{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002467","ANALYSIS_ID":"AN004023","VERSION":"1","CREATED_ON":"February 7, 2023, 8:36 am"},

"PROJECT":{"PROJECT_TITLE":"Nano-hijacked myeloid cells potentiate antitumor immunity and radiotherapy for glioblastoma","PROJECT_TYPE":"LC-MS/MS","PROJECT_SUMMARY":"Abstract: Radiation therapy is a key component of the standard of care for glioblastoma (GBM). Although this treatment is known to trigger pro-inflammatory immune responses, it also results in several immune resistance mechanisms such as the upregulation of CD47 by tumors leading to avoidance of phagocytosis and the overexpression of PD-L1 in tumor-associated myeloid cells (TAMCs). Leveraging these RT-elicited processes, we generated a bispecific-lipid nanoparticle (B-LNP) that engaged TAMCs to glioma cells via anti-CD47/PD-L1 dual-ligation. We show that B-LNP blocked these two vital immune checkpoint molecules and promoted the phagocytic activity of TAMCs. In order to boost subsequent T cell recruitment and antitumor activity after tumor engulfment, the B-LNP was encapsulated with diABZI, a non-nucleotidyl agonist for stimulator of interferon genes (STING). In vivo treatment with the diABZI-loaded B-LNP induced a transcriptomic and metabolic switch in TAMCs, transforming them into potent antitumor effector cells, which induced T cell infiltration and activation of in the brain tumors. In preclinical murine glioma models, B-LNP therapy significantly potentiated the antitumor effects of radiotherapy, promoted brain tumor regression, and induced immunological memory against gliomas. The nano37 therapy was efficacious through both intra-tumoral and systemic delivery routes. In summary, our study shows a unique nanotechnology-based approach that hijacks multiple immune checkpoints to boost potent and long-lasting antitumor immunity against GBM.","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":"Nano-hijacked myeloid cells potentiate antitumor immunity and radiotherapy for glioblastoma","STUDY_TYPE":"IR versus IR + LNP","STUDY_SUMMARY":"Abstract: Radiation therapy is a key component of the standard of care for glioblastoma (GBM). Although this treatment is known to trigger pro-inflammatory immune responses, it also results in several immune resistance mechanisms such as the upregulation of CD47 by tumors leading to avoidance of phagocytosis and the overexpression of PD-L1 in tumor-associated myeloid cells (TAMCs). Leveraging these RT-elicited processes, we generated a bispecific-lipid nanoparticle (B-LNP) that engaged TAMCs to glioma cells via anti-CD47/PD-L1 dual-ligation. We show that B-LNP blocked these two vital immune checkpoint molecules and promoted the phagocytic activity of TAMCs. In order to boost subsequent T cell recruitment and antitumor activity after tumor engulfment, the B-LNP was encapsulated with diABZI, a non-nucleotidyl agonist for stimulator of interferon genes (STING). In vivo treatment with the diABZI-loaded B-LNP induced a transcriptomic and metabolic switch in TAMCs, transforming them into potent antitumor effector cells, which induced T cell infiltration and activation of in the brain tumors. In preclinical murine glioma models, B-LNP therapy significantly potentiated the antitumor effects of radiotherapy, promoted brain tumor regression, and induced immunological memory against gliomas. The nano37 therapy was efficacious through both intra-tumoral and systemic delivery routes. In summary, our study shows a unique nanotechnology-based approach that hijacks multiple immune checkpoints to boost potent and long-lasting antitumor immunity against GBM.","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":"6"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","TAXONOMY_ID":"10090"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-01",
