#METABOLOMICS WORKBENCH DavideS_20240228_063403 STUDY_ID:ST003113 ANALYSIS_ID:AN005099 #ANALYSIS TYPE: MS; ION MODE: HILIC NEGATIVE ION MODE; (Institute: San Raffaele University) VERSION 1 CREATED_ON 08-05-2024 #PROJECT PR:PROJECT_TITLE Inhibition of Asparagine Synthetase Effectively Retards Polycystic Kidney PR:PROJECT_TITLE Disease Progression. PR:PROJECT_SUMMARY Polycystic Kidney Disease (PKD) is a genetic disorder characterized by bilateral PR:PROJECT_SUMMARY cyst formation. We showed that PKD cells and kidneys display metabolic PR:PROJECT_SUMMARY alterations, including the Warburg effect and glutaminolysis, sustained in vitro PR:PROJECT_SUMMARY by the enzyme asparagine synthetase (ASNS). Here, we used antisense PR:PROJECT_SUMMARY oligonucleotides (ASO) against ASNS in orthologous and slowly progressive PKD PR:PROJECT_SUMMARY murine models and show that treatment leads to a drastic reduction of total PR:PROJECT_SUMMARY kidney volume (measured by MRI) and a prominent rescue of renal function in the PR:PROJECT_SUMMARY mouse. Mechanistically, the upregulation of an ATF4-ASNS axis in PKD is driven PR:PROJECT_SUMMARY by the amino acid response (AAR) branch of the integrated stress response (ISR). PR:PROJECT_SUMMARY Metabolic profiling of PKD control kidneys treated with ASNS-ASOScr-ASO revealed PR:PROJECT_SUMMARY major changes in the mutants, several of which are rescued by ASNS silencing in PR:PROJECT_SUMMARY vivo. Indeed, ASNS drives glutamine-dependent de novo pyrimidine synthesis and PR:PROJECT_SUMMARY proliferation in cystic epithelia. Notably, while several metabolic pathways PR:PROJECT_SUMMARY were completely corrected by ASNS-ASO, glycolysis was only partially restored. PR:PROJECT_SUMMARY Accordingly, combining the glycolytic inhibitor 2DG with ASNS-ASO further PR:PROJECT_SUMMARY improved efficacy. Our studies identify a new therapeutic target and novel PR:PROJECT_SUMMARY metabolic vulnerabilities in PKD. Altogether, targeted metabolomics analysis PR:PROJECT_SUMMARY performed in Tam-Cre;Pkd1ΔC/flox mouse model kidneys corroborates the central PR:PROJECT_SUMMARY role of ASNS in the metabolic rewiring occurring in PKD, highlighting the PR:PROJECT_SUMMARY therapeutic potential of its inhibition. PR:INSTITUTE San Raffaele University PR:LAST_NAME Stefanoni PR:FIRST_NAME Davide PR:ADDRESS Via Olgettina 58, Milan, Milan, 20132, Italy PR:EMAIL stefanoni.davide@hsr.it PR:PHONE +393337686005 PR:DOI http://dx.doi.org/10.21228/M8VM8W #STUDY ST:STUDY_TITLE Inhibition of Asparagine Synthetase Effectively Retards Polycystic Kidney ST:STUDY_TITLE Disease Progression, investigated with targeted tracing metabolomics analysis in ST:STUDY_TITLE MEF cells using 15N2-glutamine. ST:STUDY_SUMMARY Polycystic Kidney Disease (PKD) is a genetic disorder characterized by bilateral ST:STUDY_SUMMARY cyst formation. We showed that PKD cells and kidneys display metabolic ST:STUDY_SUMMARY alterations, including the Warburg effect and glutaminolysis, sustained in vitro ST:STUDY_SUMMARY by the enzyme asparagine synthetase (ASNS). Here, we used antisense ST:STUDY_SUMMARY oligonucleotides (ASO) against Asns in orthologous and slowly progressive PKD ST:STUDY_SUMMARY murine models and show that treatment leads to a drastic reduction of total ST:STUDY_SUMMARY kidney volume (measured by MRI) and a prominent rescue of renal function in the ST:STUDY_SUMMARY mouse. Mechanistically, the upregulation of an ATF4-ASNS axis in PKD is driven ST:STUDY_SUMMARY by the amino acid response (AAR) branch of the integrated stress response (ISR). ST:STUDY_SUMMARY Metabolic profiling of PKD or control kidneys treated with Asns-ASO or Scr-ASO ST:STUDY_SUMMARY revealed major changes in the mutants, several of which are rescued by Asns ST:STUDY_SUMMARY silencing in vivo. Indeed, ASNS drives glutamine-dependent de novo pyrimidine ST:STUDY_SUMMARY synthesis and proliferation in cystic epithelia. Notably, while several ST:STUDY_SUMMARY metabolic pathways were completely corrected by Asns-ASO, glycolysis was only ST:STUDY_SUMMARY partially