{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003262","ANALYSIS_ID":"AN005346","PROJECT_ID":"PR002024","VERSION":"1","CREATED_ON":"June 17, 2024, 9:54 am"},

"PROJECT":{"PROJECT_TITLE":"A Covalent Creatine Kinase Inhibitor Ablates Glioblastoma Migration and Sensitizes Tumors to Oxidative Stress","PROJECT_TYPE":"LC-MS/MS","PROJECT_SUMMARY":"Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, diffuse infiltration, and near-uniform lethality. The underlying mechanisms are unknown, and no treatment has been curative. Using a recently developed kinase inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. While CKi minimally impacted GBM cell proliferation and viability, it significantly affected migration. In established GBM cell lines and patient-derived xenografts, CKi ablated both the migration and invasion of GBM cells. CKi also hindered radiation-induced migration. RNA-seq revealed a decrease in invasion-related genes, with an unexpected increase in glutathione metabolism and ferroptosis protection genes post-CKi treatment. The effects of CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 metabolite tracing indicated heightened glutathione biosynthesis post-CKi treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis activation abrogated cell survival. Our data demonstrated that CKi perturbs promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase axis as a druggable target for GBM treatment.","INSTITUTE":"Northwestern University, Feinberg School of Medicine","DEPARTMENT":"Neurological Surgery","LABORATORY":"Jason Miska","LAST_NAME":"Miska","FIRST_NAME":"Jason","ADDRESS":"676 N St. Clair","EMAIL":"jason.miska@northwestern.edu","PHONE":"8478678201"},

"STUDY":{"STUDY_TITLE":"A Covalent Creatine Kinase Inhibitor Ablates Glioblastoma Migration and Sensitizes Tumors to Oxidative Stress.","STUDY_TYPE":"Cki treatment on glioblastoma","STUDY_SUMMARY":"Glioblastoma is a Grade 4 primary brain tumor defined by therapy resistance, diffuse infiltration, and near-uniform lethality. The underlying mechanisms are unknown, and no treatment has been curative. Using a recently developed kinase inhibitor (CKi), we explored the role of this inhibitor on GBM biology in vitro. While CKi minimally impacted GBM cell proliferation and viability, it significantly affected migration. In established GBM cell lines and patient-derived xenografts, CKi ablated both the migration and invasion of GBM cells. CKi also hindered radiation-induced migration. RNA-seq revealed a decrease in invasion-related genes, with an unexpected increase in glutathione metabolism and ferroptosis protection genes post-CKi treatment. The effects of CKi could be reversed by the addition of cell-permeable glutathione. Carbon-13 metabolite tracing indicated heightened glutathione biosynthesis post-CKi treatment. Combinatorial CKi blockade and glutathione inhibition or ferroptosis activation abrogated cell survival. Our data demonstrated that CKi perturbs promigratory and anti-ferroptotic roles in GBM, identifying the creatine kinase axis as a druggable target for GBM treatment.","INSTITUTE":"Northwestern University, Feinberg School of Medicine","DEPARTMENT":"Neurological Surgery","LABORATORY":"Jason Miska","LAST_NAME":"Miska","FIRST_NAME":"Jason","ADDRESS":"676 N St. Clair","EMAIL":"jason.miska@northwestern.edu","PHONE":"8478678201","NUM_GROUPS":"2","TOTAL_SUBJECTS":"12"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-01",
"Factors":{"Sample source":"Brain","Treatment":"CTL"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-01"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-02",