"Factors":{"Treatment":"IR"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-01"}
},
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-02",
"Factors":{"Treatment":"IR"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-02"}
},
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-03",
"Factors":{"Treatment":"IR"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-03"}
},
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-04",
"Factors":{"Treatment":"IR + LNP"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-04"}
},
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-05",
"Factors":{"Treatment":"IR + LNP"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-05"}
},
{
"Subject ID":"-",
"Sample ID":"Zha-Jas-20210625-06",
"Factors":{"Treatment":"IR + LNP"},
"Additional sample data":{"RAW_FILE_NAME":"Zha-Jas-20210625-06"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Magnetic bead isolation of cells To isolate specific populations of cells, single-cell suspensions as isolated above are preblocked with anti-CD16/32 for 15 min at 4°C. We then used the biotinylated anti-Gr1 (clone RB6-8C5) (all from Thermo Fisher Scientific) to label murine myeloid cells. Next, the cells were washed and then incubated with anti-biotin magnetic beads (Miltenyi Biotec) before performing manual positive selection using MS columns (Miltenyi Biotec). Purified cells were analyzed for all downstream metabolic analyses.","SAMPLE_TYPE":"Brain"},

"TREATMENT":{"TREATMENT_SUMMARY":"GR1 cells were isolated from mice after 9Gy Radiation or 9Gy radiation and nanoparticle therapy"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Isolated TAMC and CD8+ T cells samples were dried using a SpeedVac. Acetonitrile (50%) was added to the tube for reconstitution following overtaxing for 30 s. Sample solution was then centrifuged for 15 min at 20,000g and 4°C. Supernatant was collected for LC-MS analysis. 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. 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."},

"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).","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Q Exactive™ Plus Hybrid Quadrupole-Orbitrap™ Mass Spectrometer","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":"275"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Plus Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","MS_COMMENTS":"Isolated TAMC and CD8+ T cells samples were dried using a SpeedVac. Acetonitrile (50%) was added to the tube for reconstitution following overtaxing for 30 s. Sample solution was then centrifuged for 15 min at 20,000g and 4°C. Supernatant was collected for LC-MS analysis. 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. 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":"Normalized peak area",

