#METABOLOMICS WORKBENCH lgafeira_20241119_034826 DATATRACK_ID:5382 STUDY_ID:ST003584 ANALYSIS_ID:AN005887 PROJECT_ID:PR002214 VERSION 1 CREATED_ON November 21, 2024, 6:08 pm #PROJECT PR:PROJECT_TITLE Glucagon is a major controller of metabolism and malignancy in pancreatic PR:PROJECT_TITLE neuroendocrine tumour (pNET) cells PR:PROJECT_SUMMARY Cancer metabolism research has studied the relationship between cellular PR:PROJECT_SUMMARY bioenergetics, biosynthesis, and tumour progression. However, the impact of PR:PROJECT_SUMMARY systemic metabolism and diet on tumour evolution is less understood. This study PR:PROJECT_SUMMARY delves into the role of glucagon in the regulation of cancer cell features and PR:PROJECT_SUMMARY the metabolic remodelling in the presence and absence of glucose, in pancreatic PR:PROJECT_SUMMARY neuroendocrine tumours (pNETs). To fulfil the experiments pNET cell lines (BON-1 PR:PROJECT_SUMMARY and QGP-1) and non-malignant pancreatic α-TC1 cell line were used as models. PR:PROJECT_SUMMARY Results showed that pNET cells responded differently to glucose deprivation than PR:PROJECT_SUMMARY α-TC1 cells. Specifically, pNET cells upregulated GCGR in the absence of PR:PROJECT_SUMMARY glucose, while α-TC1 cells did so in high-glucose conditions. Glucagon PR:PROJECT_SUMMARY activated the MAPK/ERK pathway, especially in pNET cells without glucose and PR:PROJECT_SUMMARY α-TC1 cells with high glucose conditions. Glucagon enhanced metabolic PR:PROJECT_SUMMARY viability, proliferation, and migration in pNET cells under glucose-deprived PR:PROJECT_SUMMARY conditions and hyperglucagonaemia. Meanwhile, in α-TC1 cell line, glucagon PR:PROJECT_SUMMARY modulated these features under high-glucose conditions and physiological PR:PROJECT_SUMMARY glucagon levels. Metabolic analysis using NMR revealed differences in amino acid PR:PROJECT_SUMMARY levels and metabolic processes based on glucose availability between cell lines. PR:PROJECT_SUMMARY Furthermore, gene expression analysis showed that glucose conditions largely PR:PROJECT_SUMMARY influenced gene expression patterns in both pNETs and α-TC1. Interestingly, PR:PROJECT_SUMMARY QGP-1 and α-TC1 produced glucose in no-glucose conditions, and glucagon PR:PROJECT_SUMMARY upregulated glucose production in α-TC1. Hence gluconeogenesis may be PR:PROJECT_SUMMARY beneficial for some pNET subsets, and it may open new perspectives to novel PR:PROJECT_SUMMARY metabolism-based strategies to clinically manage pNETs. Furthermore, the PR:PROJECT_SUMMARY production of glucose by α-cells was not described so far and is a step forward PR:PROJECT_SUMMARY in endocrinology and systemic metabolism. Lastly, an association between GCGR PR:PROJECT_SUMMARY expression and malignancy was observed, despite the decreased expression in PR:PROJECT_SUMMARY tumours comparing with the normal tissue. A negative correlation between GCGR PR:PROJECT_SUMMARY and GLP-1R expression was observed, highlighting the putative biological role of PR:PROJECT_SUMMARY glucagon in pNETs malignancy. PR:INSTITUTE ITQB NOVA PR:LAST_NAME Gonçalves PR:FIRST_NAME Luís PR:ADDRESS Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal PR:EMAIL lgafeira@itqb.unl.pt PR:PHONE 214469464 