restored. Accordingly, combining the glycolytic inhibitor 2DG with ST:STUDY_SUMMARY Asns-ASO further improved efficacy. Our studies identify a new therapeutic ST:STUDY_SUMMARY target and novel metabolic vulnerabilities in PKD. Of interest, in these tracing ST:STUDY_SUMMARY studies we could confirm that the pyrimidine biosynthesis pathway is increased ST:STUDY_SUMMARY and rescued by silencing of Asns. ST:INSTITUTE San Raffaele University ST:LAST_NAME Stefanoni ST:FIRST_NAME Davide ST:ADDRESS Via Olgettina 58, Milan, Milan, 20132, Italy ST:EMAIL stefanoni.davide@hsr.it ST:PHONE +393337686005 ST:SUBMIT_DATE 2024-02-28 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - 2DG_1 Sample source:Brain | Treatment:2DG Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=2DG_1.raw SUBJECT_SAMPLE_FACTORS - 2DG_2 Sample source:Brain | Treatment:2DG Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=2DG_2.raw SUBJECT_SAMPLE_FACTORS - 2DG_3 Sample source:Brain | Treatment:2DG Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=2DG_3.raw SUBJECT_SAMPLE_FACTORS - 2DG_4 Sample source:Brain | Treatment:2DG Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=2DG_4.raw SUBJECT_SAMPLE_FACTORS - 2DG_5 Sample source:Brain | Treatment:2DG Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=2DG_5.raw SUBJECT_SAMPLE_FACTORS - Saline_1 Sample source:Brain | Treatment:Saline Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=Saline_1.raw SUBJECT_SAMPLE_FACTORS - Saline_2 Sample source:Brain | Treatment:Saline Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=Saline_2.raw SUBJECT_SAMPLE_FACTORS - Saline_3 Sample source:Brain | Treatment:Saline Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=Saline_3.raw SUBJECT_SAMPLE_FACTORS - Saline_4 Sample source:Brain | Treatment:Saline Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=Saline_4.raw SUBJECT_SAMPLE_FACTORS - Saline_5 Sample source:Brain | Treatment:Saline Sample_Data:Genotype=WT; RAW_FILE_NAME(Raw File Name)=Saline_5.raw #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 #COLLECTION CO:COLLECTION_SUMMARY Animals were euthanized by CO2 inhalation 16 hours following the final injection CO:COLLECTION_SUMMARY and transcardially perfused with ice-cold PBS. Whole brains and spinal cords CO:COLLECTION_SUMMARY were then excised, flash frozen in liquid N2, and stored at –80°C for CO:COLLECTION_SUMMARY biochemical analysis. CO:SAMPLE_TYPE Brain #TREATMENT TR:TREATMENT_SUMMARY Mice were administered i.p. injections of saline (vehicle) or 4 g/kg 2DG (Cayman TR:TREATMENT_SUMMARY Chemical) twice weekly for six weeks. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Metabolites from equal amounts of frontal cortex tissue were rapidly extracted SP:SAMPLEPREP_SUMMARY in 80% ice-cold methanol. Extracted samples were vortexed twice, cleared by SP:SAMPLEPREP_SUMMARY centrifugation at 14,000 x g for 20 minutes at 4°C, and stored at -80°C. #CHROMATOGRAPHY CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME SeQuant ZIC- pHILIC (150 x 2.1mm,5um) CH:COLUMN_TEMPERATURE 30 CH:FLOW_GRADIENT 85% to 30% A in 20 min followed by a wash with 30% A and re-equilibration at 85% CH:FLOW_GRADIENT A CH:FLOW_RATE 150 μL/min CH:SOLVENT_A 100% acetonitrile CH:SOLVENT_B 100% acetonitrile CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_DISPLAY HILIC NEGATIVE ION MODE AN:NUM_FACTORS 2 AN:NUM_METABOLITES 204 AN:UNITS peak intensity #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS 1. A Sequant ZIC-pHILIC column (2.1 mm i.d. × 150 mm, particle size of 5 µm, MS:MS_COMMENTS Millipore Sigma) was used for separation of metabolites. A 2.1 × 20 mm MS:MS_COMMENTS guard column with the same packing material was used for protection of the MS:MS_COMMENTS analytical column. Flow rate was set at 150 μL/min. Buffers consisted of 100% MS:MS_COMMENTS acetonitrile for mobile phase A, and 100% acetonitrile for mobile phase B. The MS:MS_COMMENTS chromatographic gradient ran from 85% to 30% A in 20 min followed by a wash with MS:MS_COMMENTS 30% A and re-equilibration at 85% A. Column