#METABOLOMICS WORKBENCH juliehaines_20230221_104003 DATATRACK_ID:3754 STUDY_ID:ST002488 ANALYSIS_ID:AN004062 PROJECT_ID:PR001607 VERSION 1 CREATED_ON February 21, 2023, 11:50 am #PROJECT PR:PROJECT_TITLE Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit PR:PROJECT_TITLE pancreatic cancer growth and metastasis PR:PROJECT_SUMMARY In pancreatic ductal adenocarcinoma (PDAC) patients, we show that response to PR:PROJECT_SUMMARY radiation therapy (RT) is characterized by increased IL2Rβ and IL2Rγ and PR:PROJECT_SUMMARY decreased ILR2α expression. The bispecific aPD1-IL2v is a PD-1-targeted IL-2 PR:PROJECT_SUMMARY variant (IL2v) immunocytokine with engineered IL-2 cis-targeted to PD-1 and PR:PROJECT_SUMMARY abolished IL2Rα binding, which enhances tumor-antigen specific T cell PR:PROJECT_SUMMARY activation while reducing regulatory T cell (Treg) suppression. Using aPD1-IL2v PR:PROJECT_SUMMARY in orthotopic PDAC KPC-driven tumor models, we show marked improvement in local PR:PROJECT_SUMMARY and metastatic survival along with profound increase in tumor-infiltrating PR:PROJECT_SUMMARY polyfunctional CD8+ T cell subsets with a transcriptionally and metabolically PR:PROJECT_SUMMARY active phenotype, and preferential activation of antigen-specific CD8+ T cells. PR:PROJECT_SUMMARY In combination with single dose RT, aPD1-IL2v treatment results in a robust, PR:PROJECT_SUMMARY durable expansion of polyfunctional CD8+ T cells, T cell stemness, PR:PROJECT_SUMMARY tumor-specific memory immune response, natural killer (NK) cell activation, and PR:PROJECT_SUMMARY decreased Tregs. These data show that aPD1-IL2v leads to profound local and PR:PROJECT_SUMMARY distant response in PDAC. PR:INSTITUTE University of Colorado Denver PR:LABORATORY Lab of Angelo D'Alessandro in collaboration with lab of Sana Karam PR:LAST_NAME Haines PR:FIRST_NAME Julie PR:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA PR:EMAIL julie.haines@cuanschutz.edu PR:PHONE 3037243339 #STUDY ST:STUDY_TITLE Simultaneous targeting of PD-1 and IL2Rβγ with radiation therapy to inhibit ST:STUDY_TITLE pancreatic cancer growth and metastasis - mouse serum metabolomics ST:STUDY_SUMMARY C57BL/6 mice were orthotopically implanted with PK5L1940 cells. 8 Gy RT was ST:STUDY_SUMMARY administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week ST:STUDY_SUMMARY beginning day 7 post-implantation. Spleens and lymph nodes were harvested 17 ST:STUDY_SUMMARY days post implantation and the sorted T-cells cultured in the presence of 13C6 ST:STUDY_SUMMARY glucose for 6h. This study contains longitudinal sampling of the cell culture ST:STUDY_SUMMARY supernatants. ST:INSTITUTE University of Colorado Denver ST:LAST_NAME Haines ST:FIRST_NAME Julie ST:ADDRESS 12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA ST:EMAIL julie.haines@cuanschutz.edu ST:PHONE 3037243339 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus #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 - AG-23-TS-1 Factor:Untrx | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-1_r16- SUBJECT_SAMPLE_FACTORS - AG-23-TS-2 Factor:Untrx | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-2_r23- SUBJECT_SAMPLE_FACTORS - AG-23-TS-3 Factor:Untrx | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-3_r48- SUBJECT_SAMPLE_FACTORS - AG-23-TS-4 Factor:RT | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-4_r49- SUBJECT_SAMPLE_FACTORS - AG-23-TS-5 Factor:RT | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-5_r28- SUBJECT_SAMPLE_FACTORS - AG-23-TS-6 Factor:RT | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-6_r4- SUBJECT_SAMPLE_FACTORS - AG-23-TS-7 Factor:IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-7_r53- SUBJECT_SAMPLE_FACTORS - AG-23-TS-8 Factor:IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-8_r20- SUBJECT_SAMPLE_FACTORS - AG-23-TS-9 Factor:IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-9_r35- SUBJECT_SAMPLE_FACTORS - AG-23-TS-10 Factor:RT+IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-10_r41- SUBJECT_SAMPLE_FACTORS - AG-23-TS-11 Factor:RT+IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-11_r46- SUBJECT_SAMPLE_FACTORS - AG-23-TS-12 Factor:RT+IL2v | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-12_r36- SUBJECT_SAMPLE_FACTORS - AG-23-TS-13 Factor:aCD25 | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-13_r14- SUBJECT_SAMPLE_FACTORS - AG-23-TS-14 Factor:aCD25 | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-14_r10- SUBJECT_SAMPLE_FACTORS - AG-23-TS-15 Factor:aCD25 | Timepoint:5 min RAW_FILE_NAME=AG-23-TS-15_r40- SUBJECT_SAMPLE_FACTORS - AG-23-TS-16 Factor:Untrx | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-16_r27- SUBJECT_SAMPLE_FACTORS - AG-23-TS-17 Factor:Untrx | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-17_r31- SUBJECT_SAMPLE_FACTORS - AG-23-TS-18 Factor:Untrx | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-18_r38- SUBJECT_SAMPLE_FACTORS - AG-23-TS-19 Factor:RT | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-19_r54- SUBJECT_SAMPLE_FACTORS - AG-23-TS-20 Factor:RT | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-20_r39- SUBJECT_SAMPLE_FACTORS - AG-23-TS-21 Factor:RT | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-21_r19- SUBJECT_SAMPLE_FACTORS - AG-23-TS-22 Factor:IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-22_r1- SUBJECT_SAMPLE_FACTORS - AG-23-TS-23 Factor:IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-23_r18- SUBJECT_SAMPLE_FACTORS - AG-23-TS-24 Factor:IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-24_r58- SUBJECT_SAMPLE_FACTORS - AG-23-TS-25 Factor:RT+IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-25_r60- SUBJECT_SAMPLE_FACTORS - AG-23-TS-26 Factor:RT+IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-26_r55- SUBJECT_SAMPLE_FACTORS - AG-23-TS-27 Factor:RT+IL2v | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-27_r2- SUBJECT_SAMPLE_FACTORS - AG-23-TS-28 Factor:aCD25 | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-28_r17- SUBJECT_SAMPLE_FACTORS - AG-23-TS-29 Factor:aCD25 | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-29_r22- SUBJECT_SAMPLE_FACTORS - AG-23-TS-30 Factor:aCD25 | Timepoint:1 hr RAW_FILE_NAME=AG-23-TS-30_r6- SUBJECT_SAMPLE_FACTORS - AG-23-TS-31 Factor:Untrx | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-31_r37- SUBJECT_SAMPLE_FACTORS - AG-23-TS-32 Factor:Untrx | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-32_r3- SUBJECT_SAMPLE_FACTORS - AG-23-TS-33 Factor:Untrx | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-33_r24- SUBJECT_SAMPLE_FACTORS - AG-23-TS-34 Factor:RT | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-34_r44- SUBJECT_SAMPLE_FACTORS - AG-23-TS-35 Factor:RT | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-35_r42- SUBJECT_SAMPLE_FACTORS - AG-23-TS-36 Factor:RT | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-36_r26- SUBJECT_SAMPLE_FACTORS - AG-23-TS-37 Factor:IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-37_r50- SUBJECT_SAMPLE_FACTORS - AG-23-TS-38 Factor:IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-38_r13- SUBJECT_SAMPLE_FACTORS - AG-23-TS-39 Factor:IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-39_r12- SUBJECT_SAMPLE_FACTORS - AG-23-TS-40 Factor:RT+IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-40_r29- SUBJECT_SAMPLE_FACTORS - AG-23-TS-41 Factor:RT+IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-41_r51- SUBJECT_SAMPLE_FACTORS - AG-23-TS-42 Factor:RT+IL2v | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-42_r59- SUBJECT_SAMPLE_FACTORS - AG-23-TS-43 Factor:aCD25 | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-43_r52- SUBJECT_SAMPLE_FACTORS - AG-23-TS-44 Factor:aCD25 | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-44_r7- SUBJECT_SAMPLE_FACTORS - AG-23-TS-45 Factor:aCD25 | Timepoint:2 hr RAW_FILE_NAME=AG-23-TS-45_r45- SUBJECT_SAMPLE_FACTORS - AG-23-TS-46 Factor:Untrx | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-46_r43- SUBJECT_SAMPLE_FACTORS - AG-23-TS-47 Factor:Untrx | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-47_r57- SUBJECT_SAMPLE_FACTORS - AG-23-TS-48 Factor:Untrx | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-48_r25- SUBJECT_SAMPLE_FACTORS - AG-23-TS-49 Factor:RT | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-49_r5- SUBJECT_SAMPLE_FACTORS - AG-23-TS-50 Factor:RT | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-50_r8- SUBJECT_SAMPLE_FACTORS - AG-23-TS-51 Factor:RT | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-51_r34- SUBJECT_SAMPLE_FACTORS - AG-23-TS-52 Factor:IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-52_r47- SUBJECT_SAMPLE_FACTORS - AG-23-TS-53 Factor:IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-53_r30- SUBJECT_SAMPLE_FACTORS - AG-23-TS-54 Factor:IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-54_r56- SUBJECT_SAMPLE_FACTORS - AG-23-TS-55 Factor:RT+IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-55_r15- SUBJECT_SAMPLE_FACTORS - AG-23-TS-56 Factor:RT+IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-56_r9- SUBJECT_SAMPLE_FACTORS - AG-23-TS-57 Factor:RT+IL2v | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-57_r33- SUBJECT_SAMPLE_FACTORS - AG-23-TS-58 Factor:aCD25 | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-58_r11- SUBJECT_SAMPLE_FACTORS - AG-23-TS-59 Factor:aCD25 | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-59_r21- SUBJECT_SAMPLE_FACTORS - AG-23-TS-60 Factor:aCD25 | Timepoint:6 hr RAW_FILE_NAME=AG-23-TS-60_r32- #COLLECTION CO:COLLECTION_SUMMARY C57BL/6 mice were orthotopically implanted with PK5L1940 cells. 8 Gy RT was CO:COLLECTION_SUMMARY administered 7 days post-implantation. PD1-IL2v and aCD25 dosed once per week CO:COLLECTION_SUMMARY beginning day 7 post-implantation. Spleens and lymph nodes were harvested 17 CO:COLLECTION_SUMMARY days post implantation and homogenized to a single cell suspension by mashing CO:COLLECTION_SUMMARY through a 70 uM cell strainer. For spleen processing, the erythrocytes were CO:COLLECTION_SUMMARY lysed with ACK (ammonium-chloride-potassium) lysis buffer for 3 min at RT. CD8 T CO:COLLECTION_SUMMARY cells were sorted with a CD8-negative selection Stemcell isolation kit following CO:COLLECTION_SUMMARY manufacturer instructions. CO:SAMPLE_TYPE Cell culture supernatant #TREATMENT TR:TREATMENT_SUMMARY CD8+ T cells were cultured for 6h in the presence of 25 mM U-13C-glucose in TR:TREATMENT_SUMMARY glucose-free RPMI. Media aliquots were taken at 5 min, 1h, 2h, 6h and snap TR:TREATMENT_SUMMARY frozen. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Supernatant aliquots were thawed on ice then a 20 uL aliquot treated with 480 uL SP:SAMPLEPREP_SUMMARY of cold 5:3:2 MeOH:acetonitrile:water. Samples were vortexed 30 min at 4 degrees SP:SAMPLEPREP_SUMMARY C then supernatants clarified by centrifugation (10 min, 10,000 g, 4 degrees C) SP:SAMPLEPREP_SUMMARY and transferred to autosampler vials. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE -80℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Negative C18 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Thermo Vanquish CH:COLUMN_NAME Phenomenex Kinetex C18 2.1 x 150 mm, 1.7 um CH:SOLVENT_A 95% water 5% acetonitrile 1 mM ammonium acetate CH:SOLVENT_B 95% acetonitrile 5% water 1 mM ammonium acetate CH:FLOW_GRADIENT 0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min CH:FLOW_GRADIENT 100-0% B, 3-5 min hold at 0% B CH:FLOW_RATE 450 uL/min CH:COLUMN_TEMPERATURE 45 CH:SAMPLE_INJECTION 10 uL #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, MS:MS_COMMENTS microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, MS:MS_COMMENTS capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas MS:MS_COMMENTS 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were MS:MS_COMMENTS annotated and integrated using Maven in conjunction with the KEGG database. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS peak area MS_METABOLITE_DATA_START Samples AG-23-TS-1 AG-23-TS-2 AG-23-TS-3 AG-23-TS-4 AG-23-TS-5 AG-23-TS-6 AG-23-TS-7 AG-23-TS-8 AG-23-TS-9 AG-23-TS-10 AG-23-TS-11 AG-23-TS-12 AG-23-TS-13 AG-23-TS-14 AG-23-TS-15 AG-23-TS-16 AG-23-TS-17 AG-23-TS-18 AG-23-TS-19 AG-23-TS-20 AG-23-TS-21 AG-23-TS-22 AG-23-TS-23 AG-23-TS-24 AG-23-TS-25 AG-23-TS-26 AG-23-TS-27 AG-23-TS-28 AG-23-TS-29 AG-23-TS-30 AG-23-TS-31 AG-23-TS-32 AG-23-TS-33 AG-23-TS-34 AG-23-TS-35 AG-23-TS-36 AG-23-TS-37 AG-23-TS-38 AG-23-TS-39 AG-23-TS-40 AG-23-TS-41 AG-23-TS-42 AG-23-TS-43 AG-23-TS-44 AG-23-TS-45 AG-23-TS-46 AG-23-TS-47 AG-23-TS-48 AG-23-TS-49 AG-23-TS-50 AG-23-TS-51 AG-23-TS-52 AG-23-TS-53 AG-23-TS-54 AG-23-TS-55 AG-23-TS-56 AG-23-TS-57 AG-23-TS-58 AG-23-TS-59 AG-23-TS-60 Factors Factor:Untrx | Timepoint:5 min Factor:Untrx | Timepoint:5 min Factor:Untrx | Timepoint:5 min Factor:RT | Timepoint:5 min Factor:RT | Timepoint:5 min Factor:RT | Timepoint:5 min Factor:IL2v | Timepoint:5 min Factor:IL2v | Timepoint:5 min Factor:IL2v | Timepoint:5 min Factor:RT+IL2v | Timepoint:5 min Factor:RT+IL2v | Timepoint:5 min Factor:RT+IL2v | Timepoint:5 min Factor:aCD25 | Timepoint:5 min Factor:aCD25 | Timepoint:5 min Factor:aCD25 | Timepoint:5 min Factor:Untrx | Timepoint:1 hr Factor:Untrx | Timepoint:1 hr Factor:Untrx | Timepoint:1 hr Factor:RT | Timepoint:1 hr Factor:RT | Timepoint:1 hr Factor:RT | Timepoint:1 hr Factor:IL2v | Timepoint:1 hr Factor:IL2v | Timepoint:1 hr Factor:IL2v | Timepoint:1 hr Factor:RT+IL2v | Timepoint:1 hr Factor:RT+IL2v | Timepoint:1 hr Factor:RT+IL2v | Timepoint:1 hr Factor:aCD25 | Timepoint:1 hr Factor:aCD25 | Timepoint:1 hr Factor:aCD25 | Timepoint:1 hr Factor:Untrx | Timepoint:2 hr Factor:Untrx | Timepoint:2 hr Factor:Untrx | Timepoint:2 hr Factor:RT | Timepoint:2 hr Factor:RT | Timepoint:2 hr Factor:RT | Timepoint:2 hr Factor:IL2v | Timepoint:2 hr Factor:IL2v | Timepoint:2 hr Factor:IL2v | Timepoint:2 hr Factor:RT+IL2v | Timepoint:2 hr Factor:RT+IL2v | Timepoint:2 hr Factor:RT+IL2v | Timepoint:2 hr Factor:aCD25 | Timepoint:2 hr Factor:aCD25 | Timepoint:2 hr Factor:aCD25 | Timepoint:2 hr Factor:Untrx | Timepoint:6 hr Factor:Untrx | Timepoint:6 hr Factor:Untrx | Timepoint:6 hr Factor:RT | Timepoint:6 hr Factor:RT | Timepoint:6 hr Factor:RT | Timepoint:6 hr Factor:IL2v | Timepoint:6 hr Factor:IL2v | Timepoint:6 hr Factor:IL2v | Timepoint:6 hr Factor:RT+IL2v | Timepoint:6 hr Factor:RT+IL2v | Timepoint:6 hr Factor:RT+IL2v | Timepoint:6 hr Factor:aCD25 | Timepoint:6 hr Factor:aCD25 | Timepoint:6 hr Factor:aCD25 | Timepoint:6 hr Glucose_M+0 2359850 2603659 2405534 2860581 2469462 2443054 2977947 2489446 2917247 2240224 2868271 2505238 2855700 2428542 2448348 2656924 2565671 2699020 2516084 2837602 2376858 2172919 2540118 2675791 2810335 2691599 2033283 2986790 2537353 1627880 2804868 2773944 2627254 2792269 2784899 2671985 2454473 2478416 2200735 2417773 2892951 2628914 2568955 2438586 2652546 2776305 2871012 2811020 2726532 2438453 3035879 2503840 2504177 2503060 2783220 2459162 3111494 2481118 2747292 2900794 Glucose_M+6 21573520 21066010 21603360 23435140 16959410 20396900 23986350 20878080 23686560 18751080 24967430 21697180 22837630 21378030 18951770 22625470 21169810 21603810 22621870 22925420 20914090 17154280 20783450 21516880 22157380 19420940 19353930 23849950 23326430 11018340 20949300 22877720 22412610 21231810 20232500 22400340 16026250 20670730 19962370 21343690 22861380 21769300 19785870 19208430 21612570 22670450 20932560 22604870 21938630 20425810 24800390 18883300 19344700 19928550 23483690 19368760 22833710 21611280 21809690 21578930 D-Hexose-phosphate_M+0 9911 13268 19850 17687 11857 9753 5546 17178 14967 9527 20023 15817 12767 13267 11110 13125 14642 11151 20059 16902 9809 18624 15302 11742 15546 16883 11504 19636 10386 17450 8798 15071 15615 9319 15682 16125 14267 16275 14661 11511 6358 18132 13313 18972 7617 23582 11630 19021 17377 9546 15489 11768 10799 14277 17599 17694 17160 13851 15347 13607 D-Hexose-phosphate_M+6 4114 4429 5386 2701 4851 787 2670 2708 3525 2101 759 3324 3281 1732 2283 9124 8155 6311 2688 5361 7677 4946 3053 4131 4218 1936 10073 7493 6482 7385 9262 6360 10260 9489 9023 5135 4813 10262 10398 6336 5429 3742 10113 7057 6467 18341 23763 24204 16645 21034 28425 17039 28586 25424 32361 23977 21759 22528 22168 28761 D-Glyceraldehyde 3-phosphate 131312 163777 63876 184155 56822 136494 131617 91411 235351 146743 49768 73337 169959 107394 124882 194509 66514 53219 76110 69534 59425 150163 62851 124809 145001 174441 61369 57943 90421 55030 58691 100076 81073 56113 86242 87054 51694 87001 58402 162659 1239935 121025 277205 180186 126355 82196 124361 30728 92728 52634 151042 87689 79654 55759 69038 52537 63745 60419 31671 90837 Pyruvate_M+0 304847 244771 251659 277521 289562 250498 301604 225340 332061 264616 271055 278064 303060 222585 293789 303492 253704 292737 353972 335200 224575 257321 304584 286351 238549 296269 295729 252783 283416 358288 234191 271079 277421 281927 357085 259690 231486 225339 256482 272982 280572 197695 231468 291068 314002 277345 262365 360760 305363 381020 273794 295169 247932 282560 309867 300927 258217 229732 203787 303822 Pyruvate_M+3 734936 771030 694394 