PR:CONTRIBUTORS Bárbara Ferreira, Isabel Lemos, Cindy Mendes, Beatriz Chumbinho, Fernanda PR:CONTRIBUTORS Silva, Daniela Pereira, Emanuel Vigia, Luís G. Gonçalves, António Figueiredo, PR:CONTRIBUTORS Daniela Cavaco, Jacinta Serpa #STUDY ST:STUDY_TITLE Glucagon is a major controller of metabolism and malignancy in pancreatic ST:STUDY_TITLE neuroendocrine tumour (pNET) cells ST:STUDY_SUMMARY Cancer metabolism research has studied the relationship between cellular ST:STUDY_SUMMARY bioenergetics, biosynthesis, and tumour progression. However, the impact of ST:STUDY_SUMMARY systemic metabolism and diet on tumour evolution is less understood. This study ST:STUDY_SUMMARY delves into the role of glucagon in the regulation of cancer cell features and ST:STUDY_SUMMARY the metabolic remodelling in the presence and absence of glucose, in pancreatic ST:STUDY_SUMMARY neuroendocrine tumours (pNETs). To fulfil the experiments pNET cell lines (BON-1 ST:STUDY_SUMMARY and QGP-1) and non-malignant pancreatic α-TC1 cell line were used as models. ST:STUDY_SUMMARY Results showed that pNET cells responded differently to glucose deprivation than ST:STUDY_SUMMARY α-TC1 cells. Specifically, pNET cells upregulated GCGR in the absence of ST:STUDY_SUMMARY glucose, while α-TC1 cells did so in high-glucose conditions. Glucagon ST:STUDY_SUMMARY activated the MAPK/ERK pathway, especially in pNET cells without glucose and ST:STUDY_SUMMARY α-TC1 cells with high glucose conditions. Glucagon enhanced metabolic ST:STUDY_SUMMARY viability, proliferation, and migration in pNET cells under glucose-deprived ST:STUDY_SUMMARY conditions and hyperglucagonaemia. Meanwhile, in α-TC1 cell line, glucagon ST:STUDY_SUMMARY modulated these features under high-glucose conditions and physiological ST:STUDY_SUMMARY glucagon levels. Metabolic analysis using NMR revealed differences in amino acid ST:STUDY_SUMMARY levels and metabolic processes based on glucose availability between cell lines. ST:STUDY_SUMMARY Furthermore, gene expression analysis showed that glucose conditions largely ST:STUDY_SUMMARY influenced gene expression patterns in both pNETs and α-TC1. Interestingly, ST:STUDY_SUMMARY QGP-1 and α-TC1 produced glucose in no-glucose conditions, and glucagon ST:STUDY_SUMMARY upregulated glucose production in α-TC1. Hence gluconeogenesis may be ST:STUDY_SUMMARY beneficial for some pNET subsets, and it may open new perspectives to novel ST:STUDY_SUMMARY metabolism-based strategies to clinically manage pNETs. Furthermore, the ST:STUDY_SUMMARY production of glucose by α-cells was not described so far and is a step forward ST:STUDY_SUMMARY in endocrinology and systemic metabolism. Lastly, an association between GCGR ST:STUDY_SUMMARY expression and malignancy was observed, despite the decreased expression in ST:STUDY_SUMMARY tumours comparing with the normal tissue. A negative correlation between GCGR ST:STUDY_SUMMARY and GLP-1R expression was observed, highlighting the putative biological role of ST:STUDY_SUMMARY glucagon in pNETs malignancy. ST:INSTITUTE ITQB NOVA ST:LAST_NAME Gonçalves ST:FIRST_NAME Luís ST:ADDRESS Avenida Republica, Oeiras, Not USCanada, 2780-157 Oeiras, Portugal ST:EMAIL lgafeira@itqb.unl.pt ST:PHONE 214469464 