#METABOLOMICS WORKBENCH jravilap_20240827_092849 DATATRACK_ID:5151 STUDY_ID:ST003438 ANALYSIS_ID:AN005651 PROJECT_ID:PR002121 VERSION 1 CREATED_ON August 27, 2024, 10:30 am #PROJECT PR:PROJECT_TITLE Genetic screening and metabolomics identify glial adenosine metabolism as a PR:PROJECT_TITLE therapeutic target in Parkinson’s disease PR:PROJECT_TYPE Metabolomic profiling of Parkinson's Disease Drosophila model PR:PROJECT_SUMMARY Parkinson’s disease (PD) is the second most common neurodegenerative disorder PR:PROJECT_SUMMARY and lacks disease-modifying therapies. We developed a Drosophila model for PR:PROJECT_SUMMARY identifying novel glial- based therapeutic targets for PD. Human α-synuclein is PR:PROJECT_SUMMARY expressed in neurons and individual genes are independently knocked down in PR:PROJECT_SUMMARY glia. We performed a forward genetic screen, knocking down the entire Drosophila PR:PROJECT_SUMMARY kinome in glia in α-synuclein expressing flies. Among the top hits were five PR:PROJECT_SUMMARY genes (Ak1, Ak6, Adk1, Adk2, and awd) involved in adenosine metabolism. PR:PROJECT_SUMMARY Knockdown of each gene improved locomotor dysfunction, rescued PR:PROJECT_SUMMARY neurodegeneration, and increased brain adenosine levels. We determined that the PR:PROJECT_SUMMARY mechanism of neuroprotection involves adenosine itself, as opposed to a PR:PROJECT_SUMMARY downstream metabolite. We dove deeper into the mechanism for one gene, Ak1, PR:PROJECT_SUMMARY finding rescue of dopaminergic neuron loss, α-synuclein aggregation, and PR:PROJECT_SUMMARY bioenergetic dysfunction after glial Ak1 knockdown. We performed metabolomics in PR:PROJECT_SUMMARY Drosophila and in human PD patients, allowing us to comprehensively characterize PR:PROJECT_SUMMARY changes in purine metabolism and identify potential biomarkers of dysfunctional PR:PROJECT_SUMMARY adenosine metabolism in people. These experiments support glial adenosine as a PR:PROJECT_SUMMARY novel therapeutic target in PD. PR:INSTITUTE Broad Institute of MIT and Harvard PR:LAST_NAME Avila-Pacheco PR:FIRST_NAME Julian PR:ADDRESS 415 Main Street PR:EMAIL jravilap@broadinstitute.org PR:PHONE (617) 714-1729 PR:CONTRIBUTORS Sodders M, Kumari N, Okorie E, Shen M, Lakhani M, Marathi A, Sarkar S, Scherzer PR:CONTRIBUTORS CR, Clish C, Feany MB, Olsen AL, Bullock K, Pierce K, Dennis C, Jeanfavre D, PR:CONTRIBUTORS Avila-Pacheco, J #STUDY ST:STUDY_TITLE Unbiased genetic screening and metabolomics identifies glial adenosine ST:STUDY_TITLE metabolism as a therapeutic target in Parkinson’s disease ST:STUDY_SUMMARY Parkinson’s disease (PD) is the second most common neurodegenerative disorder ST:STUDY_SUMMARY and lacks disease-modifying therapies. We developed a Drosophila model for ST:STUDY_SUMMARY identifying novel glial- based therapeutic targets for PD. Human α-synuclein is ST:STUDY_SUMMARY expressed in neurons and individual genes are independently knocked down in ST:STUDY_SUMMARY glia. We performed a large forward genetic screen, knocking down the entire ST:STUDY_SUMMARY Drosophila kinome in glia in α-synuclein expressing flies. Among the top hits ST:STUDY_SUMMARY were five genes (Ak1, Ak6, Adk1, Adk2, and awd) involved in adenosine ST:STUDY_SUMMARY metabolism. Knockdown of each gene improved locomotor dysfunction, rescued