#METABOLOMICS WORKBENCH hormel101_20180813_073038_mwtab.txt DATATRACK_ID:1473 STUDY_ID:ST001036 ANALYSIS_ID:AN001697 PROJECT_ID:PR000693 VERSION 1 CREATED_ON August 15, 2018, 12:40 pm #PROJECT PR:PROJECT_TITLE Mayo Pilot and Feasibility: The pathogenic NMO IgG dysregulates the astrocytic PR:PROJECT_TITLE glutamine-glutamate cycle: a metabolic basis for depression in NMO patients PR:PROJECT_SUMMARY Neuromyelitis optica (NMO) is a disabling central nervous system (CNS) PR:PROJECT_SUMMARY inflammatory disorder that involves a pathogenic autoantibody (NMO IgG) directed PR:PROJECT_SUMMARY against aquaporin-4, the major brain water channel, expressed on astrocytes. PR:PROJECT_SUMMARY Astrocytes in brain tissue from patients with NMO exhibit a spectrum of PR:PROJECT_SUMMARY abnormalities and pathologies ranging from sublytic gliosis and reactivity to PR:PROJECT_SUMMARY outright destruction. Our current working model for NMO pathogenesis involves an PR:PROJECT_SUMMARY early and robust NMO IgG-induced astrocytic stress response that drives PR:PROJECT_SUMMARY metabolic dyshomeostasis and the production of pro-inflammatory cytokines and PR:PROJECT_SUMMARY chemokines that amplify pathology by recruiting innate immune effector cells to PR:PROJECT_SUMMARY the CNS. Notably, astrocytes are implicated in clinical depression and patients PR:PROJECT_SUMMARY with NMO experience depression at levels that exceed the general population. PR:PROJECT_SUMMARY Preliminary magnetic resonance spectroscopy data from our group indicates that PR:PROJECT_SUMMARY glutamate levels are reduced in the prefrontal cortex of NMO patients, PR:PROJECT_SUMMARY suggesting that unipolar depression in these individuals is a direct pathogenic PR:PROJECT_SUMMARY effect of NMO IgG-induced astrocytic dysregulation. Because astrocytes are PR:PROJECT_SUMMARY critical for glutamine-glutamate cycling in the CNS, we hypothesize that PR:PROJECT_SUMMARY stimulation of primary astrocytes with patient-derived NMO IgG will drive a PR:PROJECT_SUMMARY metabolic shift marked by alterations in cellular levels of glutamate and PR:PROJECT_SUMMARY glutamine. In preliminary experiments using 1H-NMR to measure metabolic changes PR:PROJECT_SUMMARY induced in astrocytes by stimulation with NMO IgG we observed variable glutamate PR:PROJECT_SUMMARY and glutamine responses. To overcome issues of signal-to-noise and the high PR:PROJECT_SUMMARY basal levels of glutamate and glutamine produced by astrocytes, we now propose PR:PROJECT_SUMMARY to use isotopic tracing and 13C-NMR to quantify NMO IgG-induced metabolic PR:PROJECT_SUMMARY dysregulation. Our ultimate goal is to use NMO IgG-induced dyshomeostasis as a PR:PROJECT_SUMMARY microscope to reveal basic mechanisms of pathogenic glutamate-glutamine PR:PROJECT_SUMMARY metabolism in astrocytes that may not only impact the health of patients with PR:PROJECT_SUMMARY NMO but may also yield novel insights into the mechanisms of depression in PR:PROJECT_SUMMARY general. PR:INSTITUTE Mayo Clinic PR:LAST_NAME Howe PR:FIRST_NAME Charles PR:ADDRESS 200 First St. SW, Rochester, Minnesota, 55905, USA PR:EMAIL howe@mayo.edu PR:PHONE 507-284-9288 #STUDY ST:STUDY_TITLE TCA Concentrations in Neuromyelitis Optica Patients (part - II) ST:STUDY_SUMMARY Patients with an inflammatory disease of the central nervous system known as ST:STUDY_SUMMARY neuromyelitis optica (NMO) experience increased levels of depression. These ST:STUDY_SUMMARY patients have an antibody that recognizes a type of cell in the brain called ST:STUDY_SUMMARY astrocytes – binding of this antibody to astrocytes triggers a stress response ST:STUDY_SUMMARY in the cell that results in the development of brain lesions that cause ST:STUDY_SUMMARY disability and cognitive disturbances. We recently observed a change in the ST:STUDY_SUMMARY level of glutamate in a part of the brain involved in depression in patients ST:STUDY_SUMMARY with NMO. Glutamate is a chemical that is used in the brain for communication ST:STUDY_SUMMARY between neurons – reduced levels of glutamate are thought to trigger ST:STUDY_SUMMARY depression by reducing neuronal activity in specific circuits. Based on this