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #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 - BON1_SG_C1 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_C_N1 SUBJECT_SAMPLE_FACTORS - BON1_SG_C2 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_C_N2 SUBJECT_SAMPLE_FACTORS - BON1_SG_C3 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_C_N3 SUBJECT_SAMPLE_FACTORS - BON1_SG_250_1 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_250_N1 SUBJECT_SAMPLE_FACTORS - BON1_SG_250_2 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_250_N2 SUBJECT_SAMPLE_FACTORS - BON1_SG_250_3 Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_SG_250_N3 SUBJECT_SAMPLE_FACTORS - BON1_HG_C1 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_C_N1 SUBJECT_SAMPLE_FACTORS - BON1_HG_C2 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_C_N2 SUBJECT_SAMPLE_FACTORS - BON1_HG_C3 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_C_N3 SUBJECT_SAMPLE_FACTORS - BON1_HG_250_1 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_250_N1 SUBJECT_SAMPLE_FACTORS - BON1_HG_250_2 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_250_N2 SUBJECT_SAMPLE_FACTORS - BON1_HG_250_3 Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=BON1_HG_250_N3 SUBJECT_SAMPLE_FACTORS - QGP1_SG_C1 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_C_N1 SUBJECT_SAMPLE_FACTORS - QGP1_SG_C2 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_C_N2 SUBJECT_SAMPLE_FACTORS - QGP1_SG_C3 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_C_N3 SUBJECT_SAMPLE_FACTORS - QGP1_SG_250_1 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_250_N1 SUBJECT_SAMPLE_FACTORS - QGP1_SG_250_2 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_250_N2 SUBJECT_SAMPLE_FACTORS - QGP1_SG_250_3 Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_250_N3 SUBJECT_SAMPLE_FACTORS - QGP1_HG_C1 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_C_N1 SUBJECT_SAMPLE_FACTORS - QGP1_HG_C2 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_C_N2 SUBJECT_SAMPLE_FACTORS - QGP1_HG_C3 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Species=Human; RAW_FILE_NAME(raw_files)=QGP1_SG_C_N3 SUBJECT_SAMPLE_FACTORS - QGP1_HG_250_1 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_250_N1 SUBJECT_SAMPLE_FACTORS - QGP1_HG_250_2 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_250_N2 SUBJECT_SAMPLE_FACTORS - QGP1_HG_250_3 Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Species=Human; RAW_FILE_NAME(raw_files)=QGP1_HG_250_N3 SUBJECT_SAMPLE_FACTORS - aTC1_SG_C1 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_C_N1 SUBJECT_SAMPLE_FACTORS - aTC1_SG_C2 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_C_N2 SUBJECT_SAMPLE_FACTORS - aTC1_SG_C3 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_C_N3 SUBJECT_SAMPLE_FACTORS - aTC1_SG_250_1 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_250_N1 SUBJECT_SAMPLE_FACTORS - aTC1_SG_250_2 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_250_N2 SUBJECT_SAMPLE_FACTORS - aTC1_SG_250_3 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_SG_250_N3 SUBJECT_SAMPLE_FACTORS - aTC1_HG_C1 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_C_N1 SUBJECT_SAMPLE_FACTORS - aTC1_HG_C2 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_C_N2 SUBJECT_SAMPLE_FACTORS - aTC1_HG_C3 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_C_N3 SUBJECT_SAMPLE_FACTORS - aTC1_HG_250_1 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_250_N1 SUBJECT_SAMPLE_FACTORS - aTC1_HG_250_2 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_250_N2 SUBJECT_SAMPLE_FACTORS - aTC1_HG_250_3 Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Species=mouse; RAW_FILE_NAME(raw_files)=aTC1_HG_250_N3 #COLLECTION CO:COLLECTION_SUMMARY After starvation, 1×105 cells/well/500 µL were cultured in 24-well plates for CO:COLLECTION_SUMMARY 48 h in the presence of 250 pg/mL of glucagon, without or with glucose (4.5 CO:COLLECTION_SUMMARY g/L). The supernatant was collected, cleared upon centrifugation (150 × g, 5 CO:COLLECTION_SUMMARY min) and stored at -80C until NMR analysis. CO:SAMPLE_TYPE Culture Media CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY After starvation, 1×10^5 cells/well/500 µL were cultured in 24-well plates for TR:TREATMENT_SUMMARY 48 h in the presence of 250 pg/mL of glucagon, without or with glucose (4.5 TR:TREATMENT_SUMMARY g/L). #SAMPLEPREP SP:SAMPLEPREP_SUMMARY For sample preparation, 60 µL of a solution containing 0.022 mM trimethylsilyl SP:SAMPLEPREP_SUMMARY propionate-d4 (TSP) and 0.04% (v/v) sodium azide, both in D2O, was added in a SP:SAMPLEPREP_SUMMARY 1:1 proportion to each supernatant. #ANALYSIS AN:ANALYSIS_TYPE NMR #NMR NM:INSTRUMENT_NAME Bruker UltrashiedTM 500 Plus NM:INSTRUMENT_TYPE FT-NMR NM:NMR_EXPERIMENT_TYPE 1D-1H NM:SPECTROMETER_FREQUENCY 500 NM:NMR_PROBE 5 mm prodigy TCI-z NM:NMR_SOLVENT 10 % D2O/ 90 % H20 NM:NMR_TUBE_SIZE 5 mm NM:SHIMMING_METHOD Topshim NM:CHEMICAL_SHIFT_REF_CPD TSP NM:TEMPERATURE 25 #NMR_METABOLITE_DATA NMR_METABOLITE_DATA:UNITS mM NMR_METABOLITE_DATA_START Samples BON1_SG_C1 BON1_SG_C2 BON1_SG_C3 BON1_SG_250_1 BON1_SG_250_2 BON1_SG_250_3 BON1_HG_C1 BON1_HG_C2 BON1_HG_C3 BON1_HG_250_1 BON1_HG_250_2 BON1_HG_250_3 QGP1_SG_C1 QGP1_SG_C2 QGP1_SG_C3 QGP1_SG_250_1 QGP1_SG_250_2 QGP1_SG_250_3 QGP1_HG_C1 QGP1_HG_C2 QGP1_HG_C3 QGP1_HG_250_1 QGP1_HG_250_2 QGP1_HG_250_3 aTC1_SG_C1 aTC1_SG_C2 aTC1_SG_C3 aTC1_SG_250_1 aTC1_SG_250_2 aTC1_SG_250_3 aTC1_HG_C1 aTC1_HG_C2 aTC1_HG_C3 aTC1_HG_250_1 aTC1_HG_250_2 aTC1_HG_250_3 Factors Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:BON1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:BON1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:No | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:No Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:QGP1 pancreatic cancer cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:No | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:No Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Sample source:aTC1 pancreatic non-malignant immortalized cells | Added Glucose:Yes | Added Glucagon:Yes Alanine 93.5 93.7 93.6 93.7 87.1 92.8 315.8 234.3 265.9 243.6 250.3 247.25 0 0 0 41.3 58.8 49.25 93.6 199.8 146.4 121.2 115 117.5 0 0 0 0 0 0 70 76.3 72.3 83.5 85 83.6 Arginine 0 0 0 0 0 0 845.4 513 681.3 485.4 471 478.5 0 0 0 0 0 0 357.7 327.9 342.1 526 615.9 569.2 0 0 0 0 0 0 433.4 217 326.9 357.7 165.6 261.2 Glutamine 