"Data":[{"Metabolite":"(S)-4-hydroxymandelonitrile","Zha-Jas-20210625-01":"272476","Zha-Jas-20210625-02":"187098","Zha-Jas-20210625-03":"148565","Zha-Jas-20210625-04":"81421","Zha-Jas-20210625-05":"712900","Zha-Jas-20210625-06":"434657"},{"Metabolite":"1,3-bisphospho-D-glycerate","Zha-Jas-20210625-01":"6263917","Zha-Jas-20210625-02":"9058076","Zha-Jas-20210625-03":"8058918","Zha-Jas-20210625-04":"2069914","Zha-Jas-20210625-05":"13117887","Zha-Jas-20210625-06":"3788895"},{"Metabolite":"1-methylnicotinamide+","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"2792617","Zha-Jas-20210625-05":"17617170","Zha-Jas-20210625-06":"14958269"},{"Metabolite":"1-phenylethylamine","Zha-Jas-20210625-01":"6454873","Zha-Jas-20210625-02":"3587045","Zha-Jas-20210625-03":"2617278","Zha-Jas-20210625-04":"2786468","Zha-Jas-20210625-05":"3368768","Zha-Jas-20210625-06":"2377033"},{"Metabolite":"2- / 3-phosphoglycerate","Zha-Jas-20210625-01":"38736246","Zha-Jas-20210625-02":"44890450","Zha-Jas-20210625-03":"34332090","Zha-Jas-20210625-04":"44579378","Zha-Jas-20210625-05":"62659596","Zha-Jas-20210625-06":"39837433"},{"Metabolite":"2-(Formamido)-N1-(5-phospho-D-ribosyl)acetamidine","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"2,3-bisphospho-D-glycerate","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"2-aminobutanoate / 2-aminoisobutanoate","Zha-Jas-20210625-01":"14566490","Zha-Jas-20210625-02":"71386144","Zha-Jas-20210625-03":"12820411","Zha-Jas-20210625-04":"9739895","Zha-Jas-20210625-05":"9614177","Zha-Jas-20210625-06":"10110497"},{"Metabolite":"2-HG","Zha-Jas-20210625-01":"1707088","Zha-Jas-20210625-02":"498898","Zha-Jas-20210625-03":"830454","Zha-Jas-20210625-04":"786590","Zha-Jas-20210625-05":"441963","Zha-Jas-20210625-06":"264808"},{"Metabolite":"2-hydroxybutanoic acid","Zha-Jas-20210625-01":"1830653","Zha-Jas-20210625-02":"2283394","Zha-Jas-20210625-03":"1820562","Zha-Jas-20210625-04":"2403435","Zha-Jas-20210625-05":"2284568","Zha-Jas-20210625-06":"2109845"},{"Metabolite":"2-keto-isovaleric acid","Zha-Jas-20210625-01":"3322779","Zha-Jas-20210625-02":"6276340","Zha-Jas-20210625-03":"5691331","Zha-Jas-20210625-04":"8134843","Zha-Jas-20210625-05":"8533267","Zha-Jas-20210625-06":"5697467"},{"Metabolite":"2-ketovaleric acid","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"2-oxoadipic acid","Zha-Jas-20210625-01":"299991","Zha-Jas-20210625-02":"311175","Zha-Jas-20210625-03":"465596","Zha-Jas-20210625-04":"384353","Zha-Jas-20210625-05":"186531","Zha-Jas-20210625-06":"0"},{"Metabolite":"2-oxobutanoic acid","Zha-Jas-20210625-01":"9384638","Zha-Jas-20210625-02":"23157730","Zha-Jas-20210625-03":"21338340","Zha-Jas-20210625-04":"25486763","Zha-Jas-20210625-05":"56136063","Zha-Jas-20210625-06":"55495926"},{"Metabolite":"3- / 4 -hydroxybutanoic acid","Zha-Jas-20210625-01":"47943599","Zha-Jas-20210625-02":"42574046","Zha-Jas-20210625-03":"37179110","Zha-Jas-20210625-04":"48796756","Zha-Jas-20210625-05":"54360462","Zha-Jas-20210625-06":"65489890"},{"Metabolite":"3- / 4-hydroxyphenylacetic acid / vanillin /","Zha-Jas-20210625-01":"6836661","Zha-Jas-20210625-02":"6678657","Zha-Jas-20210625-03":"7072598","Zha-Jas-20210625-04":"5876842","Zha-Jas-20210625-05":"5150301","Zha-Jas-20210625-06":"4562461"},{"Metabolite":"3- / 7-methylguanine","Zha-Jas-20210625-01":"471722","Zha-Jas-20210625-02":"454629","Zha-Jas-20210625-03":"313972","Zha-Jas-20210625-04":"52868","Zha-Jas-20210625-05":"182320","Zha-Jas-20210625-06":"76663"},{"Metabolite":"3,4-dihydroxymandelic