ST:STUDY_SUMMARY observation and the known role of astrocytes in maintaining glutamate levels in ST:STUDY_SUMMARY the brain, we hypothesize that the NMO antibody disturbs metabolic activity in ST:STUDY_SUMMARY astrocytes and thereby reduces glutamate and triggers depression. We intend to ST:STUDY_SUMMARY measure TCA concentration in NMO astrocytes. It is our hope that we will not ST:STUDY_SUMMARY only learn something about the mechanisms of astrocyte dysregulation in ST:STUDY_SUMMARY neuromyelitis optica, but that we will learn something about the mechanisms of ST:STUDY_SUMMARY depression in general that may lead to new therapies for this disease. ST:INSTITUTE Mayo Clinic ST:LAST_NAME Howe ST:FIRST_NAME Charles ST:ADDRESS 200 First St. SW, Rochester, Minnesota, 55905, USA ST:EMAIL howe@mayo.edu ST:PHONE 507-284-9288 #SUBJECT SU:SUBJECT_TYPE Human SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - ms6923-1 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn protein ug/vial=505 SUBJECT_SAMPLE_FACTORS - ms6923-2 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn protein ug/vial=461 SUBJECT_SAMPLE_FACTORS - ms6923-3 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn protein ug/vial=596 SUBJECT_SAMPLE_FACTORS - ms6923-4 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn protein ug/vial=427 SUBJECT_SAMPLE_FACTORS - ms6923-5 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn protein ug/vial=459 SUBJECT_SAMPLE_FACTORS - ms6923-6 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn protein ug/vial=475 SUBJECT_SAMPLE_FACTORS - ms6923-7 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn protein ug/vial=503 SUBJECT_SAMPLE_FACTORS - ms6923-8 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn protein ug/vial=458 SUBJECT_SAMPLE_FACTORS - ms6923-9 Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn protein ug/vial=549 SUBJECT_SAMPLE_FACTORS - ms6923-10 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-11 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-12 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-13 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-14 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-15 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-16 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-17 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-18 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-19 Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 0.5 mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-20 Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-21 Time Point:0h | Matrix:Fresh Media | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-22 Time Point:0h | Matrix:Fresh Media | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-23 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-24 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-25 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6923-26 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-36 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn protein ug/vial=503 SUBJECT_SAMPLE_FACTORS - ms6982-37 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn protein ug/vial=508 SUBJECT_SAMPLE_FACTORS - ms6982-38 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn protein ug/vial=517 SUBJECT_SAMPLE_FACTORS - ms6982-39 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn protein ug/vial=492 SUBJECT_SAMPLE_FACTORS - ms6982-40 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn protein ug/vial=553 SUBJECT_SAMPLE_FACTORS - ms6982-41 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn protein ug/vial=468 SUBJECT_SAMPLE_FACTORS - ms6982-42 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGP50uMGt Gn protein ug/vial=466 SUBJECT_SAMPLE_FACTORS - ms6982-43 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGP50uMGt Gn protein ug/vial=533 SUBJECT_SAMPLE_FACTORS - ms6982-44 Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGP50uMGt Gn protein ug/vial=539 SUBJECT_SAMPLE_FACTORS - ms6982-45 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-46 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-47 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-48 