885837 643028 749329 814468 695813 860258 604677 855736 762002 835719 715820 711741 835278 807052 703674 819749 904501 696195 599467 773703 853959 896539 678364 692156 841117 745642 700135 709856 838476 753883 768216 853929 777432 581230 748333 619943 764674 858229 770481 690921 714624 817175 679455 750177 631542 827625 867007 812217 705668 659945 757031 817209 673738 746147 708857 804858 863003 Lactate_M+0 26620410 33550540 26943730 23550360 30893260 34474780 33929680 30122820 27998480 22622990 26450870 26645040 24552250 30333840 32805400 27708510 29709790 27932670 31876990 29870740 26367250 31197000 31185590 28415730 27821290 26824060 30436710 31794530 32866360 21920320 29352390 30617440 33155040 24096900 30009570 27876370 26825460 26676910 29627030 28443480 27318650 26626650 27666050 29492500 28596070 26480350 31298860 29582100 36327040 30676300 29287670 30684980 30690630 29239000 29897900 31719870 29924850 28161830 32317330 31958980 Lactate_M+3 12464080 12832990 12594950 13810610 9525991 11902250 13751020 11655080 13632440 10384560 14363540 12381020 12757950 12005530 11071000 13026010 12227240 12301350 13345030 13543820 11518170 9885209 12462190 12240020 13339130 10953340 11176870 13450260 13331030 8796634 11586160 12667360 12897240 12248120 13383320 12658420 9223069 11432430 11230260 11977180 12859800 12729200 11118600 10681880 13007620 11671830 12095460 13261700 12446190 11592200 13273370 11025510 10694870 11493960 14074110 10927790 12467300 11942440 12669190 12254210 Citrate_M+0 8872112 6732596 7678008 6864178 6302508 6480454 5752062 7563300 7642594 6953540 7093070 7651896 5658404 6075072 8343138 7826190 7262866 6640638 8393222 8265970 6133716 6665506 7097246 6641042 7164714 5955574 5091354 7458488 6630562 6303052 6009380 7058328 7443104 6655220 5475310 6585162 6060008 7017670 6181884 7430364 6781090 7683078 5690214 5306002 5432726 7568782 7927374 6534168 6562732 8201706 7363968 6989694 5409326 9483927 9578972 8523064 7156514 6587124 7998396 8754722 Citrate_M+2 68425 65492 79655 63790 55915 85094 54848 74711 56349 62753 73236 59088 44708 65791 77350 72037 97949 86664 91097 70952 64812 55153 60154 64983 57726 54373 47936 59774 56816 66055 64464 73596 86309 78893 68111 64551 43426 54529 60913 78524 65794 85693 39165 41160 62769 79274 85432 66169 43258 64653 69936 58536 67877 74069 88775 82805 73392 50403 74516 89276 2-Oxoglutarate_M+0 110190 122470 103168 88721 116840 151472 109579 106122 88900 86516 93598 116126 104083 108517 100120 144569 119710 130530 105886 94895 122119 125974 125750 93425 102569 123670 116756 117420 104886 142392 105190 95552 105585 108167 126440 132751 94789 98867 122528 101633 114647 104250 83169 121602 96346 97582 101443 138408 101186 165994 90349 94001 103057 145172 144547 128615 102208 127852 107952 105231 Succinate_M+0 6455244 4104897 4813419 3753077 5048950 6278284 4376498 4630286 4611708 4861722 4402163 5049958 4309426 4394049 4400522 4826440 5381030 4647664 4906664 5146340 4211556 4839708 4876776 4589748 3932630 4709902 4616820 4884934 3549952 5520458 4131535 4881800 4981770 3562602 4692334 4380998 4645456 4091843 4591509 3957300 3730316 4267378 4013379 4572296 3928431 5045594 5250816 5403772 5386988 5723020 4997461 4895154 4031453 5429792 5709142 6250176 4951040 5321198 5445221 5382260 Succinate_M+4 50652 56841 47284 45271 51651 41067 63948 44020 51037 41915 48192 43209 36328 55777 44103 64732 31860 24693 55360 34628 44999 36179 