temperature was 30 C. MS:ION_MODE NEGATIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS MS_METABOLITE_DATA_START Samples 2DG_1 2DG_2 2DG_3 2DG_4 2DG_5 Saline_1 Saline_2 Saline_3 Saline_4 Saline_5 Factors Sample source:Brain | Treatment:2DG Sample source:Brain | Treatment:2DG Sample source:Brain | Treatment:2DG Sample source:Brain | Treatment:2DG Sample source:Brain | Treatment:2DG Sample source:Brain | Treatment:Saline Sample source:Brain | Treatment:Saline Sample source:Brain | Treatment:Saline Sample source:Brain | Treatment:Saline Sample source:Brain | Treatment:Saline 1-Methyladenosine 18071.7200 29601.5900 20265.8600 20179.1200 25176.2100 20435.6300 15805.1200 26556.5600 18352.0300 27356.2300 1-Methylnicotinamide 2DGP 34911160 33311060 32651360 30735370 33872620 254379.8000 236313.5000 233092 209570.3000 217287.6000 2-Hydroxycaproic acid 252036.5000 336503.3000 150708.8000 182674.3000 208359 342401.1000 268423 231255.7000 224091.6000 303093.6000 2-Hydroxyvaleric acid 2022956 1894132 1561093 1544935 3276415 3049344 2903548 3769964 2819770 2507179 3-Hydroxymethylglutaric acid 13556870 15763370 14454340 13115170 15221390 17466840 15363070 17481170 16075470 15261380 3-Indolepropionic acid 5807.5220 17724.8600 7704.7660 8368.4310 5648.7450 8815.4460 5694.4660 6909.2970 10304.3300 3-Methylhistidine 12789620 9603358 9999853 8921684 8885028 16566540 12191780 16826830 10785260 13711690 3-Phosphoglyceric acid 620132.8000 116324.7000 422951.6000 219568.2000 458680.8000 344162.8000 206785.6000 163721.6000 172609.3000 176084.9000 4-Acetamidobutanoic acid 46045.8100 43092.7700 32575.3400 39037.8900 46626.9100 39254.0800 29085.1300 46967.6600 35428.6300 32975.0900 4-Guanidinobutanoic acid 25369180 35409220 30266640 28637260 25523500 34845760 32768170 19450590 33898070 34336890 4-Hydroxyproline 686297.9000 785089.4000 760941.4000 370256.6000 1042048 582524.2000 590611.4000 754577.1000 640397.9000 684553.6000 4-Pyridoxic acid 40150.0900 34054.9700 20591.1500 20710.5300 33073.8900 27534.2900 32278.2400 56385.5500 24461.6800 32769.6100 4-Trimethylammoniobutanoic acid 44740840 16544860 24587280 20609160 39557420 22627920 19744900 46942940 15877420 18876510 5-Thymidylic acid 21217.0200 20952.5600 13723.1200 6-Phosphogluconic acid 131992.5000 131688.5000 175743.7000 165336.5000 145896 189156.5000 157567.1000 107400.6000 109946 137909.7000 6-Phosphonoglucono-D-lactone Acetylcholine 30827770 10730870 8887630 9589687 14276310 23476840 13938970 31426450 11610480 11394450 Acetyl-CoA 48207.9700 48103.2200 43729.8900 45862.5700 49953.1400 45896.3500 70377.6600 42575.2300 48469.4100 Acetylcysteine 17751.2400 9495.1610 11938.7300 8304.9470 15173.4900 7589.5460 7026.1460 14813.0400 9404.0460 9958.9960 Adenine 57532370 78896990 48441640 51670050 66133490 72162600 72289640 80030930 73680770 77598780 Adenosine 510359200 569644700 378821000 362377200 600005100 420762400 415815200 770467500 471018300 476921600 Adenosine monophosphate 293154700 166545300 251022600 234724100 243260000 339932600 295190100 286926300 238276700 231522900 Adenosine triphosphate 55298.8200 42226.7800 58668.5700 44131.6300 64070.8100 39622.9800 45921.4500 41131.2900 31888.8700 25967.8200 ADP 1741428 541010.4000 1250016 1025581 1670941 957723.4000 1335735 886248.4000 725768.8000 841852.4000 ADP-glucose 48651.9700 57930.4800 50349.3900 52831.9100 47750.5900 89235.5000 70518.1500 73597.6400 62812.5500 62514.8500 AICAR 265721.9000 268141.1000 252398.9000 227734.9000 294959.6000 156866.6000 163426.2000 140513 125417.4000 114242.2000 Allantoin 564283.9000 468667.9000 364734.9000 295323.6000 543052.2000 590014.9000 537383.6000 767742.6000 486362.4000 514008.1000 Aminoadipic acid 1927212 2144710 2343838 1189015 2065791 2434013 2130197 1831133 1602844 1604167 Anserine 898437.7000 719937.8000 911614.5000 990996.8000 744031.3000 1312312 801825.2000 1242324 1102723 1168963 Argininosuccinic acid 848237.1000 1336935 969789.8000 859067.5000 984727.5000 894632 808320.9000 939220.3000 1006648 870770.3000 Asymmetric dimethylarginine 4019863 3466609 