"Factors":{"Sample source":"Brain","Treatment":"CTL"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-02"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-03",
"Factors":{"Sample source":"Brain","Treatment":"CTL"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-03"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-04",
"Factors":{"Sample source":"Brain","Treatment":"1hr"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-04"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-05",
"Factors":{"Sample source":"Brain","Treatment":"1hr"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-05"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-06",
"Factors":{"Sample source":"Brain","Treatment":"1hr"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-06"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-07",
"Factors":{"Sample source":"Brain","Treatment":"4hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-07"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-08",
"Factors":{"Sample source":"Brain","Treatment":"4hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-08"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-09",
"Factors":{"Sample source":"Brain","Treatment":"4hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-09"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-10",
"Factors":{"Sample source":"Brain","Treatment":"8hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-10"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-11",
"Factors":{"Sample source":"Brain","Treatment":"8hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-11"}
},
{
"Subject ID":"-",
"Sample ID":"Les-Jas-20230207-12",
"Factors":{"Sample source":"Brain","Treatment":"8hrs"},
"Additional sample data":{"RAW_FILE_NAME":"Les-Jas-20230207-12"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"GBM cells were cultured under the conditions indicated in the manuscript, then metabolites were collected using 80% methanol extraction.","SAMPLE_TYPE":"Brain"},

"TREATMENT":{"TREATMENT_SUMMARY":"metabolites isolated from Cki treated cells. Samples 1-3 are control treatment, Sample 4-6 are 1 hour CKi treatment, Samples 7-9 are 4 hours CKi treatmet, and 10-12 are 8 hours CKi treatment."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Isolated samples were dried using a SpeedVac. Acetonitrile (50%) was added to the tube for reconstitution following overtaxing for 30s. Sample solution was then centrifuged for 15min at 20,000g and 4°C. Supernatant was collected for LC-MS analysis."},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Samples were analyzed by high-performance LC (HPLC) and high-resolution MS and MS/MS (HPLC-MS/MS). The system consists of Thermo Q Exactive with an electrospray source and an UltiMate3000 (Thermo Fisher Scientific) series HPLC consisting of a binary pump, degasser, and autosampler outfitted with an XBridge Amide column (Waters; dimensions of 4.6 mm by 100 mm and a 3.5-μm particle size). The mobile phase A contained 95% water/5% acetonitrile (v/v), 20 mM ammonium hydroxide, and 20 mM ammonium acetate (pH 9.0); phase B was 100% acetonitrile. The gradient was performed as follows: 0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A with a flow rate of 400 µl/min.