0 0 0 0 0 0 163.3 122.6 139.1 0 0 0 0 0 0 0 0 0 448.4 373.9 411.9 1081 931.3 1005.2 0 0 0 0 0 0 0 0 0 0 0 0 Glycine 299.2 299.5 299.35 279.2 265.7 298.5 278 197.5 245.2 186.3 172.2 178.9 131 130.5 130.3 39.4 30.3 35.9 104.7 89.1 97.1 135.9 121.3 128.1 234.4 81.3 157.15 117.8 489.1 303.2 152.7 190.3 170.2 173.7 156.7 165.8 Isoleucine 267.1 292.8 279.95 327.7 309.3 280.8 345.1 280.9 307.8 304.8 263.5 284.6 517.3 489.6 503.1 450.4 521.1 484.6 269.3 236 252.1 287.5 303.4 294.9 343.5 255 300.1 433.4 586.6 509.7 460.1 538.5 498.1 399.7 330.9 366.7 Leucine 263.2 266 264.6 325.4 259.8 265.3 329 295.6 309.5 309.6 291.5 300.8 548.2 402.8 476.1 384 170 275.6 278.4 195.5 237.3 251.7 281.7 265.5 681.7 319.4 500.85 373.6 373.8 373.3 474.8 587 529.1 608.5 548.4 577.2 Lysine 374.6 379.6 377.1 419.2 394.2 378.4 728.8 449.8 572.6 289.4 289.7 289.9 471 437.2 454.9 370 372.9 369.1 100.6 145.9 122.6 92.2 95.7 94.2 690.3 373 531.1 630.9 682.6 655.5 256 186 220.6 224.6 432.7 329.9 Methionine 91.5 86.7 89.1 106 106.1 88.5 165.1 101.3 131.5 76.7 78.2 77.9 131.8 156.9 144.7 108.6 139.8 125.5 50.2 55.3 53.1 28.6 36.5 32.1 207.8 165.8 187.3 181.3 184.7 183.5 174.5 95.9 136.7 146.3 140.5 142.8 Phenylalanine 201.6 203.8 202.7 121 119.4 203.2 233 180.8 207.6 138.9 184 161.8 320.1 274.6 297.9 346.4 373.6 363.2 63.3 66.2 65.2 89.5 69.9 80.3 164.7 274.6 218.95 226.6 230.9 228.15 300.4 377.9 338.4 140.1 240.5 189.1 Proline 176.1 168.6 172.35 144.6 141.3 173.9 205.6 156.7 179.2 119.6 389.3 254.2 33.4 37.6 35.8 24.9 13.6 18.5 267.4 163.5 215.1 0 0 0 28.6 24.3 27.15 24.3 23 23.2 0 0 0 0 0 0 Pyroglutamate 52.6 52 52.3 116.6 113.5 51.9 2327 1374.7 1844.1 1453.6 1446.5 1450.2 30.6 90.1 60.9 45.1 75.6 61.6 855.7 1006.2 930.4 906.7 899.7 902.4 18.3 38.5 29.2 88.8 88.3 88.1 815.7 811.8 814.2 1090.8 1393.2 1243.5 Serine 42.2 42.2 42.2 34.6 31.7 43.8 185.3 211.1 204.3 139.4 192 165.3 78.8 63.9 71.7 31.4 31.4 32.7 212.7 101.5 157.7 136.3 134.4 135.9 425.4 84.7 254.75 37.6 41.3 39.8 0 0 0 92.1 107.5 99.3 Threonine 415.1 402.6 408.85 549.8 335 407.1 587.5 317.3 467.5 293.1 315.7 304.9 443.3 335.3 389.8 493.1 500.3 495.2 87 101.6 94.9 141.3 94.3 117.2 736.7 523.3 628.8 609.1 673.9 642.1 657.9 586.4 621.3 565.7 507.7 535.5 Tryptophan 42.5 30.6 36.55 38.9 33.1 35.9 33.6 33.5 34 27.6 28 27.5 48.3 30.6 39.8 51.9 51.6 53.4 11.5 12.3 11.4 17.8 18.4 18.8 49.3 37 44.05 48.7 50.4 49.8 62.5 58.1 61.8 54.8 42.5 47.8 Tyrosine 141.5 149.8 145.65 0 0 0 202 152.8 200 0 0 0 0 0 0 0 0 0 76 77.2 76.1 83.7 83.2 84.1 0 0 0 297.6 304 300.2 307.1 314 309.2 267.3 300 283.1 Valine 254.7 253 253.85 321.4 238.2 252.4 449.4 318 378 206.2 281.1 243.4 448.3 399.5 424.6 489.8 494.8 494.9 85.6 91.6 84.2 131.5 125.1 128.7 506.3 256.1 382.4 502.1 505.5 503.5 596.2 588.4 592.3 637.9 540.8 589.35 Acetate 817.5 713.8 765.65 782.6 782.4 766.1 946.6 667.8 816.8 621.2 589.3 605.55 66 66.2 65.6 76.8 75.3 77.9 61.9 65.3 63.2 50.9 80.7 65.1 475.4 475.8 476.2 474.6 481.1 477.5 378.4 237.1 308.5 242.3 265 252.2 Formate 45.4 40.7 43.05 34.3 34.7 43.7 32 20.6 30.1 22.9 20.4 21.3 23.3 22.1 23.1 23.2 24.3 23.3 21.3 39.5 30.7 29.6 25.8 27.1 12.2 18.7 15.2 16.9 16.8 16.6 24.5 15.9 19.5 14.6 17.9 15.8 Isobutyrate 140 143.7 141.85 186.5 183.1 142.2 7.6 4.3 6.2 0 0 0 268.3 