2902151 4300072 3969144 4895214 4107970 4765254 3499095 4228862 Betaine 145072900 190533200 179342200 130802300 150686100 172759600 169059300 136329800 160977300 178806700 Butyrylcarnitine 26648530 18788020 19506820 35396130 21041330 28525710 33811250 19440730 22058050 23480590 Caprylic acid 1332729 1554668 1189772 1227729 1232416 1514423 1280417 1086439 1387895 1305085 Carnosine 140248600 145589600 127975500 130758100 159177600 155700300 136089700 233230400 133327400 147243900 CDP 17262.2300 17467.6700 CDP-Ethanolamine 2994532 1807672 1777802 1649207 2560438 2250865 1906025 3020473 1729336 1761456 Choline 28128390 59231190 44411680 35941120 38339280 45109910 32944530 42595220 53113650 42003240 cis-Aconitic acid 2213871 5405462 3586681 4108968 2541130 4870676 3306326 5602864 3980751 3712438 Citicoline 9066455 7897352 6801960 7131539 8017035 9256754 8831307 9786534 6930339 9249086 Citric acid 10026030 21837030 21909960 19151100 12632860 20389100 18771260 28591090 17225750 15299200 Citrulline 7329839 16771450 15785440 17048920 8420502 17723310 18837290 11471360 19635650 16579700 Coenzyme A 95911.1800 93145.9500 Creatine 1833928000 2151483000 2019593000 2012936000 1746102000 2323133000 2112693000 1835404000 1965508000 2035944000 Creatinine 50762710 61545550 50134150 60339260 52575520 66797980 53017040 48757900 49998500 51273970 Cyclic AMP 277023 114447 179289.9000 131296.6000 198919.6000 224148.8000 161820.3000 243043.7000 162981.6000 134656.1000 Cytidine 8217764 2048669 4917061 4313228 8570116 5265363 2637897 9639603 1959524 2700455 Cytidine monophosphate 3923832 3075407 2989602 2621002 3535552 3987320 2648752 5139000 2803018 2570240 Cytidine triphosphate Cytosine 492460.3000 293817.5000 403379.1000 527191.9000 701120.7000 432444.1000 227370.5000 649479.6000 210207.9000 272263.1000 D-2-Hydroxyglutaric acid 7191603 6806436 7724898 6713837 7715719 7276678 6662210 7214574 7221464 6159315 dCDP dCMP 96914.0800 126491.6000 72890.5200 88209.5600 50287.6000 52400.3400 dCTP Deoxyadenosine 153094.6000 114441.6000 184214.7000 144851.7000 230452.8000 135148.1000 86496.0300 235013.3000 100267.8000 115570 Deoxyadenosine monophosphate 96305.1400 78001.0700 72692.1600 61290.5900 112393.4000 122853.6000 89957.7000 131256.7000 85203.1500 84493.0600 Deoxyadenosine triphosphate Deoxycytidine 119932.2000 69858.7000 85630.8400 130680.8000 168254.2000 88129.0200 41421.2200 169985.1000 39148.7800 62967.5000 Deoxyinosine 14943.6800 35460.3800 18412.2500 24016.4900 30338.6000 20728.3600 19289.4900 28597.3500 18580.3800 21904.7300 Deoxyribose 5-phosphate 10633.2300 14462.9400 11770.7900 14751.5900 15482.8500 18280.4200 19039.0200 15805.9500 12488.1900 Deoxyuridine 558577.4000 814494.6000 525856.7000 588276 735457.6000 565671.2000 542528.4000 766922.4000 619638.6000 689229.8000 D-Erythrose 4-phosphate 34583.1600 245135 218971.3000 466760.2000 43650.8900 47358.3000 31110.7100 57304.4500 63008.1400 47446.3800 D-Fructose 134635.6000 189447.7000 259961.1000 223273.5000 127770.9000 125348.7000 103149.6000 71718.4100 125209.4000 151029.8000 D-Glucose 250109.7000 764354.6000 714518.9000 699119 201025.7000 505267.5000 432344.7000 193416.3000 526502.9000 597121 D-Glucuronic acid D-Glyceraldehyde 3-phosphate 223782 1764489 731308.9000 906232.9000 217110.1000 2010465 521459.5000 314252.4000 1078847 583956.6000 D-Ribose 20054.6700 84090.3800 34627.8900 43566.6300 42341.7100 54637.9900 31547.2100 48651.3900 50362.6800 46458.0900 D-Ribose 5-phosphate 1393603 5593067 3919944 4240625 1858809 3399142 1903590 2528462 3188505 2665695 D-Ribulose 9483.2030 9138.3440 9471.7490 7985.9830 13613.0900 5078.5760 7218.3220 7339.0120 7355.7480 D-Ribulose 5-phosphate 4658601 8488487 6325812 6625711 7113898 6367228 6532743 6971767 6518988 7116371 D-Sedoheptulose 7-phosphate 1104049 5169120 2998202 4612077 1447989 2516497 2180161 2663040 3026538 2974881 FAD 109993.2000 173626.6000 82758.8700 141551 112609.4000 107904.7000 163523.5000 183226.7000 117925.1000 116778.8000 Flavin Mononucleotide 25809.7500 36695.7900 26567.7000 41122.0300 23690.3100 