoV-2 infection. Here, we derive airway PR:PROJECT_SUMMARY organoids from human pluripotent stem cells (hPSC-AOs). The hPSC-AOs, PR:PROJECT_SUMMARY particularly ciliated-like cells, are permissive to SARS-CoV-2 infection. Using PR:PROJECT_SUMMARY this platform, we perform a high content screen and identify GW6471, which PR:PROJECT_SUMMARY blocks SARS-CoV-2 infection. GW6471 can also block infection of the B.1.351 PR:PROJECT_SUMMARY SARS-CoV-2 variant. RNA-seq analysis suggests that GW6471 blocks SARS-CoV-2 PR:PROJECT_SUMMARY infection at least in part by inhibiting HIF1α, which is further validated by PR:PROJECT_SUMMARY chemical inhibitor and genetic perturbation targeting HIF1α. Metabolic PR:PROJECT_SUMMARY profiling identifies decreased rates of glycolysis upon GW6471 treatment, PR:PROJECT_SUMMARY consistent with transcriptome profiling. Finally, xanthohumol, PR:PROJECT_SUMMARY 5-(Tetradecyloxy)-2-furoic acid, and ND-646, three compounds that suppress fatty PR:PROJECT_SUMMARY acid biosynthesis, also block SARS-CoV-2 infection. Together, a high content PR:PROJECT_SUMMARY screen coupled with transcriptome and metabolic profiling reveals a key role of PR:PROJECT_SUMMARY the HIF1α-glycolysis axis in mediating SARS-CoV-2 infection of human airway PR:PROJECT_SUMMARY epithelium. PR:INSTITUTE Weill Cornell Medical College PR:DEPARTMENT Surgery PR:LAST_NAME Chen PR:FIRST_NAME Shuibing PR:ADDRESS A 827B, 1300 York Ave, New York, NY, 10065, USA PR:EMAIL shc2034@med.cornell.edu PR:PHONE 2127465431 #STUDY ST:STUDY_TITLE An Airway Organoid-Based Screen Identifies a Role for the HIF1α-Glycolysis Axis ST:STUDY_TITLE in SARS-CoV-2 Infection ST:STUDY_SUMMARY SARS-CoV-2 causes the COVID-19 pandemic. It is urgent to develop disease models ST:STUDY_SUMMARY to dissect mechanisms regulating SARS-CoV-2 infection. Here, we derive airway ST:STUDY_SUMMARY organoids from human pluripotent stem cells (hPSC-AOs). The hPSC-AOs, ST:STUDY_SUMMARY particularly ciliated-like cells, are permissive to SARS-CoV-2 infection. Using ST:STUDY_SUMMARY this platform, we perform a high content screen and identify GW6471, which ST:STUDY_SUMMARY blocks SARS-CoV-2 infection. GW6471 can also block infection of the B.1.351 ST:STUDY_SUMMARY SARS-CoV-2 variant. RNA-seq analysis suggests that GW6471 blocks SARS-CoV-2 ST:STUDY_SUMMARY infection at least in part by inhibiting HIF1α, which is further validated by ST:STUDY_SUMMARY chemical inhibitor and genetic perturbation targeting HIF1α. Metabolic ST:STUDY_SUMMARY profiling identifies decreased rates of glycolysis upon GW6471 treatment, ST:STUDY_SUMMARY consistent with transcriptome profiling. Finally, xanthohumol, ST:STUDY_SUMMARY 5-(Tetradecyloxy)-2-furoic acid, and ND-646, three compounds that suppress fatty ST:STUDY_SUMMARY acid biosynthesis, also block SARS-CoV-2 infection. Together, a high content ST:STUDY_SUMMARY screen coupled with transcriptome and metabolic profiling reveals a key role of ST:STUDY_SUMMARY the HIF1α-glycolysis axis in mediating SARS-CoV-2 infection of human airway ST:STUDY_SUMMARY epithelium. ST:INSTITUTE Weill Cornell Medicine ST:LAST_NAME Chen ST:FIRST_NAME Shuibing ST:ADDRESS A 827B, 1300 York Ave ST:EMAIL shc2034@med.cornell.edu ST:PHONE 2127465431 #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 - Mock1 Treatment:mock RAW_FILE_NAME=Mock1.raw SUBJECT_SAMPLE_FACTORS - Mock2 Treatment:mock RAW_FILE_NAME=Mock2.raw SUBJECT_SAMPLE_FACTORS - Mock3 Treatment:mock RAW_FILE_NAME=Mock3.raw SUBJECT_SAMPLE_FACTORS - Mock4 Treatment:mock RAW_FILE_NAME=Mock4.raw SUBJECT_SAMPLE_FACTORS - SARS_CoV_2_1 Treatment:SARS_CoV_2 RAW_FILE_NAME=SARS_CoV_1_13_23.raw SUBJECT_SAMPLE_FACTORS - SARS_CoV_2_2 Treatment:SARS_CoV_2 RAW_FILE_NAME=SARS_CoV_2_11_22.raw SUBJECT_SAMPLE_FACTORS - SARS_CoV_2_3 Treatment:SARS_CoV_2 RAW_FILE_NAME=SARS_CoV_1_12.raw SUBJECT_SAMPLE_FACTORS - SARS_CoV_2_4 Treatment:SARS_CoV_2 RAW_FILE_NAME=SARS_CoV_2_21.raw SUBJECT_SAMPLE_FACTORS - GW+SARS_CoV_2_1 Treatment:GW+SARS_CoV_2 RAW_FILE_NAME=GW1.raw SUBJECT_SAMPLE_FACTORS - GW+SARS_CoV_2_2 