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"Water''s Xbridge amide (100 x 3mm, 3.5 um)","SOLVENT_A":"95% water/5% acetonitrile; 20 mM ammonium hydroxide; 20 mM ammonium acetate (pH 9.0)","SOLVENT_B":"100% acetonitrile","FLOW_GRADIENT":"0 min, 15% A; 2.5 min, 30% A; 7 min, 43% A; 16 min, 62% A; 16.1 to 18 min, 75% A; and 18 to 25 min, 15% A","FLOW_RATE":"400 μl/min","COLUMN_TEMPERATURE":"-"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"The capillary of the electrospray ionization source was set to 275°C, with sheath gas at 45 arbitrary units, auxiliary gas at 5 arbitrary units, and the spray voltage at 4.0 kV. In positive/negative polarity switching mode, a mass/charge ratio (m/z) scan range from 70 to 850 was chosen and MS1 data were collected at a resolution of 70,000. The automatic gain control target was set at 1 × 106, and the maximum injection time was 200 ms. The top five precursor ions were subsequently fragmented, in a data-dependent manner, using the higher-energy collisional dissociation cell set to 30% normalized collision energy in MS2 at a resolution power of 17,500."},

"MS_METABOLITE_DATA":{
"Units":"Peak area",

"Data":[{"Metabolite":"1-methylnicotinamide+","Les-Jas-20230207-01":"7799753.722","Les-Jas-20230207-02":"9988125.805","Les-Jas-20230207-03":"101245686","Les-Jas-20230207-04":"5881691.603","Les-Jas-20230207-05":"11988434.3","Les-Jas-20230207-06":"10567108.93","Les-Jas-20230207-07":"9463413.658","Les-Jas-20230207-08":"12827150.42","Les-Jas-20230207-09":"12740823.58","Les-Jas-20230207-10":"6582861.107","Les-Jas-20230207-11":"8340249.952","Les-Jas-20230207-12":"9998279.773"},{"Metabolite":"1-phenylethylamine","Les-Jas-20230207-01":"26120857.13","Les-Jas-20230207-02":"41558361.92","Les-Jas-20230207-03":"20354189.53","Les-Jas-20230207-04":"24185851.29","Les-Jas-20230207-05":"18321220.64","Les-Jas-20230207-06":"21350833.69","Les-Jas-20230207-07":"23600905.79","Les-Jas-20230207-08":"15889503.15","Les-Jas-20230207-09":"21276939","Les-Jas-20230207-10":"14912257.24","Les-Jas-20230207-11":"19721635.82","Les-Jas-20230207-12":"11258938.66"},{"Metabolite":"2- / 3-phosphoglycerate","Les-Jas-20230207-01":"84949.89334","Les-Jas-20230207-02":"12877.84771","Les-Jas-20230207-03":"55689.24041","Les-Jas-20230207-04":"93153.52321","Les-Jas-20230207-05":"208778.4263","Les-Jas-20230207-06":"94520.66309","Les-Jas-20230207-07":"166466.4498","Les-Jas-20230207-08":"110833.5754","Les-Jas-20230207-09":"195366.2652","Les-Jas-20230207-10":"30983.77452","Les-Jas-20230207-11":"136049.0119","Les-Jas-20230207-12":"204319.8309"},{"Metabolite":"2,3-bisphospho-D-glycerate","Les-Jas-20230207-01":"19056.06926","Les-Jas-20230207-02":"29405.57087","Les-Jas-20230207-03":"42057.0934","Les-Jas-20230207-04":"13487.16913","Les-Jas-20230207-05":"38106.73347","Les-Jas-20230207-06":"18097.26655","Les-Jas-20230207-07":"49880.91058","Les-Jas-20230207-08":"31672.6562","Les-Jas-20230207-09":"197296.9951","Les-Jas-20230207-10":"0","Les-Jas-20230207-11":"0","Les-Jas-20230207-12":"23575.15544"},{"Metabolite":"2-/ 3- hydroxybutanoic acid","Les-Jas-20230207-01":"1990688.36","Les-Jas-20230207-02":"2399851.501","Les-Jas-20230207-03":"3661050.253","Les-Jas-20230207-04":"2572966.962","Les-Jas-20230207-05":"2196157.852","Les-Jas-20230207-06":"1889991.919","Les-Jas-20230207-07":"1516528.457","Les-Jas-20230207-08":"4792085.654","Les-Jas-20230207-09":"1628263.441","Les-Jas-20230207-10":"857144.0701","Les-Jas-20230207-11":"988712.9176","Les-Jas-20230207-12":"1278688.933"},{"Metabolite":"2-aminobutyric / 2-/3-aminoisobutyric acid","Les-Jas-20230207-01":"10855348.63","Les-Jas-20230207-02":"10312868.05","Les-Jas-20230207-03":"38023857.95","Les-Jas-20230207-04":"5982741.669","Les-Jas-20230207-05":"19529378.12","Les-Jas-20230207-06":"23090850.66","Les-Jas-20230207-07":"9095741.972","Les-Jas-20230207-08":"10998188.66","Les-Jas-20230207-09":"10740091.61","Les-Jas-20230207-10":"9968958.188","Les-Jas-20230207-11":"12001992.4","Les-Jas-20230207-12":"14270815.84"},{"Metabolite":"2-HG","Les-Jas-20230207-01":"699900.6745","Les-Jas-20230207-02":"421520.1022","Les-Jas-20230207-03":"2128966.604","Les-Jas-20230207-04":"2171668.009","Les-Jas-20230207-05":"1436197.529","Les-Jas-20230207-06":"1207052.651","Les-Jas-20230207-07":"1404211.686","Les-Jas-20230207-08":"516098.2545","Les-Jas-20230207-09":"632259.7387","Les-Jas-20230207-10":"1729399.521","Les-Jas-20230207-11":"2668943.245","Les-Jas-20230207-12":"1061870.248"},{"Metabolite":"2-ketovaleric