ST:STUDY_SUMMARY neurodegeneration, and increased brain adenosine levels. We determined that the ST:STUDY_SUMMARY mechanism of neuroprotection involves adenosine itself, as opposed to a ST:STUDY_SUMMARY downstream metabolite. We dove deeper into the mechanism for one gene, Ak1, ST:STUDY_SUMMARY finding rescue of dopaminergic neuron loss, α-synuclein aggregation, and ST:STUDY_SUMMARY bioenergetic dysfunction after glial Ak1 knockdown. We performed metabolomics in ST:STUDY_SUMMARY Drosophila and in human PD patients, allowing us to comprehensively characterize ST:STUDY_SUMMARY changes in purine metabolism and identify potential biomarkers of dysfunctional ST:STUDY_SUMMARY adenosine metabolism in people. These experiments support glial adenosine as a ST:STUDY_SUMMARY novel therapeutic target in PD. ST:INSTITUTE Broad Institute of MIT and Harvard ST:LAST_NAME Avila-Pacheco ST:FIRST_NAME Julian ST:ADDRESS 415 Main Street ST:EMAIL jravilap@broadinstitute.org ST:PHONE (617) 714-1729 #SUBJECT SU:SUBJECT_TYPE Insect SU:SUBJECT_SPECIES Drosophila melanogaster SU:TAXONOMY_ID 7227 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS - 9Q Sample source:Fly heads | Sample_ID:Synuclein RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0001_OLS_FlyPilot_HP-9Q.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0001_OLS_FlyPilot_CP-9Q_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0001_OLS_FlyPilot_CN-9Q.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0001_OLS_FlyPilot_HN_9Q.raw SUBJECT_SAMPLE_FACTORS - 3S Sample source:Fly heads | Sample_ID:Control RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0002_OLS_FlyPilot_HP-3S.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0002_OLS_FlyPilot_CP-3S_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0002_OLS_FlyPilot_CN-3S.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0002_OLS_FlyPilot_HN_3S.raw SUBJECT_SAMPLE_FACTORS - 2S Sample source:Fly heads | Sample_ID:Control RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0003_OLS_FlyPilot_HP-2S.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0003_OLS_FlyPilot_CP-2S_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0003_OLS_FlyPilot_CN-2S.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0003_OLS_FlyPilot_HN_2S.raw SUBJECT_SAMPLE_FACTORS - 6SA Sample source:Fly heads | Sample_ID:Control + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0004_OLS_FlyPilot_HP-6SA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0004_OLS_FlyPilot_CP-6SA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0004_OLS_FlyPilot_CN-6SA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0004_OLS_FlyPilot_HN_6SA.raw SUBJECT_SAMPLE_FACTORS - 8Q Sample source:Fly heads | Sample_ID:Synuclein RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0005_OLS_FlyPilot_HP-8Q.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0005_OLS_FlyPilot_CP-8Q_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0005_OLS_FlyPilot_CN-8Q.