Treatment:GW+SARS_CoV_2 RAW_FILE_NAME=GW2.raw SUBJECT_SAMPLE_FACTORS - GW+SARS_CoV_2_3 Treatment:GW+SARS_CoV_2 RAW_FILE_NAME=GW3.raw SUBJECT_SAMPLE_FACTORS - scramble shRNA+SARS+CoV-2_1 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=scramble shRNA_1.raw SUBJECT_SAMPLE_FACTORS - scramble shRNA+SARS+CoV-2_2 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=scramble shRNA_2.raw SUBJECT_SAMPLE_FACTORS - scramble shRNA+SARS+CoV-2_3 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=scramble shRNA_3.raw SUBJECT_SAMPLE_FACTORS - shHIF1+SARS+CoV-2_1 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=shHIF1_1.raw SUBJECT_SAMPLE_FACTORS - shHIF1+SARS+CoV-2_2 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=shHIF1_2.raw SUBJECT_SAMPLE_FACTORS - shHIF1+SARS+CoV-2_3 Treatment:scramble shRNA+SARS+CoV-2 RAW_FILE_NAME=shHIF1_3.raw #COLLECTION CO:COLLECTION_SUMMARY hPSC-AOs were collected into 150 mM ammonium acetate. After centrifugation, cell CO:COLLECTION_SUMMARY pellets were collected for metabolite profiling. Metabolite extraction was CO:COLLECTION_SUMMARY performed with 200 μl of 80% methanol (20% water) with internal standards (U13C CO:COLLECTION_SUMMARY succinate, and U13C citrate, and heptadecanoate). CO:SAMPLE_TYPE Lung organoids #TREATMENT TR:TREATMENT_SUMMARY SARS-CoV-2 variants, including USA-WA1/2020 and B.1.351, were obtained from the TR:TREATMENT_SUMMARY World Reference Center for Emerging Viruses and Arboviruses located at the TR:TREATMENT_SUMMARY University of Texas, Medical Branch via the CDC. SARS-CoV-2 was propagated in TR:TREATMENT_SUMMARY Vero E6 cells (ATCC) in EMEM supplemented with 10% FCS, 1 mM Sodium Pyruvate and TR:TREATMENT_SUMMARY 10 mM HEPES. hPSC-AOs were fragmented into small cell clusters and plated on 10% TR:TREATMENT_SUMMARY matrigel-coated plates. The infection was performed in the culture media at the TR:TREATMENT_SUMMARY indicated MOIs at 37°C. For pre-infection treatment experiments, hPSC-AOs were TR:TREATMENT_SUMMARY pretreated with DMSO (control), 10 µM GW6471 for 4 hours prior to infection. TR:TREATMENT_SUMMARY HIF1a knockdown in hPSC-AOs Two shRNAs against HIF1α and one scrambled negative TR:TREATMENT_SUMMARY control non-effective shRNA in lentiviral GFP vectors were purchased from TR:TREATMENT_SUMMARY OriGene company. The shRNA sequences are shown in Table S3. To generate TR:TREATMENT_SUMMARY lentivirus particles, shRNA vectors and lentivirus packaging vectors were TR:TREATMENT_SUMMARY co-transfected into 293T cells. The day 15 lung progenitor cells were infected TR:TREATMENT_SUMMARY with the collected lentivirus (MOI=0.5) with 8 μg/ml polybrene. To increase the TR:TREATMENT_SUMMARY infection efficiency, the cells were centrifuged at 2300 rpm for 1 hour at TR:TREATMENT_SUMMARY 30℃. At 24 hpi, the cells were selected with 1 μg/ml puromycin for an TR:TREATMENT_SUMMARY additional 72 hours. After selection, the cells were dissociated into single TR:TREATMENT_SUMMARY cells and seeded into 24 well plates at 300-400 cells/μl for 3D organoid TR:TREATMENT_SUMMARY differentiation. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY In brief, the samples were methoximized with 50 µl of methoxyamine SP:SAMPLEPREP_SUMMARY hydrochloride (MOA, 15 mg/mL in pyridine) at 30°C for 90 minutes. The SP:SAMPLEPREP_SUMMARY silylation step was done with 50 µl of SP:SAMPLEPREP_SUMMARY N,O-Bis(trimethylsilyl)trifluoroacetamide (BSTFA, containing 1% TMCS, Supelco, SP:SAMPLEPREP_SUMMARY PA, USA) at 70 °C for 60 minutes. After the samples cooled to room temperature, SP:SAMPLEPREP_SUMMARY they were analyzed by GC-TOF/MS. Separation was performed with a 30-meter DB-5MS SP:SAMPLEPREP_SUMMARY column coupled with 10-meter guard column. Helium was used as carrier gas at a SP:SAMPLEPREP_SUMMARY flow of 1 ml/min. The oven program is as the following: started at 60 °C for 1 SP:SAMPLEPREP_SUMMARY minute, 10°C /min to 320 minutes and kept for 3 minutes. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Separation was performed with a 30-meter DB-5MS column coupled with 10-meter CH:CHROMATOGRAPHY_SUMMARY guard column. Helium was used as carrier gas at a flow of 1 ml/min. The oven CH:CHROMATOGRAPHY_SUMMARY program is as the following: started at 60 °C for 1 minute, 10°C /min to 320 CH:CHROMATOGRAPHY_SUMMARY minutes and kept for 3 minutes. CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890A CH:COLUMN_NAME 30-meter DB-5MS #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME new MS:INSTRUMENT_TYPE GC-TOF MS:MS_TYPE EI MS:ION_MODE POSITIVE MS:MS_COMMENTS The transfer line temperature was set to 250 °C. The data was acquired with MS:MS_COMMENTS MassLynx software (Waters). The data was collected with the scan time of 0.3 s MS:MS_COMMENTS and interscan delay of 0.1 s. The mass range was set to 35 to 650 for EI. Raw MS:MS_COMMENTS data files (.raw) generated from GC-TOF/MS were analyzed in Refiner MS (Genedata MS:MS_COMMENTS Expressionist, Basel, Switzerland) software. The output from Refiner MS contains MS:MS_COMMENTS retention time, quantification mass, and peak volume for each metabolite. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Area MS_METABOLITE_DATA_START Samples Mock1 Mock2 Mock3 Mock4 SARS_CoV_2_1 SARS_CoV_2_2 SARS_CoV_2_3 SARS_CoV_2_4 GW+SARS_CoV_2_1 GW+SARS_CoV_2_2 GW+SARS_CoV_2_3 scramble shRNA+SARS+CoV-2_1 scramble shRNA+SARS+CoV-2_2 scramble shRNA+SARS+CoV-2_3 shHIF1+SARS+CoV-2_1 shHIF1+SARS+CoV-2_2 shHIF1+SARS+CoV-2_3 Factors Treatment:mock Treatment:mock Treatment:mock Treatment:mock Treatment:SARS_CoV_2 Treatment:SARS_CoV_2 Treatment:SARS_CoV_2 Treatment:SARS_CoV_2 Treatment:GW+SARS_CoV_2 Treatment:GW+SARS_CoV_2 Treatment:GW+SARS_CoV_2 Treatment:scramble shRNA+SARS+CoV-2 Treatment:scramble shRNA+SARS+CoV-2 Treatment:scramble shRNA+SARS+CoV-2 Treatment:scramble shRNA+SARS+CoV-2 Treatment:scramble shRNA+SARS+CoV-2 Treatment:scramble shRNA+SARS+CoV-2 1-hexadecanol 21.94518089 23.47255325 23.03432083 22.75930023 28.98411369 34.57345963 32.84506989 32.2076416 35.71912384 31.26770782 33.6843338 16.50672913 19.69475555 21.68865585 30.13063812 28.15970802 31.25455284 1-Methylhistamine 142.7700348 139.3828735 147.1497345 162.0018311 169.3263855 174.8140564 153.8820953 174.8413696 155.6588287 150.3664551 158.2719116 9.655492783 106.8883743 114.2323303 127.6736908 128.4468689 134.4712372 1-Monopalmitin 10144.91016 10874.01465 10257.83203 10707.29395 10949.33203 12112.94141 10745.29199 11422.03125 11286.61035 9540.03125 11078.99219 67.02484131 75.67572784 67.09775543 77.98243713 70.74655914 86.3584137 2-Amino-4,6-dihydroxypyrimidine 38.28881454 37.20232391 34.43139267 35.5634346 47.00823975 55.27744675 50.73803329 57.59013367 42.53926849 48.1342659 47.21468735 986.4856567 999.4379272 985.178772 221.1237183 210.2097473 184.716095 3-Aminobutyric acid 105.2225723 89.42301941 104.6501083 86.10891724 111.76371 103.1834412 122.9446411 204.84758 101.6774521 96.34947968 92.22681427 3-hydroxyanthranilic acid 59.46465683 57.88552475 64.65167999 81.3657608 81.90068054 75.29141235 64.09204865 85.71165466 67.99118805 61.0266037 69.36223602 52.28217697 69.59975433 62.54567719 72.88058472 71.67568207 75.34780121 3-phosphoglyceric acid 3.038758278 4.050691128 4.138314724 8.920823097 91.46652222 96.2165451 68.32946014 93.52910614 13.02035332 11.51063633 16.14759827 23.63257217 119.8236542 122.2149506 139.6979065 134.7957001 147.8089447 5-aminovaleric acid 148.0089111 142.9014587 155.6152344 163.597641 183.8292694 198.1932831 182.8813171 202.3405304 180.2421875 165.8295593 176.2210083 3414.100098 4635.527832 4546.626953 4634.975098 4917.73877 5020.228027 6-phosphogluconic acid 2673.721436 2908.085938 2816.971436 2927.283936 2999.950928 3589.583496 3445.868896 3558.073975 3216.922363 2835.207764 3049.991455 1968.891479 2695.935303 2413.4646 3271.691162 3154.332275 3355.301758 6-phosphogluconic acid 2 4.531413078 2.62447691 2.852327347 6.928601265 24.20781517 30.17895889 23.09877777 25.57610893 11.17096424 9.839722633 10.9222641 5.531209946 14.25377941 13.90299606 18.12513733 19.10501099 17.66911316 