acid","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3,4-dihydroxyphenylpropanoate","Zha-Jas-20210625-01":"12903","Zha-Jas-20210625-02":"27292","Zha-Jas-20210625-03":"24054","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"15341","Zha-Jas-20210625-06":"11280"},{"Metabolite":"3-aminoisobutanoate","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3-hydroxy-anthranilic acid","Zha-Jas-20210625-01":"494582","Zha-Jas-20210625-02":"203194","Zha-Jas-20210625-03":"249458","Zha-Jas-20210625-04":"206035","Zha-Jas-20210625-05":"180329","Zha-Jas-20210625-06":"248458"},{"Metabolite":"3-hydroxykynurenine","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3-hydroxypropanoic acid","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3-methoxytyramine","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3-phosphohydroxypyruvic acid","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"185362","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"3-phospho-serine","Zha-Jas-20210625-01":"8286866","Zha-Jas-20210625-02":"7985580","Zha-Jas-20210625-03":"8815299","Zha-Jas-20210625-04":"13502454","Zha-Jas-20210625-05":"12587523","Zha-Jas-20210625-06":"9999077"},{"Metabolite":"3-ureidopropionic acid","Zha-Jas-20210625-01":"429041","Zha-Jas-20210625-02":"69209","Zha-Jas-20210625-03":"122092","Zha-Jas-20210625-04":"465426","Zha-Jas-20210625-05":"1444790","Zha-Jas-20210625-06":"1076560"},{"Metabolite":"4-hydroxyphenylpyruvic acid","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"24519","Zha-Jas-20210625-06":"0"},{"Metabolite":"4-hydroxyproline / 5-aminolevulinate / N-acetyl-b-alanine","Zha-Jas-20210625-01":"14254238","Zha-Jas-20210625-02":"13186677","Zha-Jas-20210625-03":"16235269","Zha-Jas-20210625-04":"4219744","Zha-Jas-20210625-05":"2448221","Zha-Jas-20210625-06":"1402869"},{"Metabolite":"4-phosphopantothenate","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"4-trimethylammoniobutanoate+","Zha-Jas-20210625-01":"15093882","Zha-Jas-20210625-02":"9034312","Zha-Jas-20210625-03":"9632315","Zha-Jas-20210625-04":"6042373","Zha-Jas-20210625-05":"5213319","Zha-Jas-20210625-06":"2689892"},{"Metabolite":"5,6-dihydroxyindole","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"5-HIAA","Zha-Jas-20210625-01":"15938","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"10417","Zha-Jas-20210625-04":"28848","Zha-Jas-20210625-05":"9876","Zha-Jas-20210625-06":"0"},{"Metabolite":"5-methylcytosine","Zha-Jas-20210625-01":"2428204","Zha-Jas-20210625-02":"1286670","Zha-Jas-20210625-03":"727951","Zha-Jas-20210625-04":"1257153","Zha-Jas-20210625-05":"1571172","Zha-Jas-20210625-06":"746473"},{"Metabolite":"5-methyl-THF","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"5'-methylthioadenosine","Zha-Jas-20210625-01":"35416995","Zha-Jas-20210625-02":"33268361","Zha-Jas-20210625-03":"35520454","Zha-Jas-20210625-04":"25104051","Zha-Jas-20210625-05":"22827526","Zha-Jas-20210625-06":"16426689"},{"Metabolite":"6-aminocaproic acid","Zha-Jas-20210625-01":"6739697","Zha-Jas-20210625-02":"11738147","Zha-Jas-20210625-03":"23454759","Zha-Jas-20210625-04":"7165117","Zha-Jas-20210625-05":"12416428","Zha-Jas-20210625-06":"14426422"},{"Metabolite":"6-methylnicotinamide","Zha-Jas-20210625-01":"93918527","Zha-Jas-20210625-02":"110773023","Zha-Jas-20210625-03":"53501161","Zha-Jas-20210625-04":"22247906","Zha-Jas-20210625-05":"10000398","Zha-Jas-20210625-06":"8264686"},{"Metabolite":"6-O-methylguanine","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"6-phosphogluconic