#METABOLOMICS WORKBENCH gmcgivney_20250506_102226 DATATRACK_ID:5889 STUDY_ID:ST003926 ANALYSIS_ID:AN006449 PROJECT_ID:PR002459 VERSION 1 CREATED_ON 05-23-2025 #PROJECT PR:PROJECT_TITLE Somatic CRISPR tumorigenesis and multi-omic analysis reveal a pentose phosphate PR:PROJECT_TITLE pathway disruption vulnerability in MPNSTs PR:PROJECT_TYPE GC-and LC-MS PR:PROJECT_SUMMARY Malignant Peripheral Nerve Sheath Tumors (MPNSTs) are aggressive and PR:PROJECT_SUMMARY chemo-resistant sarcomas with poor survival rates. Loss of CDKN2A or P53 PR:PROJECT_SUMMARY following NF1 disruption is a key event in MPNST development. Here, we employed PR:PROJECT_SUMMARY CRISPR/Cas9 somatic tumorigenesis in mice to identify transcriptomic and PR:PROJECT_SUMMARY metabolomic features distinguishing CDKN2A- versus P53-deleted MPNSTs. PR:PROJECT_SUMMARY Convergent, multi-omic analyses revealed CDKN2A-deleted MPNSTs are especially PR:PROJECT_SUMMARY dependent upon the pentose phosphate pathway (PPP) and NADPH metabolism for PR:PROJECT_SUMMARY growth and viability. Disruption of glucose-6-phosphate dehydrogenase (G6PD), PR:PROJECT_SUMMARY the PPP rate-limiting enzyme, slowed CDKN2A-deleted MPNST growth and sensitized PR:PROJECT_SUMMARY MPNSTs to standard-of-care chemotherapy. Knockdown of the redox-regulated PR:PROJECT_SUMMARY transcription factor NRF2 slowed MPNST growth and decreased G6PD transcription. PR:PROJECT_SUMMARY Analysis of patient MPNSTs identified a NRF2 gene signature correlating with PR:PROJECT_SUMMARY tumor transformation. Furthermore, G6PD and NRF2 expression in pan-cancer TCGA PR:PROJECT_SUMMARY samples correlates with patient survival. This work identifies NRF2-PPP PR:PROJECT_SUMMARY dependency as a targetable vulnerability in these difficult to treat MPNSTs, PR:PROJECT_SUMMARY particularly in the NF1/CDKN2A-deleted majority. PR:INSTITUTE University of Iowa PR:DEPARTMENT Molecular Physiology and Biophysics PR:LABORATORY Eric Taylor Lab PR:LAST_NAME McGivney PR:FIRST_NAME Gavin PR:ADDRESS 3316 Pappajohn Biomedical Discovery Building, Iowa City, IA, 52242, USA PR:EMAIL gavin-mcgivney@uiowa.edu PR:PHONE 5158646586 PR:DOI http://dx.doi.org/10.21228/M8SG17 #STUDY ST:STUDY_TITLE Somatic CRISPR tumorigenesis and multi-omic analysis reveal a pentose phosphate ST:STUDY_TITLE pathway disruption vulnerability in Malignant Peripheral Nerve Sheath Tumors ST:STUDY_TITLE (MPNSTs) ST:STUDY_SUMMARY Malignant Peripheral Nerve Sheath Tumors (MPNSTs) are aggressive and ST:STUDY_SUMMARY chemo-resistant sarcomas with poor survival rates. Loss of tumor suppressors ST:STUDY_SUMMARY CDKN2A or TP53 following NF1 (tumor suppressor) disruption is a key event in ST:STUDY_SUMMARY MPNST development. Here, we employed CRISPR/Cas9 somatic tumorigenesis in mice ST:STUDY_SUMMARY to identify transcriptomic and metabolomic features distinguishing CDKN2A- ST:STUDY_SUMMARY versus P53-deleted MPNSTs. Convergent, multi-omic analyses revealed ST:STUDY_SUMMARY CDKN2A-deleted MPNSTs are especially dependent upon the pentose phosphate ST:STUDY_SUMMARY pathway (PPP) and NADPH metabolism for growth and viability. Disruption of ST:STUDY_SUMMARY glucose-6-phosphate dehydrogenase (G6PD), the PPP rate-limiting enzyme, slowed ST:STUDY_SUMMARY CDKN2A-deleted MPNST growth and sensitized MPNSTs to standard-of-care ST:STUDY_SUMMARY chemotherapy. Knockdown of the redox-regulated transcription factor NRF2 slowed ST:STUDY_SUMMARY MPNST growth and decreased G6PD transcription. Analysis of patient MPNSTs ST:STUDY_SUMMARY identified a NRF2 gene signature correlating with tumor transformation. ST:STUDY_SUMMARY Furthermore, G6PD and NRF2 expression in pan-cancer TCGA samples correlates with ST:STUDY_SUMMARY patient survival. This work identifies NRF2-PPP dependency as a targetable ST:STUDY_SUMMARY vulnerability in these difficult to treat MPNSTs, particularly in the ST:STUDY_SUMMARY NF1/CDKN2A-deleted majority. ST:INSTITUTE University of Iowa ST:LABORATORY Eric Taylor Lab ST:LAST_NAME McGivney ST:FIRST_NAME Gavin ST:ADDRESS 3316 Pappajohn Biomedical Discovery Building, Iowa City, IA, 52242, USA ST:EMAIL gavin-mcgivney@uiowa.edu ST:PHONE 5158646586 ST:SUBMIT_DATE 2025-05-06 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENOTYPE_STRAIN Wildtype 129 Sv/Jae mice SU:CELL_STRAIN_DETAILS primary Malignant peripheral nerve sheath tumor derived cell lines SU:CELL_PASSAGE_NUMBER 20 #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS - GC_Blank Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_Blank.