Adenosine 33.76351547 38.52568817 53.71039963 51.15139389 382.0860901 437.7276611 312.5435791 438.2107849 106.4825516 81.23390961 102.2505112 146.0162811 212.6780853 152.4573975 602.7130737 249.499588 347.2800598 adipic acid 153.2018433 133.5693054 154.9349365 105.1253204 136.3769989 167.4561615 172.7825012 144.6621857 154.9499512 142.1680603 120.5895004 1859.253174 1029.436646 846.8477783 327.4103088 250.539444 234.0309753 Alanine 467.2680359 456.0508728 520.8043213 533.5380249 1502.099243 1747.89502 1478.602783 1514.101196 1292.959717 1293.986816 1306.657593 581.0043335 621.5631714 636.0273438 574.7184448 589.8510132 564.6936646 alpha tocophereol 2081.028809 2422.64502 2330.839355 2659.266846 2882.898193 3114.192871 2787.969727 3188.81543 2741.12793 2507.794189 2814.322021 1091.173706 1636.944458 1388.818359 1613.76062 1560.605591 1717.067505 alpha-glucosamine 1-phosphate 354.5110779 385.6842346 399.7531738 443.6660767 1096.398926 1351.987549 1166.461182 1206.678101 910.9493408 852.3181152 994.0174561 243.2562408 459.2666626 435.3475342 412.7002869 421.2250977 439.5604858 alpha-Hydroxyglutaric acid 7.214782715 7.387612343 7.547369957 9.094047546 26.14940834 27.56481361 24.60804367 36.51282501 23.79078674 21.21867561 24.76732635 1.384803414 6.515184402 5.203568459 12.87063313 11.64364147 11.98215389 Aspartic acid 6.682632923 22.71385384 24.91184998 32.00011826 29.6676693 30.41390228 AMP 3.17691493 6.712324619 9.024892807 35.40498352 82.0774765 39.72351837 8.613718987 57.09986115 23.4043026 18.20339584 30.29899597 21.1881218 53.73921967 48.0294342 77.83096313 80.41868591 87.91504669 benzoic acid 134.1067352 137.2972107 131.1615601 125.6429672 163.7472382 158.2618256 196.991806 252.0641785 150.7697601 155.5919037 152.9223328 144.5860748 115.1924896 121.8967514 116.8999786 112.9366226 108.6368637 Beta- alanine 19.89637566 18.89089394 21.64056396 21.5837822 47.32997894 56.40027618 50.94057465 50.7761879 42.34262848 37.8719101 44.15885925 45.94033813 79.32475281 87.11515045 32.18991852 41.44342804 42.55596542 beta-glycerolphosphate 6.230247974 20.83065033 22.92136192 44.1598587 47.97638702 53.47249222 cellobiose 6.723642826 6.289966583 5.93464613 6.359274864 92.75237274 83.02861786 46.80947495 120.408493 3.802708149 2.614938021 4.285416126 6.230247974 20.83065033 22.92136192 44.1598587 47.97638702 53.47249222 cholesterol 425.6632385 613.8762817 603.0803833 1357.686279 2136.826416 861.6233521 1022.881714 3101.896484 1302.759521 1265.846313 1373.165527 25.8045311 39.53659058 36.90834045 932.5370483 59.01074982 65.69909668 citric acid 58.65986252 71.91702271 71.105896 72.780159 131.8148804 164.4739838 141.7266693 151.6131897 57.75969315 50.85571289 57.99455643 704.2396851 694.7952881 709.4529419 675.9567261 676.9213867 681.2084351 cytindine-5'-monophosphate 29.73347092 35.07413864 33.72724152 41.9319725 70.9417572 94.12103271 74.7931366 68.72969818 49.17475891 42.01292038 48.13315582 405.0322266 411.9925537 437.1722717 365.1072998 351.7658997 357.7157288 dehydroascorbic acid 64.81704712 71.60881042 82.89652252 94.14556885 1084.808105 1221.679077 1158.239624 896.7888794 954.3151245 1017.931091 1064.597168 21.96362114 35.36870956 35.45662308 64.75095367 66.55970764 72.27174377 D-glucose-6-phosphate 2.166045666 3.217763901 3.518380642 4.264277935 110.8035507 130.184021 95.68034363 122.6445465 38.51681519 38.60667801 39.64842606 498.1213074 1113.544312 805.8623657 1095.774536 1259.35437 1415.931519 D-lyxose 203.5783234 249.1169739 255.4903717 246.7227478 460.6687927 598.1693115 510.2426453 535.0092773 307.4519653 276.3594971 315.6880493 54.99593067 58.77394485 51.67552376 43.40759277 44.33268738 46.98197556 D-malic acid 150.4456482 181.497757 159.5050659 177.1096039 794.038269 886.6552124 809.9432983 934.7451782 953.7597656 847.9904785 923.428772 D-Ribose 8.889533043 7.912572384 9.483938217 10.53133488 32.50519943 35.09908295 35.71858215 32.14836502 51.51924133 20.01665688 23.22650719 9.846031189 18.58539009 19.77205849 26.70471001 28.47557831 30.2232132 fumaric acid 38.82698822 