acid","Zha-Jas-20210625-01":"67139","Zha-Jas-20210625-02":"166112","Zha-Jas-20210625-03":"35807","Zha-Jas-20210625-04":"54529","Zha-Jas-20210625-05":"49439","Zha-Jas-20210625-06":"105135"},{"Metabolite":"6-phosphoglucono-D-lactone","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"acetoacetic acid / succinic semialdehyde","Zha-Jas-20210625-01":"29571699","Zha-Jas-20210625-02":"28678489","Zha-Jas-20210625-03":"26187208","Zha-Jas-20210625-04":"32078982","Zha-Jas-20210625-05":"16995021","Zha-Jas-20210625-06":"10661179"},{"Metabolite":"acetylcholine+","Zha-Jas-20210625-01":"84040616","Zha-Jas-20210625-02":"66960674","Zha-Jas-20210625-03":"59614939","Zha-Jas-20210625-04":"26383199","Zha-Jas-20210625-05":"11328997","Zha-Jas-20210625-06":"12039465"},{"Metabolite":"acetyl-CoA","Zha-Jas-20210625-01":"0","Zha-Jas-20210625-02":"0","Zha-Jas-20210625-03":"0","Zha-Jas-20210625-04":"0","Zha-Jas-20210625-05":"0","Zha-Jas-20210625-06":"0"},{"Metabolite":"aconitic 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"Metabolites":[{"Metabolite":"(S)-4-hydroxymandelonitrile","KEGG ID":"C03742"},{"Metabolite":"1,3-bisphospho-D-glycerate","KEGG ID":"C00236"},{"Metabolite":"1-methylnicotinamide+","KEGG ID":"C02918"},{"Metabolite":"1-phenylethylamine","KEGG ID":"C02455"},{"Metabolite":"2- / 3-phosphoglycerate","KEGG ID":"C00631 / C00197"},{"Metabolite":"2-(Formamido)-N1-(5-phospho-D-ribosyl)acetamidine","KEGG ID":"C04640"},{"Metabolite":"2,3-bisphospho-D-glycerate","KEGG ID":"C01159-C03339"},{"Metabolite":"2-aminobutanoate / 2-aminoisobutanoate","KEGG ID":"C02261 / C03665"},{"Metabolite":"2-HG","KEGG ID":"C02630"},{"Metabolite":"2-hydroxybutanoic acid","KEGG ID":"C05984"},{"Metabolite":"2-keto-isovaleric acid","KEGG ID":"C00141"},{"Metabolite":"2-ketovaleric acid","KEGG ID":"C06255"},{"Metabolite":"2-oxoadipic acid","KEGG ID":"C00322"},{"Metabolite":"2-oxobutanoic acid","KEGG ID":"C00109"},{"Metabolite":"3- / 4 -hydroxybutanoic acid","KEGG ID":"C01089 / C00989"},{"Metabolite":"3- / 4-hydroxyphenylacetic acid / vanillin /","KEGG ID":"C05593 / C00642 / C00755"},{"Metabolite":"3- / 7-methylguanine","KEGG ID":"C02230 / C02242"},{"Metabolite":"3,4-dihydroxymandelic acid","KEGG ID":"C05580"},{"Metabolite":"3,4-dihydroxyphenylpropanoate","KEGG ID":"C10447"},{"Metabolite":"3-aminoisobutanoate","KEGG ID":"C05145"},{"Metabolite":"3-hydroxy-anthranilic acid","KEGG ID":"C00632"},{"Metabolite":"3-hydroxykynurenine","KEGG ID":"C02794"},{"Metabolite":"3-hydroxypropanoic acid","KEGG ID":"C01013"},{"Metabolite":"3-methoxytyramine","KEGG ID":"C05587"},{"Metabolite":"3-phosphohydroxypyruvic acid","KEGG ID":"C03232"},{"Metabolite":"3-phospho-serine","KEGG ID":"C01005"},{"Metabolite":"3-ureidopropionic acid","KEGG ID":"C02642"},{"Metabolite":"4-hydroxyphenylpyruvic acid","KEGG ID":"C01179"},{"Metabolite":"4-hydroxyproline / 5-aminolevulinate / N-acetyl-b-alanine","KEGG ID":"C01157 / C00430 / C01073"},{"Metabolite":"4-phosphopantothenate","KEGG ID":"C03492"},{"Metabolite":"4-trimethylammoniobutanoate+","KEGG ID":"C01181"},{"Metabolite":"5,6-dihydroxyindole","KEGG ID":"C05578"},{"Metabolite":"5-HIAA","KEGG ID":"C05635"},{"Metabolite":"5-methylcytosine","KEGG ID":"C02376"},{"Metabolite":"5-methyl-THF","KEGG ID":"C00440"},{"Metabolite":"5'-methylthioadenosine","KEGG ID":"C00170"},{"Metabolite":"6-aminocaproic acid","KEGG ID":"C02378"},{"Metabolite":"6-methylnicotinamide","KEGG ID":"n/a"},{"Metabolite":"6-O-methylguanine","KEGG ID":"n/a"},{"Metabolite":"6-phosphogluconic acid","KEGG