acid","Les-Jas-20230207-01":"2923653.594","Les-Jas-20230207-02":"4804277.216","Les-Jas-20230207-03":"4131086.107","Les-Jas-20230207-04":"0","Les-Jas-20230207-05":"4448561.063","Les-Jas-20230207-06":"0","Les-Jas-20230207-07":"0","Les-Jas-20230207-08":"0","Les-Jas-20230207-09":"4004488.449","Les-Jas-20230207-10":"2681236.576","Les-Jas-20230207-11":"0","Les-Jas-20230207-12":"1434835.284"},{"Metabolite":"3- / 4-hydroxyphenylacetic acid","Les-Jas-20230207-01":"172996.7616","Les-Jas-20230207-02":"410045.9231","Les-Jas-20230207-03":"525886.8666","Les-Jas-20230207-04":"335619.2159","Les-Jas-20230207-05":"437487.7642","Les-Jas-20230207-06":"115848.6473","Les-Jas-20230207-07":"77095.52924","Les-Jas-20230207-08":"194161.7969","Les-Jas-20230207-09":"189288.8752","Les-Jas-20230207-10":"438836.5616","Les-Jas-20230207-11":"211274.9726","Les-Jas-20230207-12":"299449.8063"},{"Metabolite":"3- / 7-methylguanine","Les-Jas-20230207-01":"74899.26218","Les-Jas-20230207-02":"186620.5665","Les-Jas-20230207-03":"137118.5099","Les-Jas-20230207-04":"121951.4192","Les-Jas-20230207-05":"89176.78877","Les-Jas-20230207-06":"73211.48587","Les-Jas-20230207-07":"72589.93965","Les-Jas-20230207-08":"84399.15644","Les-Jas-20230207-09":"111363.3631","Les-Jas-20230207-10":"82452.95079","Les-Jas-20230207-11":"69990.45495","Les-Jas-20230207-12":"100718.0186"},{"Metabolite":"3-phospho-serine","Les-Jas-20230207-01":"27434.84259","Les-Jas-20230207-02":"15888.37665","Les-Jas-20230207-03":"46514.42233","Les-Jas-20230207-04":"0","Les-Jas-20230207-05":"36430.27987","Les-Jas-20230207-06":"76169.75074","Les-Jas-20230207-07":"0","Les-Jas-20230207-08":"10129.46044","Les-Jas-20230207-09":"67075.80272","Les-Jas-20230207-10":"0","Les-Jas-20230207-11":"0","Les-Jas-20230207-12":"0"},{"Metabolite":"4-hydroxybutyric acid","Les-Jas-20230207-01":"63462349.93","Les-Jas-20230207-02":"74428859.53","Les-Jas-20230207-03":"63564903.35","Les-Jas-20230207-04":"53276796.14","Les-Jas-20230207-05":"73926349.18","Les-Jas-20230207-06":"57914119.82","Les-Jas-20230207-07":"72033043.14","Les-Jas-20230207-08":"62581624.05","Les-Jas-20230207-09":"64401029.05","Les-Jas-20230207-10":"63377539.29","Les-Jas-20230207-11":"70312065.03","Les-Jas-20230207-12":"61829598.63"},{"Metabolite":"4-hydroxyproline / 5-aminolevulinate","Les-Jas-20230207-01":"1267747.265","Les-Jas-20230207-02":"2299814.746","Les-Jas-20230207-03":"2468877.447","Les-Jas-20230207-04":"3814865.906","Les-Jas-20230207-05":"1436432.107","Les-Jas-20230207-06":"2631991.738","Les-Jas-20230207-07":"3765681.451","Les-Jas-20230207-08":"4991645.363","Les-Jas-20230207-09":"3533753.123","Les-Jas-20230207-10":"4620077.039","Les-Jas-20230207-11":"2766671.365","Les-Jas-20230207-12":"1737941.529"},{"Metabolite":"5-HIAA","Les-Jas-20230207-01":"16582.2178","Les-Jas-20230207-02":"23005.90963","Les-Jas-20230207-03":"0","Les-Jas-20230207-04":"16333.24929","Les-Jas-20230207-05":"0","Les-Jas-20230207-06":"15231.30022","Les-Jas-20230207-07":"25860.20823","Les-Jas-20230207-08":"49959.56857","Les-Jas-20230207-09":"55075.57132","Les-Jas-20230207-10":"66430.49582","Les-Jas-20230207-11":"32238.58004","Les-Jas-20230207-12":"0"},{"Metabolite":"5-methylcytosine","Les-Jas-20230207-01":"427256.0514","Les-Jas-20230207-02":"510192.1725","Les-Jas-20230207-