39.16892624 42.57244492 44.59555817 74.97301483 71.13579559 69.5888443 148.5098419 42.4344902 38.59648895 37.75363541 37.76753235 39.318573 35.43731308 40.21277618 33.56946182 35.0721817 gluconic acid 2 2.288188457 3.50980854 9.656915665 5.906890869 39.90209198 53.23514938 40.28302002 45.18892288 23.09275055 20.43235016 18.16757965 gluconic acid lactone 10.84955692 14.63560486 15.24881744 16.37981796 146.4735718 193.1514893 124.2010193 170.2247467 95.79308319 74.81587982 116.9096146 100.7878571 201.704422 238.1390076 338.0795288 359.7640991 380.9099121 gluconic acid lactone 2 2.454916 2.873922825 3.737664461 4.779974937 80.80343628 84.48417664 40.63828659 80.92318726 37.58109283 31.59000587 49.29950333 29.0164299 151.9794464 152.7043915 198.9210052 246.9427948 271.8854065 Glucose 2 20.37495041 27.72774696 26.33833504 31.83724403 497.3691711 379.3775024 389.2548218 725.774231 299.5360718 300.555603 351.4122009 11107.10059 11822.44531 11892.01855 11690.55469 11799.38574 11759.57031 Gluose 157.2160492 200.1181335 188.2483978 243.5348053 2903.88916 2318.835693 2343.344971 3591.338135 1944.956299 1934.633667 2185.956055 6680.598145 7558.943359 7495.30127 7579.103516 7624.749023 7813.300293 Glutaric acid 237.8654633 225.8131409 239.8476868 254.2090149 292.7919312 281.0269775 268.6540833 480.9683838 245.5140228 241.7780457 239.4767609 93.42247009 126.6927948 193.0367737 209.0170898 212.2706757 223.5978241 glutamic acid 0.936794996 20.42605782 90.29183197 85.56424713 150.6125336 160.5918274 Glyceraldehyde 2201.215576 2202.380371 2412.178955 2904.613281 3068.775146 1987.46106 2132.849854 3314.991211 2652.26001 3045.07959 2677.994385 137.616745 111.9124146 122.2123108 144.7111969 138.1731567 149.8427277 Glyceraldehyde-3-phosphate 196.1207428 246.3088074 244.8813171 424.1786804 945.8357544 902.3764648 656.6998901 963.5095215 1051.205811 855.25 1212.718262 676.7947998 1254.768311 1141.628296 1369.756958 1353.897827 1477.511353 glyceric acid 18.19511223 11.69910431 15.6327343 14.43218136 21.36520386 16.47495079 17.71426392 24.13873482 20.42059708 24.63678741 19.37221527 283.5123901 482.1477051 529.9259033 884.303833 1002.36377 1099.114136 glycerol 33.26797485 33.57778931 34.88726044 45.29211044 72.77835846 46.12384415 48.55038834 91.33911133 34.88669968 35.7531662 34.99113083 158.6392975 173.2763824 169.5510712 164.0822754 162.7142181 168.0099792 glycerol 1-phosphate 365.6241455 393.5166321 394.0593872 495.149231 1200.47876 1581.733887 1276.482178 1312.019043 564.6174927 529.1844482 607.4455566 132.6734619 228.016861 243.9586639 451.3509827 462.5853271 507.4628906 glycolic acid 314.1390991 549.2811279 700.2444458 714.8496704 646.7446899 617.6744385 Glycine 62.454422 65.9810791 71.52875519 66.89598083 81.03355408 96.0957489 80.23474121 93.35216522 48.88479233 45.35997391 45.24030685 glycine 835.5134888 929.9360352 918.8262329 935.0794678 2423.250977 2616.771729 2467.797119 2467.92334 1872.059692 1750.24707 1934.877808 403.8006592 1269.41687 1303.225586 1476.384644 1575.44397 1621.740479 Guanosine 21.03077698 27.50006485 33.46470261 42.81447983 81.32175446 77.78028107 43.54703522 92.77043152 41.47488785 27.65584373 36.49820328 11.79279613 24.41174698 17.1397953 31.86614799 32.94754791 42.9786377 heptadecanoic acid 1085.456787 1285.813721 1358.057129 1380.663452 1553.005127 1950.237671 1718.38208 2086.813477 1558.995117 1321.261963 1491.504761 hydroxyurea 52.13277817 47.51222229 52.93312073 47.30487061 53.28103256 49.29967499 47.73967361 56.10400009 46.48031235 48.45583725 43.26368332 3.720897198 12.11937523 12.37963009 20.6643486 21.60333824 21.74361038 inosine 338.1580505 316.1722412 394.4023132 468.6082764 753.6448364 742.0764771 411.2615051 796.4849243 409.3537598 342.748291 373.1748657 67.72344971 200.7455139 118.0139542 228.0943146 184.2990112 335.2306519 Isoleucine 215.3491516 233.1560211 253.3812408 215.3908081 1111.024902 1192.277588 1045.463135 1093.65625 885.0753784 936.0494995 894.8222046 532.3053894 512.7966919 571.8601379 394.5796509 495.5240479 536.4534912 