69589 57827 39832 58670 40018 43136 48856 39950 37067 45506 27030 42208 46074 36060 53002 42158 52488 58557 47685 41277 32384 48071 49380 63802 47323 72663 56644 35822 43823 62036 43449 64764 38372 67796 64765 42647 72048 63537 Itaconate_M+0 324204 262403 247596 217296 224328 263754 184122 247560 260733 235547 229071 267026 221492 253026 227434 299642 256741 265964 220336 237809 230354 269081 227179 266849 222743 244151 228917 253334 234720 463980 200776 269553 250552 248761 252313 251220 213890 256570 239584 217923 235755 252518 178552 246506 195411 242578 240763 284120 257556 279962 250441 232254 230992 286514 321587 314307 250901 245877 243587 285754 Fumarate_M+0 629001 490660 442935 410810 474726 551130 368325 430766 434655 462311 438788 511802 403842 425304 503165 540671 502392 422088 460911 467059 394791 530241 447569 439239 361834 407624 436286 480823 423282 948750 395332 459281 474937 412073 450620 474943 424346 451719 407431 409678 388341 391738 375611 472797 386426 516689 505130 536210 396316 497320 461625 457837 361005 531537 529312 541754 443743 468129 464981 539717 (S)-Malate+M+0 2700998 2265853 2083738 1555401 2024259 2646246 1802438 1885185 1995942 2081785 1780927 2348717 1844750 1941662 1665988 2732088 2522199 1995067 2270451 1962414 2004288 2570102 2094318 2043421 1433330 2070254 1928931 2212869 1713801 3308769 1870048 2139881 2176104 2060636 2261144 2309455 1905392 1917641 1967845 1967249 1984036 2103810 1708045 2288836 1873087 2248748 2251659 2651360 1621284 2245825 2256116 2049215 1543267 2537701 2143364 2393516 2061624 2212921 2313757 2178290 2-Hydroxyglutarate_M+0 417672 377611 321349 273486 336994 422975 313468 280286 314568 312106 263223 366876 330608 322930 242881 437563 381097 368888 329894 287718 309416 408430 340933 325761 233708 355488 314650 316307 332451 470141 320490 317468 317009 350252 361759 375795 283093 302182 328748 295668 341479 333101 277867 361804 285625 343553 352363 413218 340352 368595 334274 313447 309362 423601 358717 421334 333809 400108 356532 279663 2-Hydroxyglutarate_M+3 43563 81099 56494 67282 44541 37798 45971 57446 62375 52335 47546 57668 65248 47508 44061 71742 71464 60520 52224 65383 49982 49331 55416 64836 59462 49396 27114 65700 42517 66196 66908 52767 41856 63344 83583 55985 40579 51566 56833 52423 81408 52192 77051 48268 46996 34863 58284 51828 54062 32996 64238 49900 62307 61172 47645 51031 55504 55471 46961 62740 L-Aspartate_M+0 4751472 4631282 4671786 4133782 4506762 4516578 3996247 4421436 5718814 5080710 4451434 4476378 3861598 4467742 4729614 4249764 4886492 4438706 4490986 4448396 4410077 4001161 3900355 4563466 4113543 4411740 4004271 4344174 3657087 3768391 4033661 4767522 4166024 4742271 4215864 4365572 4424447 4144949 4204136 4459249 4454456 4644425 4036170 3800529 3946644 4549874 4883113 4697334 3922093 4909970 4949654 3659072 4313300 4092014 4379252 4692526 3680123 4118658 4796881 4356932 Aspartate_M+2 47184 54613 47220 29583 46417 50244 61022 55732 26114 63639 32740 60989 46449 66058 71774 48228 51345 17928 45315 48810 25108 46539 50262 69804 40996 58091 53983 49054 45453 54763 35165 52741 42967 44579 42271 44764 41914 52087 58621 45158 62057 46048 41366 60344 34818 23621 34366 52480 71248 51311 20643 52953 52396 46664 69302 68070 70470 64236 74429 43079 L-Alanine_M+0 383488 320313 330619 332683 304270 271211 347374 272745 422622 321869 366068 310479 301191 312349 342813 303336 337421 304354 