62693.3600 32145.1000 26835.5600 38300.4500 31175.8500 "Fructose 1_6-bisphosphate" 79428.4600 200686.9000 352079.8000 450790.4000 92072.3000 166688.8000 80880.0200 35258.0300 147746.2000 105527.1000 Fructose 6-phosphate 1774421 6066211 8259710 10881810 1804326 1769247 1495368 1132382 1930520 1895467 Fumaric acid 11249210 11641450 12071480 11150920 11896950 12150260 11485660 11654570 11844880 10913570 Galactonic acid 2572147 2372503 2132341 2102131 2805436 2632187 2625068 3320245 2154710 2473154 GDP-glucose 90535.5500 85573.7000 68783.3400 105902.8000 120816.5000 Gluconic acid 2199095 2883238 2030624 1728505 1576775 4922440 3529050 3448132 3492978 3015898 Glucosamine Glucosamine 6-phosphate 19895.1200 59251.6800 34858.9900 52045.9900 19477.9600 11439.8700 14762.5000 Glucose 6-phosphate 3059331 14885770 13755350 24007980 3980102 4059717 2437110 1936476 3932645 3790421 Glutathione 164299700 76333340 137238200 120705200 132808200 87940340 108572800 216691500 102424600 135209300 Glycerol 3-phosphate 26184630 43164420 18150170 25744110 25032800 42404910 51261430 31084530 40483270 37934970 Glycerophosphocholine 394422700 347370500 341438600 256990700 368485100 455615600 418268500 361813100 368000200 354391400 Glycine 3198516 7304541 5712069 6225623 3835757 6456195 5830889 3485059 7323194 7181869 Guanidoacetic acid 35710.8600 76919.6300 58362.5700 32101.8100 43280.0700 90652.9400 94967.4500 64143.4100 93570.4300 80199.2000 Guanine 22013.8000 756979.6000 571526.9000 604533.6000 27430.4300 483448.8000 764368.9000 42199.7300 752154.1000 981524.1000 Guanosine 2436505 4563399 1949732 2403832 3685443 2871220 2765883 4861477 2857262 2806479 Guanosine diphosphate 321946.5000 215110.8000 229277.6000 197128.6000 297940.7000 312500.9000 397383.1000 376711.2000 206730.4000 242842.3000 Guanosine monophosphate 19083800 12534680 12810240 11827670 15585410 20760450 16168610 20217990 13464660 14611660 Guanosine triphosphate Hexanoylcarnitine 510087.5000 170820.2000 295258.4000 494415.3000 372963.2000 376565.7000 390465.9000 290666.2000 249012.8000 242017.4000 Hippuric acid 84371.5600 134606.4000 51921.9700 41003.9600 74334.5200 20457 9174.3650 29934.0400 14668.7100 16019.0400 Hypotaurine 168675.2000 76549.9700 141992.5000 134918.2000 167257.9000 131456.8000 119176.9000 259342.5000 105597.3000 104559.6000 Hypoxanthine 53215510 130455000 92819020 108529000 86797430 96021450 100917100 97776440 116058200 116248300 IDP Imidazoleacetic acid 1071584 710556.3000 530864 1031219 816376.9000 851835.2000 566866.2000 725239.8000 587041.4000 490585.2000 Indoxyl sulfate 275574.5000 204074.3000 52635.1800 85993.2300 68886.7300 99807.7000 113313.6000 135910.2000 55563.6100 89933.1300 Inosine 14192020 39707510 20528380 27536150 20652690 28510180 26745320 22841140 29010300 30633180 Inosinic acid 10160090 12773920 10577330 11747270 7902077 22332450 12312560 17243490 16556740 10537980 Isovalerylcarnitine 3351592 3494711 4146262 4996256 3658849 4436381 4015708 2643996 3930333 3762865 Kynurenic acid L-Acetylcarnitine 485208800 576471300 627491600 614630100 470532300 646756100 642480700 467852100 592512600 621154000 L-Alanine 25007680 29212140 27048180 27881570 23571570 32136020 28459590 23653340 27190650 26853370 L-Arginine 60114290 86170920 65192550 70563740 75255760 67489440 81000020 67020880 73745180 77800730 L-Asparagine 18162320 17066960 15019840 14723200 20918040 20074030 17274640 21091820 15422110 16308660 L-Aspartic acid 141107900 175783700 164774900 165413500 155568400 187391500 178792900 161560300 175878000 182614800 L-Carnitine 333972800 415824600 398884400 425403600 345528400 462080400 470982800 329747000 503255300 525087900 L-Cystathionine 6181334 14829700 10590280 18516550 4670692 13199140 14187450 4530979 13334510 14968990 L-Cysteine L-Cystine L-Dihydroorotic acid L-Glutamic acid 1171126000 1036242000 1104980000 1002696000 1163889000 1193687000 1096290000 1240860000 1090286000 1008334000 L-Glutamine 499613700 458360600 473904300 911605500 441539400 534085200 508338100 469651100 470303600 518410100 L-Glyceric acid 