Lactic acid 7.530279636 8.743924141 8.777509689 10.59751892 99.03173065 69.30117798 77.35015869 119.4670563 114.0148163 113.270546 121.2459564 9.447122574 15.47061443 13.81829453 17.46689224 14.27453136 15.72592068 lanosterol 71.91654205 87.18109131 86.73046112 119.4606476 131.6256104 137.6665802 98.77549744 140.6365967 251.9473572 218.0847931 258.0637207 77.10494995 142.146347 130.0318451 202.2430573 188.7947235 214.1040497 L-Aspartic acid 115.0352097 157.0345917 159.7690125 165.5934143 938.7491455 1057.174194 902.2597656 986.5059204 1073.956177 1029.361694 1077.975098 Lauric acid 17.84924889 19.74795914 23.2632885 24.10630226 34.41016388 39.29203796 30.6970253 37.99897766 33.01646042 25.07933426 26.31118202 204.7841339 224.7464142 218.7132111 223.3403931 235.6594391 233.2402039 L-cysteine 96.00756836 138.2958527 145.651474 119.5698929 941.2041626 1202.926392 949.9780884 970.9841919 498.8809204 486.0718384 530.8221436 107.9175491 184.3312378 226.237915 246.8942719 390.7704163 431.6244507 L-glutamic acid 112.0800781 146.4976959 165.6877899 165.7073669 1084.132446 1178.718384 1032.828125 1042.274902 563.5427246 509.1809998 581.3735962 122.9881973 171.3919983 166.6358337 209.2996826 220.4563141 219.7463989 L-glutamine 7.630421638 8.175795555 11.55764294 15.03099442 291.5219116 241.2632904 83.31999207 189.8477173 310.1470947 368.2185974 400.8055725 L-homoserine 48.82622147 50.75403976 30.57390594 51.69694519 74.19094849 66.20028687 53.04023361 168.4405823 64.86360931 55.63106155 47.61746597 L-lysine 132.257019 210.965332 221.7001343 242.3180389 1839.840332 2111.266113 1651.434692 1851.364258 1674.387695 1627.884888 1793.401733 45.80281448 326.6209412 457.7084656 319.528595 558.9889526 569.5811768 L-mimosine 176.1672974 193.3680878 188.6003876 182.1697998 230.6609497 257.8266907 226.146286 236.4976196 169.2095032 184.3547821 169.9519501 108.0601273 187.3403931 203.9208832 265.4992371 274.4161377 290.1481018 L-pyroglutamic acid 1374.24646 1502.546997 1666.134888 1593.520996 5243.029785 5641.325195 5342.839844 5400.768555 6007.856934 5546.490234 6022.859863 L-serine 735.7936401 765.2390137 770.0935669 730.4326782 2272.387695 2414.448486 2042.704224 2206.455811 1632.831909 1594.935791 1685.762085 L-Threonic acid 44.0069809 51.30657196 55.66981888 60.22514725 350.5166016 452.0992432 386.638031 401.8298645 307.2036743 266.2245483 307.5409241 L-threonine 837.2766724 956.7313232 966.2557373 863.0325317 2973.296387 3356.188232 2971.168945 3166.858398 2298.971191 2226.241211 2304.605957 L-Tyrosine 53.72291565 74.83341217 180.9309998 108.5386887 774.6578979 858.8304443 680.6278687 779.873291 675.6971436 621.4310913 700.0716553 8.755491257 153.0442352 189.3145599 146.2169952 227.5552979 248.6697083 Methionine 14.8531971 18.68581581 20.019804 20.76814461 126.1641541 155.4857635 123.0868149 140.6088867 85.0993042 75.85366058 84.82635498 2.554129601 27.68759727 91.57981873 87.33789063 163.4717255 179.6182251 methyl-beta-D-galactopyranoside 55.0603447 67.81150055 62.69121933 69.20870209 226.4103546 318.0996094 245.5705872 272.9422302 164.0242767 153.7592468 208.425354 34.27859497 71.15474701 70.1283493 64.99810028 69.80541992 69.95629883 myo-inositol 7.015070438 8.318703651 8.866225243 10.22179604 90.5528717 67.11233521 70.6633606 132.982132 59.10105133 66.94386292 65.58027649 1127.636108 1576.043091 1448.894043 1271.3573 1255.69043 1350.375244 myristic acid 421.8345337 442.5279541 457.5090027 455.2781982 345.6332397 462.2166443 385.7415466 339.2159729 296.6725464 235.0286865 236.8045502 53.05067444 80.95058441 71.74372864 157.3011169 134.0138397 140.6854248 N-acetyl-D-glucosamine 704.4411011 660.9736328 738.0062866 1113.768921 3425.712158 3375.854736 2944.729492 3132.104492 3945.384033 3454.393066 3228.602051 1290.145142 2653.510254 1195.804321 2965.955078 2873.925049 2617.600586 Nonanoic acid 2.233038664 0.831949413 1.176159263 1.265167475 1.506983995 1.423352122 1.170056581 2.603433609 1.175764441 1.125659585 1.370519996 50.20557785 42.7473793 46.66208649 73.66540527 57.22300339 