ID":"C00345"},{"Metabolite":"6-phosphoglucono-D-lactone","KEGG ID":"C01236"},{"Metabolite":"acetoacetic acid / succinic semialdehyde","KEGG ID":"C00164 / C00232"},{"Metabolite":"acetylcholine+","KEGG ID":"C01996"},{"Metabolite":"acetyl-CoA","KEGG ID":"C00024"},{"Metabolite":"aconitic acid","KEGG ID":"C00417-C02341"},{"Metabolite":"adenine","KEGG ID":"C00147"},{"Metabolite":"adenosine","KEGG ID":"C00212"},{"Metabolite":"adenylosuccinate","KEGG ID":"C03794"},{"Metabolite":"ADP / dGDP","KEGG ID":"C00008 / C00361"},{"Metabolite":"ADP-D-glucose","KEGG ID":"C00498"},{"Metabolite":"agmatine","KEGG ID":"C00179"},{"Metabolite":"AICA ribonucleotide","KEGG ID":"C04677"},{"Metabolite":"a-KG","KEGG ID":"C00026"},{"Metabolite":"alanine","KEGG ID":"C00041-C00133"},{"Metabolite":"alanine methyl ester","KEGG ID":"n/a"},{"Metabolite":"allantoin","KEGG ID":"C01551"},{"Metabolite":"AMP / dGMP","KEGG ID":"C00020 / C00362"},{"Metabolite":"anandamide","KEGG ID":"C11695"},{"Metabolite":"arginine","KEGG ID":"C02385"},{"Metabolite":"ascorbic acid","KEGG ID":"C00072"},{"Metabolite":"asparagine","KEGG ID":"C16438"},{"Metabolite":"aspartic acid","KEGG ID":"C16433"},{"Metabolite":"ATP / dGTP","KEGG ID":"C00002 / C00286"},{"Metabolite":"b-alanine","KEGG ID":"C00099"},{"Metabolite":"betaine","KEGG ID":"C00719"},{"Metabolite":"butyric acid","KEGG ID":"C00246"},{"Metabolite":"butyric acid(iso)","KEGG ID":"C02632"},{"Metabolite":"carbamoyl phosphate","KEGG ID":"C00169"},{"Metabolite":"carnitine+","KEGG ID":"C00487"},{"Metabolite":"CDP","KEGG ID":"C00112"},{"Metabolite":"CDP-choline","KEGG ID":"C00307"},{"Metabolite":"cGMP","KEGG ID":"C00943"},{"Metabolite":"choline+","KEGG ID":"C00114"},{"Metabolite":"citraconic acid","KEGG ID":"C02226"},{"Metabolite":"citramalic acid","KEGG ID":"C00815"},{"Metabolite":"citrate / citrate(iso)","KEGG ID":"C00158 / C00311"},{"Metabolite":"citrulline","KEGG ID":"C00327"},{"Metabolite":"CMP","KEGG ID":"C00055"},{"Metabolite":"cortisol","KEGG ID":"C00735"},{"Metabolite":"creatine","KEGG ID":"C00300"},{"Metabolite":"creatinine","KEGG ID":"C00791"},{"Metabolite":"CTP","KEGG ID":"C00063"},{"Metabolite":"cystathionine","KEGG ID":"C00542"},{"Metabolite":"cysteine","KEGG ID":"C00736-C00097"},{"Metabolite":"cysteine-S-sulfate","KEGG ID":"C05824"},{"Metabolite":"cystine","KEGG ID":"C01420"},{"Metabolite":"cytidine","KEGG ID":"C00475"},{"Metabolite":"cytosine","KEGG ID":"C00380"},{"Metabolite":"dADP","KEGG ID":"C00206"},{"Metabolite":"dAMP","KEGG ID":"C00360"},{"Metabolite":"D-arabinono-1,4-lactone","KEGG ID":"C00652"},{"Metabolite":"dATP","KEGG ID":"C00131"},{"Metabolite":"dCDP","KEGG ID":"C00705"},{"Metabolite":"dCMP","KEGG ID":"C00239"},{"Metabolite":"dCTP","KEGG ID":"C00458"},{"Metabolite":"dehydroascorbic acid","KEGG ID":"C05422"},{"Metabolite":"deoxyguanosine","KEGG ID":"C00330"},{"Metabolite":"deoxyinosine","KEGG ID":"C05512"},{"Metabolite":"deoxyuridine","KEGG ID":"C00526"},{"Metabolite":"D-erythrose-4-phosphate","KEGG ID":"C00279"},{"Metabolite":"D-galactonic acid / D-gluconic acid","KEGG ID":"C00880 / C00257"},{"Metabolite":"D-galactosamine/glucosamine-1/6-phosphate","KEGG ID":"C03783 / C00352 / C06377 / C06156"},{"Metabolite":"dihydroorotate","KEGG ID":"C00337"},{"Metabolite":"dimethylglycine","KEGG ID":"C01026"},{"Metabolite":"DL-DOPA","KEGG ID":"C00355"},{"Metabolite":"DOPAC","KEGG ID":"C01161"},{"Metabolite":"dopamine","KEGG ID":"C03758"},{"Metabolite":"D-ribose-1/5-phosphate / D-ribulose 