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-49 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-50 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-51 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-52 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-53 Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-54 Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 0.5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-55 Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 5mM Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-56 Time Point:0h | Matrix:Fresh Media | Condition:BGP 50uMGt Gn protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-57 Time Point:0h | Matrix:Fresh Media | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-58 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-59 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- SUBJECT_SAMPLE_FACTORS - ms6982-60 Time Point:23h | Matrix:Spent Sup | Condition:E Med protein ug/vial=- #COLLECTION CO:COLLECTION_SUMMARY 4,000,000 CLOD cells collected by methanol scraping. 1ml sup also collected. CO:COLLECTION_SUMMARY There was no tracer element added to the sample. Also methanol extarction was CO:COLLECTION_SUMMARY done in ice. After the extraction samples were kept in dry ice and delivered to CO:COLLECTION_SUMMARY core. CO:SAMPLE_TYPE astrocytes #TREATMENT TR:TREATMENT_SUMMARY Murine derived pure cortical astrocytes were plated at 4x10^6 density in 10cm TR:TREATMENT_SUMMARY petri dish and grown in astro growth medium (AGM) for 2-3 days until it reaches TR:TREATMENT_SUMMARY the 90% confluency. Day before the treatment with glutamine or glutamate AGM was TR:TREATMENT_SUMMARY changed to serum free “E” media. In case of isotomer analysis 40% of glucose TR:TREATMENT_SUMMARY was switched with U13C6 (glucose) and added to serum free “E” media. On the TR:TREATMENT_SUMMARY day of treatment first the spent media was collected and then washed with base TR:TREATMENT_SUMMARY media. After washing, astrocytes were exposed to different ratio of glutamate: TR:TREATMENT_SUMMARY glutamine for 1h under non-CO2 condition. 1h spent media was collected. After TR:TREATMENT_SUMMARY washing three times with phosphate buffer saline cell pellet was collected in TR:TREATMENT_SUMMARY ice cold 100% methanol by scrapping. All the samples were immediately TR:TREATMENT_SUMMARY transferred to -70C for future analysis. E media composition: DMEM (w/o phenol TR:TREATMENT_SUMMARY red)+ 1% PenStrep+ 10% FBS+ Glutamate+ Glucose Abbreviation: Gn-Glutamine TR:TREATMENT_SUMMARY Gt-Glutamate B- Base media G- Glucose P- Sodium pyruvate #SAMPLEPREP SP:SAMPLEPREP_SUMMARY TCA concentrations of Astrocytes We intend to test the experimental hypothesis SP:SAMPLEPREP_SUMMARY that stimulation of primary murine astrocytes with patient-derived NMO IgG will SP:SAMPLEPREP_SUMMARY drive a metabolic shift marked by alterations in cellular levels of glutamate SP:SAMPLEPREP_SUMMARY and glutamine. To do so we will use isotopic tracing to measure glutamate and SP:SAMPLEPREP_SUMMARY glutamine levels in cell extracts following stimulation with NMO IgG. The cell SP:SAMPLEPREP_SUMMARY preparations, patient-derived antibody preps, isotopic tracer incubations, and SP:SAMPLEPREP_SUMMARY cell stimulations will be performed in the PI’s lab. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE GC CH:INSTRUMENT_NAME Agilent 7890B CH:COLUMN_NAME Agilent HP5-MS (30m × 0.25mm, 0.25 um) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:MS_COMMENTS - MS:INSTRUMENT_NAME Agilent 5977A MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nmol/vial MS_METABOLITE_DATA_START Samples ms6923-1 ms6923-2 ms6923-3 ms6923-4 ms6923-5 ms6923-6 ms6923-7 ms6923-8 ms6923-9 ms6923-10 ms6923-11 ms6923-12 ms6923-13 ms6923-14 ms6923-15 ms6923-16 ms6923-17 ms6923-18 ms6923-19 ms6923-20 ms6923-21 ms6923-22 ms6923-23 ms6923-24 ms6923-25 ms6923-26 ms6982-36 ms6982-37 ms6982-38 ms6982-39 ms6982-40 ms6982-41 ms6982-42 ms6982-43 ms6982-45 ms6982-46 ms6982-47 ms6982-48 ms6982-49 ms6982-50 ms6982-51 ms6982-52 ms6982-53 ms6982-54 ms6982-55 ms6982-56 ms6982-57 ms6982-58 ms6982-59 