127.4375153 oleic acid 142.3464203 138.0272064 132.0935669 184.952179 326.9248047 173.1782227 131.0427246 1886.610962 129.572464 124.775177 120.2122345 1750.640991 2542.958008 2391.719238 3189.153809 3131.5896 3277.253662 oleic acid 2 128.5167847 112.95755 113.1225357 156.4131775 441.6911316 156.730835 144.434906 4766.985352 103.6194534 108.6393738 102.7723618 1109.878479 1205.517273 1445.770386 998.7685547 990.6790161 1025.825684 palmitic acid 480.7460938 420.0495605 433.4427185 430.9395142 672.3728027 516.8865356 376.7648621 1996.341553 476.8641052 576.6358032 548.8685913 6715.939941 7358.92041 7557.868652 7715.223633 7785.213379 7902.970215 palmitoleic acid 323.5186768 344.0673523 352.909729 352.334137 458.2457886 635.2327271 495.1110229 457.1446228 369.61026 256.578064 276.8829346 3124.879639 3290.719482 3364.422363 2782.950928 2759.17627 2855.984619 pantothenic acid 3.020300388 3.751045942 4.321748734 6.113316059 46.09992981 60.22179794 47.28253174 92.86702728 34.60900879 30.52184105 34.98126221 130.5159454 178.3635483 180.5209045 134.3282013 133.686203 139.7186127 Phenylalanine 65.46257019 98.18367004 172.8774872 113.9910736 897.784668 1016.420105 881.0526733 919.645752 669.2979736 611.8845825 680.5706787 5.976295948 9.608165741 9.867559433 14.61908722 16.16749382 18.23192215 phosphoric acid 6632.672363 6598.418457 6718.818848 6643.52832 7025.541992 7109.558105 6857.983887 7070.364746 6885.815918 6792.208008 6887.777832 828.5079956 886.56604 933.1015625 913.0893555 1000.936096 1002.146362 Putrescine 882.5811157 985.4796143 998.6167603 996.593811 1654.442749 2038.894897 1841.694824 1853.718262 1087.962646 995.0137329 1122.911011 15.1124115 1142.313721 1038.105347 1217.995483 1278.44165 1340.17627 pyruvic acid 23.39700317 23.21367455 27.80548668 34.28987885 39.19232941 20.41667747 23.18473053 46.3624115 31.16641998 38.99526978 32.36810303 65.01144409 99.73230743 66.92456055 188.961319 182.7146301 176.5050201 Ribo-5-Hexulosonic acid 0.451511919 0.709551156 2.695970058 1.84074676 1.84210515 3.564558744 1.180987835 1.611353636 1.555094838 110.2001724 282.3839722 284.9271545 207.8572998 224.3402863 230.3575745 Ribofuranose 4.771035671 5.360201359 4.21310091 3.100814342 42.00563812 52.78129959 59.97859192 66.55548859 33.46139145 32.53316116 35.96741486 5.131417274 4.675940037 4.512505054 1.11474061 1.358250022 1.035814762 ribose 123.3799362 204.7467041 191.5134125 271.4176331 269.6949158 288.0575256 Serine 3.645362377 4.157502651 3.961832047 5.620214939 8.725662231 5.464169025 5.622208595 11.20071602 4.014017582 3.968397379 3.945365429 12.1374979 11.45302677 11.35368919 8.829297066 9.054145813 9.047938347 stearic acid 539.1099854 475.5979919 477.2695618 570.1657104 1156.228149 602.7518311 488.7183533 3520.276123 557.4026489 788.3743896 665.8452759 1718.941528 1465.438232 1485.836182 1702.435425 1790.586426 1868.109619 stigmasterol 19.29071999 25.64156723 30.12759209 46.25136566 36.99012756 39.50116348 22.99226952 45.77936172 231.2672424 205.6602631 248.3946075 25.67180061 72.11120605 58.0079422 151.4949799 143.8631744 167.2962494 succinic acid 16.95736694 15.62477112 14.49589252 18.08038139 42.12024307 34.70586395 38.8433609 128.2269287 26.37644577 24.52201653 24.6065712 26.70262146 13.71294117 11.6633606 15.92105198 13.37545395 13.87174225 threonine 4.437290668 4.153233528 13.45661545 5.096388817 20.30759621 4.117233276 21.32172394 18.38970757 12.99266815 14.45968437 1.314976573 156.8511505 327.0551147 746.1016846 979.4060059 1483.387573 1561.281128 trehalose 35.68642426 25.68936348 30.44341469 37.19742966 769.3898315 815.3508301 510.4144287 757.7978516 121.4059448 102.8107147 114.7809982 3.082870007 26.00331116 44.9272995 58.8903656 56.05340195 53.94037628 uracil 117.2485428 105.2158051 105.3003235 103.1146469 244.5387268 271.9102478 222.5113068 266.9125366 96.53269196 105.052597 111.3014908 14.43033314 20.54046631 49.84364319 61.40045166 78.09648132 83.54459381 Valine 3.719516277 4.288794041 5.266262531 6.874155998 35.65359879 20.76352882 22.50108147 38.9956131 