5-phosphate","KEGG ID":"C00620 / C00117 / C00199"},{"Metabolite":"D-sedoheptulose-7-phosphate","KEGG ID":"C05382"},{"Metabolite":"dTMP","KEGG ID":"C00364"},{"Metabolite":"dTTP","KEGG ID":"C00459"},{"Metabolite":"dUMP","KEGG ID":"C00365"},{"Metabolite":"dUTP","KEGG ID":"C00460"},{"Metabolite":"epinephrine","KEGG ID":"C00788"},{"Metabolite":"ethyl acetate","KEGG ID":"C00849"},{"Metabolite":"F-1,6/2,6-DP","KEGG ID":"C00354 / C00665"},{"Metabolite":"F1P","KEGG ID":"C01094"},{"Metabolite":"F6P / G1P / G6P","KEGG ID":"C00085 / C00103 / C00092"},{"Metabolite":"FAD","KEGG ID":"C00016"},{"Metabolite":"FMN","KEGG ID":"C00061"},{"Metabolite":"folic acid","KEGG ID":"C00504"},{"Metabolite":"fructose / mannose","KEGG ID":"C00095 / C00159"},{"Metabolite":"fumaric acid","KEGG ID":"C00122"},{"Metabolite":"GABA","KEGG ID":"C00334"},{"Metabolite":"galactosamine / glucosamine","KEGG ID":"C02262 / C00329"},{"Metabolite":"galactose / glucose","KEGG ID":"C00124 / C00267"},{"Metabolite":"GalNAC/GlcNAC/ManNAC","KEGG ID":"C01074 / C00140 / C00645"},{"Metabolite":"GAP / DHAP","KEGG ID":"C00118 / C00111"},{"Metabolite":"GDP","KEGG ID":"C00035"},{"Metabolite":"GDP-L-fucose","KEGG ID":"C00325"},{"Metabolite":"glucono-lactone","KEGG ID":"C00198"},{"Metabolite":"glutaconic acid","KEGG ID":"C02214"},{"Metabolite":"glutamine","KEGG ID":"C00303"},{"Metabolite":"glutaminic acid","KEGG ID":"C00302"},{"Metabolite":"glutathione","KEGG ID":"C00051"},{"Metabolite":"glutathione disulfide","KEGG ID":"C00127"},{"Metabolite":"glyceraldehyde","KEGG ID":"C02426"},{"Metabolite":"glyceric acid","KEGG ID":"C00258"},{"Metabolite":"glycerol 2/3-phosphate","KEGG ID":"C02979 / C00093"},{"Metabolite":"glycine","KEGG ID":"C00037"},{"Metabolite":"glycylglycine","KEGG ID":"C02037"},{"Metabolite":"GMP","KEGG ID":"C00144"},{"Metabolite":"GTP","KEGG ID":"C00044"},{"Metabolite":"guanidine","KEGG ID":"C17349"},{"Metabolite":"guanidinoacetate","KEGG 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ID":"C01432"},{"Metabolite":"L-arginino-succinate","KEGG ID":"C03406"},{"Metabolite":"L-cysteic acid","KEGG ID":"C00506"},{"Metabolite":"leucine / leucine(iso)","KEGG ID":"C16439 / C16434"},{"Metabolite":"levulinic acid","KEGG ID":"n/a"},{"Metabolite":"L-NMMA","KEGG ID":"C03884"},{"Metabolite":"L-tyrosine methyl ester","KEGG ID":"C03404"},{"Metabolite":"lysine","KEGG ID":"C00047/C00739"},{"Metabolite":"maleic acid","KEGG ID":"C01384"},{"Metabolite":"malic acid","KEGG ID":"C00711"},{"Metabolite":"malonyl-CoA","KEGG ID":"C00083"},{"Metabolite":"mesaconic acid","KEGG ID":"C01732"},{"Metabolite":"methionine","KEGG ID":"C00073"},{"Metabolite":"methyl acetoacetate","KEGG ID":"n/a"},{"Metabolite":"methylglyoxal","KEGG ID":"C00546"},{"Metabolite":"methylmalonic acid","KEGG ID":"C02170"},{"Metabolite":"mevalonic acid","KEGG ID":"C00418"},{"Metabolite":"N1/N8-acetylspermidine","KEGG ID":"C00612 / C01029"},{"Metabolite":"N-acetylaspartic acid","KEGG ID":"C01042"},{"Metabolite":"N-acetylaspartylglutamic acid","KEGG ID":"C12270"},{"Metabolite":"N-acetyl-DL-alanine","KEGG ID":"n/a"},{"Metabolite":"N-acetyl-DL-serine","KEGG ID":"n/a"},{"Metabolite":"N-acetyl-dopamine","KEGG ID":"n/a"},{"Metabolite":"N-acetylglucosamine 1/6-phosphate","KEGG ID":"C04501 / C00357"},{"Metabolite":"N-acetyl-L-glutamic acid","KEGG ID":"C00624"},{"Metabolite":"NAD+","KEGG ID":"C00003"},{"Metabolite":"NADH","KEGG ID":"C00004"},{"Metabolite":"NADP+","KEGG ID":"C00006"},{"Metabolite":"NADPH","KEGG