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0005_OLS_FlyPilot_HN_8Q.raw SUBJECT_SAMPLE_FACTORS - 5SA Sample source:Fly heads | Sample_ID:Control + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0006_OLS_FlyPilot_HP-5SA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0006_OLS_FlyPilot_CP-5SA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0006_OLS_FlyPilot_CN-5SA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0006_OLS_FlyPilot_HN_5SA.raw SUBJECT_SAMPLE_FACTORS - 10QA Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0007_OLS_FlyPilot_HP-10QA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0007_OLS_FlyPilot_CP-10QA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0007_OLS_FlyPilot_CN-10QA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0007_OLS_FlyPilot_HN_10QA.raw SUBJECT_SAMPLE_FACTORS - 12QA Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0008_OLS_FlyPilot_HP-12QA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0008_OLS_FlyPilot_CP-12QA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0008_OLS_FlyPilot_CN-12QA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0008_OLS_FlyPilot_HN_12QA.raw SUBJECT_SAMPLE_FACTORS - 1S Sample source:Fly heads | Sample_ID:Control RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0009_OLS_FlyPilot_HP-1S.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0009_OLS_FlyPilot_CP-1S_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0009_OLS_FlyPilot_CN-1S.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0009_OLS_FlyPilot_HN_1S.raw SUBJECT_SAMPLE_FACTORS - 4SA Sample source:Fly heads | Sample_ID:Control + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0010_OLS_FlyPilot_HP-4SA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0010_OLS_FlyPilot_CP-4SA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0010_OLS_FlyPilot_CN-4SA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0010_OLS_FlyPilot_HN_4SA.raw SUBJECT_SAMPLE_FACTORS - 11QA Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0011_OLS_FlyPilot_HP-11QA.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0011_OLS_FlyPilot_CP-11QA_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0011_OLS_FlyPilot_CN-11QA.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0011_OLS_FlyPilot_HN_11QA.raw SUBJECT_SAMPLE_FACTORS - 7Q Sample source:Fly heads | Sample_ID:Synuclein RAW_FILE_NAME_HP=22_0131_HP_21_1207_OLS_FlyPilot/B0012_OLS_FlyPilot_HP-7Q.raw; RAW_FILE_NAME_CP=22_0131_CP_21_1207_OLS_FlyPilot/B0012_OLS_FlyPilot_CP-7Q_2.raw; RAW_FILE_NAME_CN=22_0307_CN_21_1207_OLS_FlyPilot/B0012_OLS_FlyPilot_CN-7Q.raw; RAW_FILE_NAME_HN=22_0126_HN_21_1207_OLS_FlyPilot/B0012_OLS_FlyPilot_HN_7Q.raw #COLLECTION CO:COLLECTION_SUMMARY Metabolomics on fly heads was performed using a platform that relies on a CO:COLLECTION_SUMMARY combination of four non-targeted liquid chromatography mass spectrometry (LC-MS) CO:COLLECTION_SUMMARY methods which measure both polar and non-polar metabolites23. n = 3 replicates CO:COLLECTION_SUMMARY per genotype of 25 fly heads each. CO:SAMPLE_TYPE Heads #TREATMENT TR:TREATMENT_SUMMARY NA #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Sample homogenates were generated by homogenizing the fly heads in 100 µL water SP:SAMPLEPREP_SUMMARY using a TissueLyser II (QIAGEN) bead mill set to two 2 min intervals at 20. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE HILIC CH:INSTRUMENT_NAME Shimadzu Nexera X2 CH:COLUMN_NAME Phenomenex Luna NH2 (150 x 2.1mm, 3um) CH:SOLVENT_A 100% water; 20 mM ammonium acetate; 20 mM ammonium hydroxide CH:SOLVENT_B 75% acetonitrile/25% methanol; 10 mM ammonium hydroxide CH:FLOW_GRADIENT The column was eluted with initial conditions of 10% mobile phase A and 90% CH:FLOW_GRADIENT mobile phase B followed by a 10 min linear gradient to 100% mobile phase A. CH:FLOW_RATE 400 µL/min CH:COLUMN_TEMPERATURE 30C #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 NEGATIVE MS:MS_COMMENTS Raw data were processed using TraceFinder 3.3 software (Thermo Fisher MS:MS_COMMENTS Scientific; Waltham, MA) and Progenesis QI (Nonlinear Dynamics; Newcastle upon MS:MS_COMMENTS Tyne, UK). Metabolite identities were confirmed using authentic reference MS:MS_COMMENTS standards or reference samples. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Abundances MS_METABOLITE_DATA_START Samples 1S 2S 3S 4SA 5SA 6SA 7Q 8Q 9Q 10QA 11QA 12QA Factors Sample source:Fly heads | Sample_ID:Control Sample source:Fly heads | Sample_ID:Control Sample source:Fly heads | Sample_ID:Control Sample source:Fly heads | Sample_ID:Control + AKI-KD Sample source:Fly heads | Sample_ID:Control + AKI-KD Sample source:Fly heads | Sample_ID:Control + AKI-KD Sample source:Fly heads | Sample_ID:Synuclein Sample source:Fly heads | Sample_ID:Synuclein Sample source:Fly heads | Sample_ID:Synuclein Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD Sample source:Fly heads | Sample_ID:Synuclein + AKI-KD Thymine-d4 [iSTD] 229361860 231289575 231273992 231996982 250012415 249150437 240123510 252747906 231624478 224400702 234234518 230246788 Inosine-15N4 [iSTD] 38614413 39111453 41368203 37335176 39156917 38732381 41692619 42426810 39482680 34401891 38426333 36002298 Glycocholate-d4 [iSTD] 96054921 94765270 93122230 91149777 104348515 102905129 92329613 104487560 97725723 95165013 94869811 97491874 1-Methylhistidine 212347 271194 397948 474191 280459 394867 267610 823868 591625 541506 1-Methylxanthine 842352 1137250 948291 1409282 2452019 2056295 1146840 1957015 1901509 3014029 2382446 3721509 2-Hydroxybutyric acid or 3-Hydroxybutyric acid 500460 369986 533357 399793 520645 441213 461873 355645 402107 555958 523080 512082 2-Hydroxyglutaric acid 150564132 124965567 117565017 189492401 174540865 189834603 249174648 250870158 280415238 260492382 269526837 317170401 3-Methyl-2-oxovaleric acid or Ketoleucine 6914807 6066898 8162793 4950901 7211176 6671145 5554374 6320598 6005251 7226425 7617664 6720737 3-Methyladipic acid or Pimelic acid 2085725 1941588 2671000 1481271 2892676 2257301 1205124 2136306 1904163 3469760 2451108 1959485 3-Phosphoglyceric acid 4130319 2640584 2775263 4659312 3724643 3128839 3554020 3120658 3063701 5509889 3333486 6259628 4-Pyridoxic acid 183774 92430 259787 233526 187457 146821 377796 251747 261458 222148 282048 6-Phosphogluconic acid 85760 148952 87136 91322 Adenine 13554341 16589514 14988045 19437685 20276891 22279605 13233503 14109013 12778593 22480761 14741580 13046441 Adenylsuccinic acid 8208667 7068581 5629240 7624708 5673259 7917161 4917112 6010949 6323877 8485395 7204340 6967636 Adipic acid or Methylglutaric acid 2163227 2129357 1692940 3769307 3864862 4266222 2097666 4776729 2167210 7993486 4139331 5803861 Adonitol or Arabitol 66408549 43915200 39120418 97624580 103898306 98146867 45457950 70114862 54694905 74491205 68807784 105043022 ADP 2192225 1928835 2253699 1980469 2067582 2191467 1932924 2187650 2261210 3487154 2216228 2106230 Aminoadipic acid 227178 509891 201369 2187654 225430 