236.2 252.8 261.5 261.7 260.4 58 55.1 57.2 59 60.9 60.8 0* 258.9 130.5 253.1 274 264.05 374.5 328.9 352.9 343.9 341 342.9 Lactate 658.2 777.1 717.65 1568.9 735.5 716.9 3567 2312 2943.2 2433.2 2335.7 2384.9 307.6 345.1 327.9 446.7 363.8 404.1 672.3 770 720.3 964.3 948 957.8 443 402.4 421.8 438.5 441.9 440.8 295.6 231.8 262.4 333.4 309.4 320.5 Nicotinurate 0 0 0 0 0 0 18 8.8 15.8 14.5 14.4 14.6 0 0 0 0 0 0 6.9 7.5 7.8 5.9 6.5 6.8 19.3 7.7 14.1 26.9 25.1 25.6 25.3 25.6 26.8 19.7 24.3 21.4 Pyruvate 64.6 62.1 63.35 95 57.9 62.9 235 183.6 211.8 161.4 165.2 163.9 469 455.3 462.8 596.3 513.4 556.3 78.5 117.6 97.8 95.4 87.9 91.2 422.9 395.8 409.75 366.6 477.6 421.6 230.2 190.5 211.9 201.1 201.8 200.2 Succinate 11 6.1 8.55 19.8 13.3 9.2 0 0 0 0 0 0 11.1 13.5 12.7 3.4 2.3 3.1 0 0 0 0 0 0 12.8 20.6 16.2 8.8 19.3 14.3 286.4 237.5 260.1 0 0 0 Choline 51.1 51.3 51.2 56.9 50.4 51.8 87.5 45.7 63.8 47 42.1 44.8 22.3 22.7 22.2 24.5 23.8 25.3 0* 12.9 12.5 25.8 20.8 24.6 32.2 22.1 26.85 25.6 27.2 26.8 28.7 24.8 27.1 39.4 54.1 47.3 Hypoxanthine 0 0 0 0 0 0 8.5 5.9 8.7 5.7 6.3 6.2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3-Methylxanthine 5.6 4.4 5 9.9 9 5.3 3.7 2.6 4.25 0 0 0 7.8 7.5 7.9 4.7 5.8 4.85 1.5 1.5 1.2 0 0 0 8.3 8.4 8.55 9.1 8.5 8.5 3.9 6.8 6.7 2.1 2.6 2.9 Niacinamide 18.2 18.2 18.2 26.2 12.2 17.9 15.8 9.9 14.6 9.3 14.8 12.2 17.3 23.2 20.9 19.4 19.3 20.9 7 5.8 6.8 0 0 0 20.5 20.5 20.1 20.2 22 21.8 24.4 24.3 23.5 23.8 23.2 23.2 Tyramine 0 0 0 194.3 189.7 146.8 232 163.3 201.3 161.2 143.8 152.9 236.9 200.7 218.5 281.3 298.8 292.8 0 0 0 0 0 0 352.4 285.4 317.6 0 0 0 352.2 329 341.5 300 356.9 327.5 Glucose 0 0 0 0 0 0 13133.23 13133.23 13133.23 10475.57 10475.57 10475.57 196.5 260.6 228.9 260.1 260.6 262.8 5230.1 5962 5596.7 6241 5920.7 6078.3 474.2 679 577.2 981.1 964.4 972.95 19633.1 17258 18446.8 16171.7 15975.6 16073.2 NMR_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name HMDB PubChem ChEBI KEGG METLIN Alanine HMDB0000161 5950 16977 C00041 NA Arginine HMDB0000517 6322 16467 C00062 5502 Glutamine HMDB0000641 5961 18050 C00064 5614 Glycine HMDB0000123 750 15428 C00037 20 Isoleucine HMDB0000172 6306 17191 C00407 5193 Leucine HMDB0000687 6106 15603 C00123 24 Lysine HMDB0000182 5962 18019 C00047 5200 Methionine HMDB0000696 6137 16643 C00073 5664 Phenylalanine HMDB0000159 6140 17295 C00079 28 Proline HMDB0000162 145742 17203 C00148 29 Pyroglutamate HMDB0000267 7405 18183 C01879 3251 Serine HMDB0000187 5951 17115 C00065 5203 Threonine HMDB0000167 6288 16857 C00188 32 Tryptophan HMDB0000929 6305 16828 C00078 5879 Tyrosine HMDB0000158 6057 17895 C00082 34 Valine HMDB0000883 6287 16414 C00183 5842 Acetate HMDB0000042 176 15366 C00033 3206 Formate HMDB0304356 283 15740 NA NA Isobutyrate HMDB0001873 6590 16135 C02632 106 Lactate HMDB0000190 107689 422 C00186 5205 Nicotinurate HMDB0003269 68499 7563 C05380 1499 Pyruvate HMDB0000243 1060 32816 C00022 117 Succinate HMDB0000254 1110 15741 C00042 114 Choline HMDB0000097 305 15354 C00114 56 Hypoxanthine HMDB0000157 790 17368 C00262 83 3-Methylxanthine HMDB0001886 70639 62207 C16357 2820 Niacinamide HMDB0001406 936 17154 C00153 1497 Tyramine HMDB0000306 5610 15760 C00483 60 Glucose HMDB0000122 5793 15903 C00031 133 METABOLITES_END #END