Factors Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 0.5 mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGPGt 5mM Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn Time Point:23h + 1h nonCO2 | Matrix:Pellet | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5 mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 0.5 mM Gn Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 5mM Gn Time Point:0h | Matrix:Fresh Media | Condition:BGP50uMGt Gn Time Point:0h | Matrix:Fresh Media | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 0.5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGPGt 5mM Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGP50uMGt Gn Time Point:23h +1h nonCO2 | Matrix:pellet | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 0.5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGPGt 5mM Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:1h nonCO2 | Matrix:Spent Sup | Condition:BGP50uMGt Gn Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 0.5mM Gn Time Point:0h | Matrix:Fresh Media | Condition:BGPGt 5mM Gn Time Point:0h | Matrix:Fresh Media | Condition:BGP 50uMGt Gn Time Point:0h | Matrix:Fresh Media | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Time Point:23h | Matrix:Spent Sup | Condition:E Med Lactate 55.96 37.19 57.68 52.08 51.73 57.26 26.35 29.48 53.12 102 124 140 105 131 107 107 94 78 6 6 4 6 8324 8112 9714 8991 6.61 6.83 5.7 4.26 5.04 9.2 7.44 6.27 223 273 312 219 226 341 309 326 533 44 47 44 45 9082 8234 Succinate 1.69 0.77 1.23 1.09 1.27 1.66 0.91 0.89 2.63 2 1.4 1.8 1.8 4.2 1.8 1.6 3.8 1.6 1.8 1.6 1.8 1.8 2.2 2 3.2 3.4 0.1 0.12 0.1 0.14 0.14 0.12 0.17 0.12 1.26 1.42 0.99 1.54 1.14 1.48 1.14 1.33 1.35 0.83 1.15 0.84 2.22 1.45 1.58 Fumarate 5.01 2.1 2.49 2.55 2.47 2.49 2 2.12 2.45 2.4 1 1.8 1.6 5.6 1.2 1 1.4 0.8 0.8 0.8 1 0.8 1 1 2 6 0.27 0.25 0.19 0.27 0.28 0.27 0.26 0.25 0.52 0.68 0.35 0.61 0.52 0.9 0.35 0.47 0.42 0.29 0.3 0.2 0.78 0.82 0.53 Oxaloacetate NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Ketoglutarate 5.83 3.38 5.95 6.28 4.84 5.66 4.47 5.3 6.01 0.4 0.4 0.4 0.4 0.6 0.4 0.4 0.4 0.4 0.2 0.2 0.2 0.2 2.2 2.2 2.8 2.6 0.9 1.41 1 1.12 1.1 0.89 1.42 1.27 0.52 0.41 0.56 0.47 0.58 0.68 0.53 0.66 0.65 7.05 0.12 3.08 0.21 1.6 1.48 Malate 10.11 5.6 5.81 6.05 5.57 6.28 5.36 5.74 6.38 1 0.4 0.6 0.6 1.2 0.6 0.6 1 0.4 0.4 0.2 0.4 0.4 1.4 1.4 1.8 1.8 1.4 1.45 1.17 1.61 1.64 1.2 1.54 1.43 0.76 0.36 0.47 0.4 0.46 0.62 0.44 0.52 0.56 0.23 0.22 0.25 0.54 1.38 1.11 Aspartate 32.8 16.84 23.14 40.2 35.25 37.13 34.3 33.66 35.44 1.4 1.8 1.6 1.6 1.4 1 2 2 2 2.6 1.4 1.8 2 1.4 0.8 1 1 3.66 3.76 8.34 10.35 8.41 5.73 5.64 17.12 0.61 2.06 0.82 1.58 1.23 4.49 2.4 3.17 2.59 1.21 2.07 2.39 1.16 1.08 2-Hydroxyglutarate 1.3 0.97 1.09 1.06 0.98 1.03 0.92 0.89 1.13 9.4 9.4 9.4 9.6 9.4 9.2 9.4 9.4 9.6 10.2 10 9.8 10.6 11.2 10.8 11 9.6 0.04 0.07 0.02 0.03 0.05 0.01 0.04 0.05 Glutamate 113.1 66.4 78.9 114 109.2 112.8 88.8 101 105.4 0.63 0.48 0.45 1.03 1.14 0.93 0.75 0.96 0.78 4.19 7.33 38.21 3.94 8.64 1.07 4 0.39 17.21 20.6 28.82 32.97 29.67 35.02 30.99 1.59 0.37 0.32 1.03 0.95 1.04 3.63 1.31 0.43 5.16 5.51 52.92 7.47 0.48 0.36 Glutamine 158.9 131.94 150.05 172.07 184.18 170.16 138.6 138.08 160.17 392.8 414.7 409.91 2324.59 2284.22 1920.43 445.41 423.16 378.27 375.62 2208.77 407.51 134.01 164.86 202.12 175.1 1.93 346 596 4270 4430 5230 428 428 465 405 4205 413 408 362 cis-Aconitic Acid 0.218 0.154 0.182 0.162 0.114 0.129 0.109 0.102 0.143 0.33 0.44 0.38 0.17 0.65 0.13 0.23 0.43 0.24 0.27 0.25 0.21 0.08 1.3 1.53 1.64 0.84 0.64 0.33 0.63 0.23 0.27 0.25 0.27 2.56 0.5 1.46 0.71 1.93 0.75 0.75 0.51 0.65 0.56 0.54 0.66 0.21 3.64 2.49 Citrate 25.17 14.45 19.87 17.25 15.99 17.18 16.78 16.54 20.62 8.2 7.8 9.2 7 4.4 6.6 7.6 7.2 6.6 1.2 1.2 0.8 0.4 224.6 246.8 281.2 257.8 2.7 3.09 3.32 2.89 2.73 3.07 3.18 4.56 5.93 8.19 5.92 6.71 9.12 7.24 10.29 7.56 280.59 251.87 Isocitrate 0.53 0.3 0.42 0.39 0.35 0.38 0.36 0.35 0.42 0.4 0.4 0.4 0.2 0.6 0.2 0.2 0.4 0.2 0.4 0.4 0.2 1 1.2 1.2 1 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Lactate Succinate Fumarate Oxaloacetate Ketoglutarate Malate Aspartate 2-Hydroxyglutarate Glutamate Glutamine cis-Aconitic Acid Citrate Isocitrate METABOLITES_END #END