216282 1391790 640356 AMP 179488077 146712154 135004982 185468232 153623464 173986323 158186119 151141738 157961623 256726533 183355214 181016765 Aspartic acid 36944435 51592048 52043396 18939431 14822994 25824297 33855312 8134565 23400977 7371019 17778659 11820024 ATP 78217 172582 141361 105734 224134 240728 89951 154080 67922 224367 94112 146880 Citric acid or Isocitric acid 372295784 238872230 291514061 219817444 213776362 246493708 287846794 293691663 334118764 322534415 254283043 239508549 CMP 490033 525733 364111 428025 412808 422131 381563 418398 717694 565418 413238 467450 Cystathionine 824452 457260 577568 908043 498439 490697 435556 284124 379788 805826 725425 653164 Cytidine 445285 647577 533805 706372 856710 905526 585513 531432 568009 696390 725755 859456 Erythronic acid or Threonic acid 50286856 58229106 58849484 41713630 69105441 67710132 51807326 87494777 59474643 41856452 39391330 43447698 GDP 92274 122046 93701 65730 96142 120467 87500 95125 123740 36525 76063 43344 Glucosan or 3-Hydroxymethylglutaric acid 7126144 7432129 7499000 6298558 9013162 8860053 4567787 6974946 6406712 8769058 7683397 7423995 Glucuronic acid 9723052 9408824 9394114 7186856 7528498 8268365 7449608 8182685 8108635 7122016 5867368 7186244 Glutamic acid 189519215 168471987 167643500 214553752 237664912 237093653 157511690 164513289 181010181 169209880 194353286 207011660 Glutathione 12892505 10361158 2598644 33391564 7969737 8388074 26543303 7689410 5754785 32244700 28165815 17866210 Glyceric acid 7075967 6516995 6559614 8622308 8268233 9763859 7091873 12476149 6454200 15107630 7389254 10916451 Glycocholic acid 683654 548127 491976 617726 655532 564361 685043 680042 610922 688538 644249 674725 GMP 29941174 22936864 21504280 28250736 23000591 25482758 26306667 22297255 27380932 30564146 28111514 28958703 Guanosine 80699170 72877119 64108209 70400720 69018763 67861551 74158345 76869386 87081715 79874185 86656255 102215745 Hexose 19000348 18072595 15930476 19757564 24297977 27449295 16929205 25683143 20041670 21912415 18588947 21818038 Hexose diphosphate 619807 597040 445345 493168 529027 594399 528910 758760 686199 659807 599032 670169 Hexose monophosphate 52961510 38774520 32950944 48095071 31203526 33859108 37029017 46863874 37028164 54148492 27952057 53381915 Homovanillic acid 468918 326027 476302 285427 325493 376236 401519 188899 368083 173298 379670 234756 Hydroxykynurenine 1615682 1319023 1418895 4221167 3121177 3631085 3185571 2358294 3225559 4133104 4809121 4402948 Hypoxanthine 16261310 15793099 15337842 18278910 22763622 23266463 15202064 24074265 17376699 27669962 14934572 18531328 IMP 12040803 9270262 7595849 14773495 12250893 13395900 11448424 13578809 12588075 20377252 14440366 15314838 Indolelactic acid 1403193 1219571 1218826 1184247 1099848 1486468 861533 933567 1256826 1227936 1289335 1376493 Inosine 100611427 92496365 83897912 98449868 109502235 107637944 93037390 129045226 101647996 176474140 91078896 118087061 Lactic acid 48996379 46412511 48894055 65969909 55871434 68669494 107766490 102854434 122323500 116873869 93228900 106023286 Malic acid 1024826917 982437304 902235804 982736934 982921037 1051988789 881147641 894684503 886091093 1047229085 910148182 888683787 Malondialdehyde 5148703 4571030 3491233 5437770 4205844 4546308 4599883 5172716 8250558 