22.51669121 24.67373848 23.40529251 46.51584625 64.8434906 56.66371536 51.26169586 51.73283005 48.95409012 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG PubChem RT m/z 1-hexadecanol C00823 2682 17.78145308 299 1-Methylhistamine C05127 3614 14.78639706 174 1-Monopalmitin 14900 23.23207417 371 2-Amino-4,6-dihydroxypyrimidine 10932 9.319149872 73 3-Aminobutyric acid C00632 86 14.94531462 282 3-hydroxyanthranilic acid C00431 138 14.26098345 174 3-phosphoglyceric acid C01879 7405 12.9160126 156 5-aminovaleric acid 91493 21.82953393 318 6-phosphogluconic acid 91493 22.90322588 387 6-phosphogluconic acid 2 91493 22.90322588 387 Adenosine C00002 60961 23.58879797 230 adipic acid C00322 196 12.72530052 73 Alanine C00041 602 7.629962708 116 alpha tocophereol 27.19487735 502 alpha-glucosamine 1-phosphate 740 16.39279228 73 alpha-Hydroxyglutaric acid 43 13.5430315 247 Aspartic acid C00049 5960 12.26806575 232 AMP C00020 6083 26.63012482 169 benzoic acid C00108 243 9.442807938 179 Beta- alanine C00099 239 11.74637914 248 beta-glycerolphosphate 92043261 15.25138625 243 cellobiose C00185 10712 23.96276937 204 cholesterol C00187 5997 27.32664953 368 citric acid C00158 311 16.26780196 278 cytindine-5'-monophosphate 18.33473308 243 dehydroascorbic acid C01041 440667 17.15805599 205 D-glucose-6-phosphate 439958 21.01317512 387 D-lyxose C00476 439240 14.70436531 73 D-malic acid C00149 525 12.47389513 147 D-Ribose C00117 439167 19.128338 315 fumaric acid C00122 444972 10.73366338 245 gluconic acid 2 C00257 752 18.11809459 73 gluconic acid lactone C00198 7027 17.68416798 217 gluconic acid lactone 2 C00198 7027 17.87140507 217 Glucose 2 C00029 5793 17.27253585 73 Gluose C00029 5793 17.07505166 73 Glutaric acid C00025 33032 14.07254938 246 glutamic acid C00026 743 11.45678127 174 Glyceraldehyde C00118 751 7.835620891 147 Glyceraldehyde-3-phosphate 729 15.90694866 299 glyceric acid C00258 752 10.85681511 147 glycerol C00093 753 9.689062546 205 glycerol 1-phosphate C00623 754 15.66084858 299 glycolic acid C00160 757 6.47451529 147 Glycine C00037 750 10.16217112 174 glycine C00037 750 10.16217112 174 Guanosine C00035 135398635 24.58572195 324 heptadecanoic acid C16536 10465 19.54372685 327 hydroxyurea C07044 3657 9.215417289 277 inosine C00081 135398641 23.10229866 217 Isoleucine C00407 6306 9.972473417 158 Lactic acid C00186 612 6.950454599 191 lanosterol C01724 246983 28.7784017 393 L-Aspartic acid C00049 5960 12.87962551 232 Lauric acid C02679 3893 14.494583 257 L-cysteine C00097 5862 13.33620048 220 L-glutamic acid C00025 33032 14.08145031 246 L-glutamine C00064 5961 15.81486931 156 L-homoserine C00263 12647 14.64640212 290 L-lysine C00047 5962 17.36371605 174 L-mimosine C04771 440473 8.689388182 147 L-pyroglutamic acid C01879 7405 12.91894696 156 L-serine C00065 5951 10.83781842 204 L-Threonic acid C03064 5460407 13.34037217 292 L-threonine C00188 6288 11.17098981 240 L-Tyrosine C00082 6057 17.53957161 218 Methionine C00073 6137 12.87870047 176 methyl-beta-D-galactopyranoside 94214 16.50149483 217 myo-inositol C00137 892 18.99063163 305 myristic acid C06424 11005 16.64748952 285 N-acetyl-D-glucosamine M00892 439174 19.66265233 174 Nonanoic acid C01092 8158 10.89687022 215 oleic acid C00712 445639 20.20314755 339 oleic acid 2 C00712 445639 20.25959745 339 palmitic acid C00249 985 18.61732025 313 palmitoleic acid C08362 445638 18.42176534 311 pantothenic acid C00864 6613 18.05143512 291 Phenylalanine C00079 6140 14.17872465 218 phosphoric acid C00009 1004 9.692166033 299 Putrescine C00022 1045 15.4321004 174 pyruvic acid 1060 6.829116202 174 Ribo-5-Hexulosonic acid 630503 15.71482813 217 Ribofuranose C16639 5779 13.95594052 217 ribose C00117 10975657 14.7009099 73 Serine C00065 5951 9.69445116 204 stearic acid C01530 5281 20.43118917 341 stigmasterol C05442 5280794 28.94251938 484 succinic acid C00042 1110 10.25996406 247 threonine C00188 6288 11.17098981 240 trehalose 7427 24.37915148 73 uracil C00106 1174 10.56817642 241 Valine C00015 6029 22.13815678 217 METABOLITES_END #END