ID":"C00005"},{"Metabolite":"N-carbamoyl-L-aspartate","KEGG ID":"C00438"},{"Metabolite":"nebularine","KEGG ID":"C01736"},{"Metabolite":"nicotinamide","KEGG ID":"C00153"},{"Metabolite":"nicotinamide riboside+","KEGG ID":"C03150"},{"Metabolite":"nicotinic acid","KEGG ID":"C00253"},{"Metabolite":"N-methyltryptamine","KEGG ID":"C06213"},{"Metabolite":"NMN","KEGG ID":"C00455"},{"Metabolite":"norepinephrine","KEGG ID":"C00547"},{"Metabolite":"O-acetyl-L-serine","KEGG ID":"C00979"},{"Metabolite":"octopamine","KEGG ID":"C04227"},{"Metabolite":"ornithine","KEGG ID":"C01602"},{"Metabolite":"orotic acid","KEGG ID":"C00295"},{"Metabolite":"orotidylic acid","KEGG ID":"C01103"},{"Metabolite":"oxalacetic acid","KEGG ID":"C00036"},{"Metabolite":"pantothenic acid","KEGG ID":"C00864"},{"Metabolite":"phenethylamine","KEGG ID":"C05332"},{"Metabolite":"phenylalanine","KEGG ID":"C02057"},{"Metabolite":"phenylephrine / synephrine","KEGG ID":"C07441 / C04548"},{"Metabolite":"phosphocreatine","KEGG ID":"C02305"},{"Metabolite":"phosphoenolpyruvate","KEGG ID":"C00074"},{"Metabolite":"phosphorylcholine+","KEGG ID":"C00588"},{"Metabolite":"picolinic acid","KEGG ID":"C10164"},{"Metabolite":"proline","KEGG ID":"C16435"},{"Metabolite":"propionic acid","KEGG ID":"C00163"},{"Metabolite":"PRPP","KEGG ID":"C00119"},{"Metabolite":"pseudouridine","KEGG ID":"C02067"},{"Metabolite":"putrescine","KEGG ID":"C00134"},{"Metabolite":"pyridoxine","KEGG ID":"C00314"},{"Metabolite":"pyroglutamic acid","KEGG ID":"C01879-C02237"},{"Metabolite":"pyrroline hydroxycarboxylic acid","KEGG ID":"C04281"},{"Metabolite":"pyruvic acid","KEGG ID":"C00022"},{"Metabolite":"quinolinic acid","KEGG ID":"C03722"},{"Metabolite":"S-adenosyl-L-homocysteine","KEGG ID":"C00021"},{"Metabolite":"S-adenosyl-L-methionine+","KEGG ID":"C00019"},{"Metabolite":"S-adenosylmethioninamine+","KEGG ID":"C01137"},{"Metabolite":"SAICAR","KEGG ID":"C04823"},{"Metabolite":"sarcosine","KEGG ID":"C00213"},{"Metabolite":"serine","KEGG ID":"C00716"},{"Metabolite":"serotonin","KEGG ID":"C00780"},{"Metabolite":"sorbitol / galactitol / L-iditol / manitol","KEGG ID":"C00794 / C01697 / C01507 / C00392"},{"Metabolite":"spermidine","KEGG ID":"C00315"},{"Metabolite":"spermine","KEGG ID":"C00750"},{"Metabolite":"succinic acid","KEGG ID":"C00042"},{"Metabolite":"succinyl-CoA","KEGG ID":"C00091"},{"Metabolite":"taurine","KEGG ID":"C00245"},{"Metabolite":"thiamine+","KEGG ID":"C00378"},{"Metabolite":"thymidine","KEGG ID":"C00214"},{"Metabolite":"thymine","KEGG ID":"C00178"},{"Metabolite":"tryptamine","KEGG ID":"C00398"},{"Metabolite":"tryptophan","KEGG ID":"C00806"},{"Metabolite":"tyramine","KEGG ID":"C00483"},{"Metabolite":"tyrosine","KEGG ID":"C01536"},{"Metabolite":"UDP","KEGG ID":"C00015"},{"Metabolite":"UDP-D-glucose","KEGG ID":"C00029"},{"Metabolite":"UDP-N-acetyl-galactosamine/UDP-N-acetyl-glucosamine","KEGG ID":"C00203 / C00043"},{"Metabolite":"UMP","KEGG ID":"C00105"},{"Metabolite":"uracil","KEGG ID":"C00106"},{"Metabolite":"urea","KEGG ID":"C00086"},{"Metabolite":"uric acid","KEGG ID":"C00366"},{"Metabolite":"uridine","KEGG ID":"C00299"},{"Metabolite":"UTP","KEGG ID":"C00075"},{"Metabolite":"valine","KEGG ID":"C16436"},{"Metabolite":"xanthine","KEGG ID":"C00385"},{"Metabolite":"xanthosine","KEGG ID":"C01762"},{"Metabolite":"xanthurenic acid","KEGG ID":"C02470"},{"Metabolite":"XMP","KEGG ID":"C00655"},{"Metabolite":"y-glutamylcysteine","KEGG ID":"C00669"}]
}

}