357181 349206 310027 309576 296585 371484 335193 362434 288961 300150 281285 496842 278381 333258 314966 332000 313125 337967 289797 299521 268025 294672 355669 315133 315378 267771 297838 288856 337093 364495 331383 353533 359022 297657 369572 321443 287609 333855 284992 271193 367539 307086 L-Glutamate_M+0 7511484 6642690 6688526 5916950 6639866 6704984 5710974 6411914 7838102 7031626 6414820 6330940 5755466 6484766 6414448 5995026 7338078 6334052 6129084 6978806 6440948 5958578 6282226 6600534 5877750 6144356 5630050 6417186 4973742 5157725 5863678 7076662 6689662 6762802 6235266 6031486 6319694 6620536 6639876 6327692 6628350 6824560 5674888 5697086 5462536 6665278 7253460 6815868 5871756 8285960 7252444 6018622 6439372 8166444 7250602 7919308 5391086 6138366 6698174 7132430 Glutamate_M+2 119241 115951 108805 148747 95508 98120 144980 123499 112285 94781 111339 91020 125457 121035 107921 88144 96652 115626 122173 122712 97256 71228 95384 134242 132157 87479 97182 116835 145528 83499 92914 78832 137576 121937 129884 117504 91225 98164 128354 109284 135514 92874 113337 98278 134545 127252 117023 109606 104630 103666 131380 108146 100365 120515 135579 68128 100756 110807 139258 113271 Glutamate_M+3 2829351 2906167 2543296 3269895 2282987 2985165 3203842 2720250 3290596 2526330 3313137 2922444 2993917 2829985 2416894 3163454 2810890 2862536 3026312 2924202 2577187 2306492 2878851 2954309 3326974 2611814 2506546 3195279 2685953 2115835 2609171 3124108 2950567 2687754 3310230 2850668 2010260 2716380 2613295 2707283 2998488 2864561 2580335 2796457 2908899 2889641 2717627 3071391 2893362 2671262 3270079 2536085 2463550 2603587 3140365 2482191 2820419 2756097 3173937 2731382 L-Glutamine_M+0 11428200 9386733 12812030 11811700 12110610 10214550 11686830 11919660 12419840 11284050 13437130 13076150 9882493 11150400 11433260 12478660 8927763 12671500 13291610 12989850 10277620 10640980 10943450 11130980 12710520 13328370 11182380 11669450 12475260 8729225 12483010 11583100 11390130 11621860 11721630 11439220 11498710 9882472 11042810 11186750 9558138 9588021 10547960 11235820 13052270 12123400 11613800 11209720 11391310 10148350 12617770 11351260 10174180 11612550 10870330 9782903 9801097 9285739 11913500 13272040 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name KEGG ID m/z medRt Glucose_M+0 C00031 179.0547 0.6959316 Glucose_M+6 C00031 185.0751 0.6938977 D-Hexose-phosphate_M+0 C02965 259.0217 0.6368333 D-Hexose-phosphate_M+6 C02965 265.0421 0.6333548 D-Glyceraldehyde 3-phosphate C00118 168.9882 0.6980625 Pyruvate_M+0 C00022 87.007 0.6756897 Pyruvate_M+3 C00022 90.01716 0.704833 Lactate_M+0 C01432 89.02275 0.6712347 Lactate_M+3 C01432 92.03269 0.6999158 Citrate_M+0 C00158 191.0186 0.6491612 Citrate_M+2 C00158 193.0225 0.6464448 2-Oxoglutarate_M+0 C00026 145.0127 0.6157086 Succinate_M+0 C00042 117.0176 0.6388919 Succinate_M+4 C00042 121.0312 0.6809687 Itaconate_M+0 C00490 129.0177 0.6396056 Fumarate_M+0 C00122 115.002 0.6324406 (S)-Malate+M+0 C00149 133.0127 0.6264482 2-Hydroxyglutarate_M+0 C02630 147.0284 0.6251547 2-Hydroxyglutarate_M+3 C02630 150.0383 0.7028996 L-Aspartate_M+0 C00049 132.0287 0.6547608 Aspartate_M+2 C00049 134.0345 0.6780877 L-Alanine_M+0 C00041 88.03868 0.6682743 L-Glutamate_M+0 C00025 146.0443 0.649693 Glutamate_M+2 C00025 148.0503 0.6913173 Glutamate_M+3 C00025 149.0537 0.6979005 L-Glutamine_M+0 C00064 145.0605 0.6826329 METABOLITES_END #END