1545134 1547527 1407257 1512964 1790321 1629266 1259590 1475182 1269808 1240076 L-Histidine 48880620 75197430 55720250 63582890 53710000 70679860 69921620 66596160 68323180 73814020 L-Homoserine 13974850 14100290 14807880 10766010 15232200 15284830 14675370 16294680 12476300 13908120 L-Isoleucine 10239550 12144630 9347263 10717060 12960180 11478550 11539670 14291570 10210970 11978110 L-Kynurenine 28576.2300 46042.3200 34184.9300 125234.7000 45539.7600 56531.8500 33780.3100 44559.2700 35063.8000 43771.9500 L-Lactic acid 198889500 303803900 239622700 236323800 295438700 313925100 218441400 293174900 286630800 290927900 L-Leucine 25054010 30429620 22375220 26230160 34026400 26968480 27692200 32057840 23933440 32907760 L-Lysine 19511700 19420590 18003250 15512960 22055630 18446430 20026460 20648570 18740880 20473170 L-Methionine 16423710 14160470 15870890 15823760 17553480 15887360 14440890 16881990 14134590 12795470 L-Palmitoylcarnitine 54793.8400 268921.7000 18603.4200 143956.8000 444658.8000 896803.3000 137655.8000 556331.3000 58810.4000 123159.4000 L-Phenylalanine 27331800 31660210 25570030 50035660 33298970 28625300 26820340 30860120 25835580 28979330 L-Pipecolic acid 2724620 4688798 4389446 1600579 2947324 4356211 3989733 2963563 3985155 4193422 L-Proline 68271030 59578980 51282120 61680440 71410970 72071650 54259660 83211280 50576600 51439060 L-Serine 34305820 20371820 24224960 24599180 42576740 25037290 21508940 31725220 22023430 20103120 L-Threonine 10252770 12516080 12095310 9833918 10891850 12919280 12566160 11969030 12104340 12731200 L-Tryptophan 4442551 4972969 4228955 5721814 4695363 5272819 4502810 5071307 4645617 3789705 L-Tyrosine 9999107 9218775 8244602 14952220 10455130 11514560 10816740 12651490 8920834 8466862 L-Valine 18273970 17149250 15858160 16636660 21560880 19135970 17868430 22352720 17602200 17482810 Malic acid 84443200 86589660 95251260 83576870 89325870 91049120 85838140 86616720 88471350 82752640 Myoinositol 71507850 116903100 100772000 65642430 84874860 126661400 122199600 99834620 125234300 118437500 N6-Acetyl-L-lysine 1156396 1115765 1211703 1055036 1244988 1346553 1103837 1413102 1053151 1296254 "N6_N6_N6-Trimethyl-L-lysine" 28493670 8855480 14109950 10964970 29578920 14342100 11652130 34716880 9478067 10778970 N-Acetylasparagine 182604.2000 216333.5000 145093.3000 286098.3000 219083.2000 563391.9000 152606.2000 234285.8000 171803 131359 N-Acetylaspartylglutamic acid 20047000 113241000 76253140 104650300 20297790 113351600 107011500 20399550 104050400 94184090 N-Acetyl-D-glucosamine 69181.9100 52859.0300 43295.2100 41124.5400 59769.5400 25480.0400 28121.9000 40740.1300 26436.4200 28557.6100 N-Acetyl-glucosamine 1-phosphate 597438.8000 444465 382898 311723.6000 682386.7000 528606.5000 391119.1000 934853.3000 353156.1000 419429.3000 N-Acetylglutamic acid 2134306 2326628 2022414 2228409 2301029 2695134 2646070 2607015 2360902 2440022 N-Acetylglutamine 543355.2000 438078 544412.1000 1507291 480796.2000 574506.8000 565452.9000 494582.7000 421792.4000 475130.8000 N-Acetyl-L-alanine 482413.1000 720468.3000 458414 521606.7000 624083.4000 562172.1000 512655.6000 685275 543739.3000 540854.6000 N-Acetyl-L-aspartic acid 802369200 686457200 700385900 664248300 823075200 882483300 722661500 927217800 746456700 745158300 N-Acetyl-L-methionine 1178556 2231410 931591.8000 985970.1000 2053558 1046309 910694.7000 2432937 1189912 1542334 N-Acetylornithine 335052.4000 270672.8000 277713.8000 332240.3000 339471.2000 365257.4000 299814.6000 427717.1000 309466.2000 324432.2000 N-Acetylputrescine N-acetyltryptophan 7467.3990 11443.5200 10140.0500 20234.1200 10269.9900 10137.4500 10598.3700 7235.3240 10592.2800 9261.4870 NAD 8244564 995987.6000 2308486 485484.4000 4224093 3577133 2896259 6366420 1277811 1282631 NADH 1509991 168464.5000 468341.2000 278388.7000 644260.7000 395827.1000 520835.7000 981453.4000 220225.1000 241978.8000 NADP 38173.2500 17435.9700 26180.7500 16006.2900 24665.6700 25818.6600 14992.1200 NADPH N-Formyl-L-methionine Niacinamide 104933100 146834300 141850100 