6530963 5260865 4866232 Malonic acid 714187 871291 884224 565954 908101 811935 576267 781113 651723 787234 775063 743942 Myo-inositol 55715708 52187964 48269478 66711023 76942314 75443488 55261934 62978900 60455813 74279288 55675387 63823072 N-Acetylglutamic acid 59721305 51466875 48279186 68722967 57058227 64921580 84017243 80606885 74057397 109319384 101795310 107957346 NAD+ 312557 661996 314823 399583 765328 622131 189508 451397 337634 597814 276551 372111 Nicotinic acid 1409812 1051387 1102628 1889990 1826716 1965467 1267935 1415218 1338477 1872162 1490010 1862672 Orotic acid 1325826 991036 866256 1145314 1152068 1063636 847607 875808 1001790 1054323 1149843 1047606 Oxalic acid 6036762 4767117 5216804 5701167 4712142 4778957 6232334 4575016 4913227 4540354 4780375 4610007 Oxidized glutathione 722041 1073652 1223659 714795 464837 581784 826239 273581 1348268 59410 602619 132416 Oxoglutaric acid 14443450 7118117 15320202 18534551 18146360 10916533 12208509 21124947 18816424 27649570 21083222 40051240 Pantothenic acid 102109646 133869134 99166604 101618874 120621412 121753299 111945973 124613526 96803328 124312594 107588850 63053972 Pentose 5-phosphate 1508209 1118031 1334983 1788519 1016148 1018462 1243894 1288452 1200750 1244335 937006 1336568 Pseudouridine 12378921 9308109 9740397 21593428 21891302 26414441 10930335 11709924 10981864 13685282 12351927 16474670 Ranitidine 1746171 1760278 2292506 1317603 2411581 2024113 1261983 1910107 1926029 2635006 1966146 1801152 Sebacic acid 748881 818828 956478 779766 1190898 1026624 850948 1013786 1002613 1179787 1553690 1612847 sn-Glycero-3-phosphate 143752131 117329219 106189610 178829691 141680671 135996048 131175838 106974299 128572834 170102064 114033892 131338191 Sorbitol 26915100 31489555 30099340 39748717 38664349 47260134 32117374 40033420 29301876 41885822 25514445 30475319 Suberic acid 1069492 1340876 1505951 1168118 1845622 1544762 1151485 1402998 1422740 1322124 1496093 1714229 Succinic acid 179565919 204446041 145370578 188061030 181055068 167590403 173194735 171583676 186430243 143109387 179987087 177216370 Succinyladenosine 1112131 742248 866080 1773284 876376 1626657 864653 894846 1235004 18925698 801339 2948114 Sucrose or Lactose or Trehalose 96713647 69345773 103537204 70603793 73879202 86779814 125375281 291361256 244970397 46373066 70957259 77245310 Thymine 1763642 1428679 1946692 3163026 3193761 3881598 1199481 1861695 1501226 3075632 2494343 2936656 UMP 6354306 5137027 3789468 6556773 4949012 5668450 4027852 4503148 5820990 6383859 4364013 3501201 Uracil 62028558 60548873 52692604 71817119 83093935 83900119 56903316 71111821 65747906 78290671 62738491 73454658 Uric acid 4388671494 3870934668 3864806080 4400756130 3696981490 4021526030 4028467517 3541472533 3854565656 4514005475 4411703775 4439716809 Uridine 59667583 58692859 45627747 68156536 72111547 73372289 44629780 56184872 52470384 69537995 51397341 60359154 Xanthine 145047271 147719639 197690663 215735647 246022363 275340539 132024254 157497644 155673463 153525396 150739179 169872912 Xanthurenic acid 1065584239 935612799 921653015 1482236550 1336978034 1394192513 1051701104 1027907319 1083493502 1307743925 1310214026 1534088961 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name MZ RT HMDB_ID HMDB_ID_specificity (1=match,2=representative) Thymine-d4 [iSTD] 129.0608 1.29 internal standard NA Inosine-15N4 [iSTD] 271.0616 3.16 internal standard NA Glycocholate-d4 [iSTD] 468.3269 3.63 internal standard NA 1-Methylhistidine 168.0779 3.22 HMDB0000001 1 1-Methylxanthine 165.0418 3.58 HMDB0010738 1 2-Hydroxybutyric acid or 3-Hydroxybutyric acid 103.0401 3.37 HMDB0000008 2 2-Hydroxyglutaric acid 147.0299 6.43 HMDB0000694 1 3-Methyl-2-oxovaleric acid or Ketoleucine 129.0557 3.34 HMDB0000491 1 3-Methyladipic acid or Pimelic acid 159.0663 6.33 HMDB0000555 2 3-Phosphoglyceric acid 184.9857 8.62 HMDB0000807 1 4-Pyridoxic acid 182.0459 3.32 HMDB0000017 1 6-Phosphogluconic acid 275.0174 8.26 HMDB0001316 1 Adenine 134.0472 2.17 HMDB0000034 1 Adenylsuccinic acid 462.0668 9.64 HMDB0000536 1 Adipic acid or Methylglutaric acid 145.0506 6.35 HMDB0000448 2 Adonitol or Arabitol 151.0612 1.71 HMDB0000508 1 ADP 426.0221 9.14 HMDB0001341 1 Aminoadipic acid 160.0615 5.03 HMDB0000510 1 AMP 346.0558 7.23 HMDB0000045 1 Aspartic acid 132.0302 4.93 HMDB0000191 1 ATP 505.9885 10.57 HMDB0000538 1 Citric acid or Isocitric acid 191.0197 8.37 HMDB0000094 2 CMP 322.0446 7.09 HMDB0000095 1 Cystathionine 221.0601 5.17 HMDB0000099 1 Cytidine 242.0782 2.07 HMDB0000089 1 Erythronic acid or Threonic acid 135.0299 4.06 HMDB0000613 2 GDP 442.017 9.87 HMDB0001201 1 Glucosan or 3-Hydroxymethylglutaric acid 161.0455 6.35 HMDB0000640 2 Glucuronic acid 193.0354 4.81 HMDB0000127 1 Glutamic acid 146.0459 4.9 HMDB0000148 1 Glutathione 306.0765 5.59 HMDB0000125 1 Glyceric acid 105.0193 3.95 HMDB0000139 1 Glycocholic acid 464.3018 3.63 HMDB0000138 1 GMP 362.0507 8 HMDB0001397 1 Guanosine 282.0844 2.91 HMDB0000133 1 Hexose 179.0561 2.07 HMDB0000122 2 Hexose diphosphate 338.9888 9.44 HMDB0001058 2 Hexose monophosphate 259.0224 7.04 HMDB0000124 2 Homovanillic acid 181.0506 3.99 HMDB0000118 1 Hydroxykynurenine 223.0724 3.79 HMDB0000732 1 Hypoxanthine 135.0312 3.14 HMDB0000157 1 IMP 347.0398 8.2 HMDB0000175 1 Indolelactic acid 204.0666 3.97 HMDB0000671 1 Inosine 267.0735 3.18 HMDB0000195 1 Lactic acid 89.0244 3.59 HMDB0000190 1 Malic acid 133.0143 6.55 HMDB0000156 1 Malondialdehyde 71.0139 4.31 HMDB0006112 1 Malonic acid 103.0037 6.72 HMDB0000691 1 Myo-inositol 179.0561 2.58 HMDB0000211 1 N-Acetylglutamic acid 188.0565 6.27 HMDB0001138 1 NAD+ 662.1024 4.87 HMDB0000902 1 Nicotinic acid 122.0247 3.7 HMDB0001488 1 Orotic acid 155.0098 4.09 HMDB0000226 1 Oxalic acid 88.988 6.98 HMDB0002329 1 Oxidized glutathione 611.1447 7.39 HMDB0003337 1 Oxoglutaric acid 145.0143 6.88 HMDB0000208 1 Pantothenic acid 218.1034 3.67 HMDB0000210 1 Pentose 5-phosphate 229.0119 6.89 HMDB0001548 1 Pseudouridine 243.0623 2.26 HMDB0000767 1 Ranitidine 313.134 1.07 HMDB0001930 1 Sebacic acid 201.1132 6.26 HMDB0000792 1 sn-Glycero-3-phosphate 171.0064 6.88 HMDB0000126 1 Sorbitol 181.0718 1.93 HMDB0000247 1 Suberic acid 173.0819 6.32 HMDB0000893 1 Succinic acid 117.0193 6.49 HMDB0000254 1 Succinyladenosine 382.1004 6.88 HMDB0000912 1 Sucrose or Lactose or Trehalose 341.1089 2.58 HMDB0000258 2 Thymine 125.0357 1.28 HMDB0000262 1 UMP 323.0286 7.15 HMDB0000288 1 Uracil 111.02 1.51 HMDB0000300 1 Uric acid 167.0211 4.84 HMDB0000289 1 Uridine 243.0623 1.7 HMDB0000296 1 Xanthine 151.0262 4.61 HMDB0000292 1 Xanthurenic acid 204.0302 7.59 HMDB0000881 1 METABOLITES_END #END