03":"278309.8988","Les-Jas-20230207-04":"497999.3672","Les-Jas-20230207-05":"356186.3373","Les-Jas-20230207-06":"261762.6216","Les-Jas-20230207-07":"416026.6246","Les-Jas-20230207-08":"230263.1075","Les-Jas-20230207-09":"407365.4994","Les-Jas-20230207-10":"418693.816","Les-Jas-20230207-11":"197013.4013","Les-Jas-20230207-12":"351058.027"},{"Metabolite":"5''-methylthioadenosine","Les-Jas-20230207-01":"8511086.144","Les-Jas-20230207-02":"9924309.628","Les-Jas-20230207-03":"11842653.11","Les-Jas-20230207-04":"14058280.93","Les-Jas-20230207-05":"9724246.007","Les-Jas-20230207-06":"13322407.22","Les-Jas-20230207-07":"13755550.25","Les-Jas-20230207-08":"12684353.15","Les-Jas-20230207-09":"16800297.76","Les-Jas-20230207-10":"30040716.67","Les-Jas-20230207-11":"15249912.61","Les-Jas-20230207-12":"27092794.78"},{"Metabolite":"6-phosphogluconic acid","Les-Jas-20230207-01":"1085867.213","Les-Jas-20230207-02":"1640464.52","Les-Jas-20230207-03":"2207001.608","Les-Jas-20230207-04":"2615904.315","Les-Jas-20230207-05":"3218794.027","Les-Jas-20230207-06":"4075907.128","Les-Jas-20230207-07":"10311693.9","Les-Jas-20230207-08":"10296985.58","Les-Jas-20230207-09":"15834232.62","Les-Jas-20230207-10":"26361539.38","Les-Jas-20230207-11":"20948047.72","Les-Jas-20230207-12":"30124456.82"},{"Metabolite":"acetylcarnitine+","Les-Jas-20230207-01":"48566803.84","Les-Jas-20230207-02":"18363645.52","Les-Jas-20230207-03":"212876291","Les-Jas-20230207-04":"19837473.54","Les-Jas-20230207-05":"287069457.3","Les-Jas-20230207-06":"179279984.4","Les-Jas-20230207-07":"75981677.23","Les-Jas-20230207-08":"68692630.65","Les-Jas-20230207-09":"64065401","Les-Jas-20230207-10":"19640895.77","Les-Jas-20230207-11":"124642770.2","Les-Jas-20230207-12":"114166825.7"},{"Metabolite":"acetylcholine+","Les-Jas-20230207-01":"11991841.05","Les-Jas-20230207-02":"7988744.884","Les-Jas-20230207-03":"9087320.626","Les-Jas-20230207-04":"9100760.936","Les-Jas-20230207-05":"11366430.62","Les-Jas-20230207-06":"8688015.254","Les-Jas-20230207-07":"6007836.129","Les-Jas-20230207-08":"5327051.955","Les-Jas-20230207-09":"7524023.019","Les-Jas-20230207-10":"5459859.96","Les-Jas-20230207-11":"4745009.824","Les-Jas-20230207-12":"5434795.357"},{"Metabolite":"acrylic acid","Les-Jas-20230207-01":"19669813.64","Les-Jas-20230207-02":"22941468.84","Les-Jas-20230207-03":"23382310.17","Les-Jas-20230207-04":"12261121.63","Les-Jas-20230207-05":"14580505.65","Les-Jas-20230207-06":"18835140.02","Les-Jas-20230207-07":"22738420.88","Les-Jas-20230207-08":"18425950.66","Les-Jas-20230207-09":"17357829.27","Les-Jas-20230207-10":"16991668.52","Les-Jas-20230207-11":"22046074.99","Les-Jas-20230207-12":"18719627.44"},{"Metabolite":"adenine","Les-Jas-20230207-01":"4294200.079","Les-Jas-20230207-02":"5966277.482","Les-Jas-20230207-03":"3639251.029","Les-Jas-20230207-04":"6084538.399","Les-Jas-20230207-05":"8352801.529","Les-Jas-20230207-06":"6056739.93","Les-Jas-20230207-07":"5331012.811","Les-Jas-20230207-08":"4709060.691","Les-Jas-20230207-09":"5346508.183","Les-Jas-20230207-10":"4809310.443","Les-Jas-20230207-11":"3658842.261","Les-Jas-20230207-12":"6095600.471"},{"Metabolite":"adenosine","Les-Jas-20230207-01":"22646507.56","Les-Jas-20230207-02":"14804891.8","Les-Jas-20230207-03":"12998135.21","Les-Jas-20230207-04":"6966701.883","Les-Jas-20230207-05":"6337404.156","Les-Jas-20230207-06":"14903123.96","Les-Jas-20230207-07":"13330824.54","Les-Jas-20230207-08":"7855549.022","Les-Jas-20230207-09":"15910413.14","Les-Jas-20230207-10":"4486852.649","Les-Jas-20230207-11":"7709675.566","Les-Jas-20230207-12":"7213814.877"},{"Metabolite":"ADP / 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