123265300 145615000 128668200 128230000 108009500 147685600 130638000 Nicotinamide riboside 578447.8000 617992.1000 499186.3000 460538.8000 565102.9000 584747.5000 746982.1000 905022.5000 563269.5000 611337.4000 Nicotinamide ribotide 368979.3000 108137.1000 167811.5000 160198 308558.8000 305511.6000 318973.4000 427561.5000 159792.7000 169286.6000 Nicotinic acid 110714.6000 136743.5000 121223.1000 135023.2000 117596.1000 136819.4000 134607.1000 146697.1000 136949.4000 120883.9000 Nicotinic acid adenine dinucleotide Nicotinic acid mononucleotide O-Phosphoethanolamine 104188300 39182130 50681200 46250180 100588000 66069940 55066850 93913060 41008430 40828640 Ornithine Orotic acid 685870.9000 642551.5000 662954.1000 805975.2000 814133.2000 558520.9000 598019.4000 864942 588151.9000 597032.1000 Oxalacetic acid Oxidized glutathione 43019020 37642270 31922020 28100600 39201570 57140880 45602780 34128260 37995920 40541780 Oxoglutaric acid 752072.1000 402941.2000 714907.2000 518279.4000 668160.2000 1709290 543811.8000 1603979 485907.3000 428540.7000 Palmitic acid 4040325 4956063 3818611 4240163 6590868 12192530 4106716 12971930 3515703 3700767 Pantothenic acid 23592810 39073830 32274740 43867970 24600230 33433020 35709090 29699470 34362310 43304320 Phosphocreatine 1512844 903270.2000 680531.4000 799486.7000 584605.8000 1934496 1186722 1682561 1176580 1099153 Phosphoenolpyruvic acid 96672.0600 16353.2300 41097.3500 32119.8300 78038.5200 67605.1900 37022.3900 95052.1800 24773.6400 29923.5900 Phosphorylcholine 463046400 327266700 400295600 413947100 482791800 473420900 421561600 548260500 356150800 393224800 Phosphoserine 30632.1500 21823.9900 27871.4100 20770.7400 42483.6100 21170.2800 19965.7100 33177.6100 23397.4500 19628.6000 Propionylcarnitine 12114170 19382400 14558500 12640900 12996360 29589480 29131320 16970190 20271520 26398620 Purine 20842.2600 14709.5700 25033.4500 15478.2000 6732.9170 9238.5630 8561.9530 11031.7300 10050.4500 Pyridoxine Pyroglutamic acid 9083174 16394620 12944100 21875160 11164020 11149900 9190057 12387660 13133910 13866550 Pyruvic acid 608428.6000 564926.6000 577122.5000 575075.4000 556909.2000 701434.4000 625423.7000 619364.8000 606822.4000 580995.8000 Riboflavin 64464.2800 92790.1700 48556.5200 81277.6200 83398.0400 89883.0100 88099.9500 147591.7000 55286.8000 70361.9300 S-Adenosylhomocysteine 3232633 3748679 3122260 2690293 3684940 3414546 2969820 4656133 2716471 2959100 S-Adenosylmethionine 3737208 4378089 3616028 4509134 3927514 4077517 5276703 3660138 3369053 3221664 "Sedoheptulose 1_7-bisphosphate" 12682 19912.2100 22901.4600 33604.2900 16421.0300 19752.6700 9275.1690 6328.0410 8810.6640 10431.5600 Succinic acid 18363620 16544070 9071933 11855620 17622380 23723230 23775370 22407910 15146440 21011110 Succinyl-CoA Taurine 189176400 131441800 147759000 137948600 191595600 163151900 142471300 204979200 139212800 142121100 Thiamine 922140.4000 747482.4000 631164.1000 579685.5000 892064.9000 723324.6000 784844.7000 1039055 507134.1000 696811.8000 Thiamine monophosphate 701236.7000 1637712 967881.7000 1172770 1012379 1002887 1066737 931165 1387748 1238766 Thymidine 547238.8000 897099.7000 570041.9000 672432.2000 845890.1000 472255 461171.1000 643601.8000 474996.6000 550904.2000 Thymidine 5^-triphosphate Thymine 82562.9700 124651.9000 93549.4800 114468.3000 124063.1000 76158.8200 75138.4000 95598.4500 76460.9600 88909.3800 Uracil 3003195 3593269 2368585 2386399 4082146 2460131 2769332 5158349 2638088 2796553 Ureidopropionic acid 402354.9000 583792.1000 451617.7000 528816.1000 524864.2000 574999.2000 639525.6000 468992.6000 538051.2000 708647.7000 Ureidosuccinic acid 291800.1000 104260 166784.6000 143675.9000 209500.2000 114826.5000 140302.4000 197604 131488.8000 95065.2700 Uric acid 1588201 2470242 1519407 1719608 1679056 2557848 2819014 3509999 2289309 2537028 Uridine 23158180 31243110 26794530 28679470 28036620 26356070 27222640 29968550 27944810 28296540 Uridine 5^-diphosphate 38150.7800 11567.2000 20744.3300 18178.5600 39095.4300 20421.0500 15375.1200 12740.6500 12835.7200 10849 Uridine 5^-monophosphate 6488815 4522133 4939597 4776241 5251102 7314132 5231483 5976845 5178765 6537770 Uridine diphosphate glucose 2303555 1887415 1925247 2252701 1876842 2557450 2228902 2492264 1726329 1702319 Uridine diphosphate glucuronic acid 31603.8200 26857.2400 40438.8100 20943.7000 Uridine diphosphate-N-acetylglucosamine 2358106 1891389 1595269 2203455 3095284 2529683 3555106 2382372 2278980 Uridine triphosphate Urocanic acid 3023881 1840372 1436514 1409767 2665339 1787312 553826.4000 1336565 1161095 1273898 Xanthine 9495254 13985650 8281733 9815354 14119270 9844726 9437991 18084230 10577490 11874360 Xanthosine MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 1-Methyladenosine 1-Methylnicotinamide 2DGP 2-Hydroxycaproic acid 2-Hydroxyvaleric acid 3-Hydroxymethylglutaric acid 3-Indolepropionic acid 3-Methylhistidine 3-Phosphoglyceric acid 4-Acetamidobutanoic acid 4-Guanidinobutanoic acid 4-Hydroxyproline 4-Pyridoxic acid 4-Trimethylammoniobutanoic acid 5-Thymidylic acid 6-Phosphogluconic acid 6-Phosphonoglucono-D-lactone Acetylcholine Acetyl-CoA Acetylcysteine Adenine Adenosine Adenosine monophosphate Adenosine triphosphate ADP ADP-glucose AICAR Allantoin Aminoadipic acid Anserine Argininosuccinic acid Asymmetric dimethylarginine Betaine Butyrylcarnitine Caprylic acid Carnosine CDP CDP-Ethanolamine Choline cis-Aconitic acid Citicoline Citric acid Citrulline Coenzyme A Creatine Creatinine Cyclic AMP Cytidine Cytidine monophosphate Cytidine triphosphate Cytosine D-2-Hydroxyglutaric acid dCDP dCMP dCTP Deoxyadenosine Deoxyadenosine monophosphate Deoxyadenosine triphosphate Deoxycytidine Deoxyinosine Deoxyribose 5-phosphate Deoxyuridine D-Erythrose 4-phosphate D-Fructose D-Glucose D-Glucuronic acid D-Glyceraldehyde 3-phosphate D-Ribose D-Ribose 5-phosphate D-Ribulose D-Ribulose 5-phosphate D-Sedoheptulose 7-phosphate FAD Flavin Mononucleotide "Fructose 1,6-bisphosphate" Fructose 6-phosphate Fumaric acid Galactonic acid GDP-glucose Gluconic acid Glucosamine Glucosamine 6-phosphate Glucose 6-phosphate Glutathione Glycerol 3-phosphate Glycerophosphocholine Glycine Guanidoacetic acid Guanine Guanosine Guanosine diphosphate Guanosine monophosphate Guanosine triphosphate Hexanoylcarnitine Hippuric acid Hypotaurine Hypoxanthine IDP Imidazoleacetic acid Indoxyl sulfate Inosine Inosinic acid Isovalerylcarnitine Kynurenic acid L-Acetylcarnitine L-Alanine L-Arginine L-Asparagine L-Aspartic acid L-Carnitine L-Cystathionine L-Cysteine L-Cystine L-Dihydroorotic acid L-Glutamic acid L-Glutamine L-Glyceric acid L-Histidine L-Homoserine L-Isoleucine L-Kynurenine L-Lactic acid L-Leucine L-Lysine L-Methionine L-Palmitoylcarnitine L-Phenylalanine L-Pipecolic acid L-Proline L-Serine L-Threonine L-Tryptophan L-Tyrosine L-Valine Malic acid Myoinositol N6-Acetyl-L-lysine "N6,N6,N6-Trimethyl-L-lysine" N-Acetylasparagine N-Acetylaspartylglutamic acid N-Acetyl-D-glucosamine N-Acetyl-glucosamine 1-phosphate N-Acetylglutamic acid N-Acetylglutamine N-Acetyl-L-alanine N-Acetyl-L-aspartic acid N-Acetyl-L-methionine N-Acetylornithine N-Acetylputrescine N-acetyltryptophan NAD NADH NADP NADPH N-Formyl-L-methionine Niacinamide Nicotinamide riboside Nicotinamide ribotide Nicotinic acid Nicotinic acid adenine dinucleotide Nicotinic acid mononucleotide O-Phosphoethanolamine Ornithine Orotic acid Oxalacetic acid Oxidized glutathione Oxoglutaric acid Palmitic acid Pantothenic acid Phosphocreatine Phosphoenolpyruvic acid Phosphorylcholine Phosphoserine Propionylcarnitine Purine Pyridoxine Pyroglutamic acid Pyruvic acid Riboflavin S-Adenosylhomocysteine S-Adenosylmethionine "Sedoheptulose 1,7-bisphosphate" Succinic acid Succinyl-CoA Taurine Thiamine Thiamine monophosphate Thymidine Thymidine 5'-triphosphate Thymine Uracil Ureidopropionic acid Ureidosuccinic acid Uric acid Uridine Uridine 5'-diphosphate Uridine 5'-monophosphate Uridine diphosphate glucose Uridine diphosphate glucuronic acid Uridine diphosphate-N-acetylglucosamine Uridine triphosphate Urocanic acid Xanthine Xanthosine METABOLITES_END