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC1 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC1.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC2 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC2.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC3 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC3.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC4 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC4.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC5 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC5.mzXML SUBJECT_SAMPLE_FACTORS - GC_QC6 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=GC-MS; RAW_FILE_NAME(Raw file name)=GC_QC6.mzXML SUBJECT_SAMPLE_FACTORS - LC_Blank1 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_Blank1.mzXML SUBJECT_SAMPLE_FACTORS - LC_Blank2 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_Blank2.mzXML SUBJECT_SAMPLE_FACTORS - LC_Blank3 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_Blank3.mzXML SUBJECT_SAMPLE_FACTORS - LC_Blank4 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_Blank4.mzXML SUBJECT_SAMPLE_FACTORS - LC_QC1 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_QC1.mzXML SUBJECT_SAMPLE_FACTORS - LC_QC2 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_QC2.mzXML SUBJECT_SAMPLE_FACTORS - LC_QC3 Sample source:Primary mouse MPNST cell line | Genotype:- MS method=LC-MS; RAW_FILE_NAME(Raw file name)=LC_QC3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.1.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.1.4.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.2.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.2.4.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.Cdkn2a.3.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.Cdkn2a.3.4.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.1.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.1.4.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.2.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.2.4.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.1 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.1.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.1.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.1.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.2 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.2.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.2.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.2.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.3 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.3.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.3.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.3.GC.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.4 Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=LC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.4.mzXML SUBJECT_SAMPLE_FACTORS - Nf1.p53.3.4.GC Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO MS method=GC-MS; RAW_FILE_NAME(Raw file name)=Nf1.p53.3.4.GC.mzXML #COLLECTION CO:COLLECTION_SUMMARY To engineer CRISPR/Cas9-driven tumor models containing CMV promoters for Cas9, CO:COLLECTION_SUMMARY sgRNAs targeting NF1, p16, and p19 or NF1 and p53 were assembled together with CO:COLLECTION_SUMMARY individual U6 promoters and sgRNA scaffold sequences using the Multiplex CO:COLLECTION_SUMMARY CRISPR/Cas9 Assembly System (Addgene Kit #1000000055). p16 (Ink4a) and p19 (ARF) CO:COLLECTION_SUMMARY are the 2 gene products of the gene, CDKN2A. P16 and p19 have alternative first CO:COLLECTION_SUMMARY exons, creating two unique RNA and protein products. Both p16 and p19 have CO:COLLECTION_SUMMARY canonical cell cycle roles, however, they have been attributed to non-cell cycle CO:COLLECTION_SUMMARY roles, including metabolism. After assembly into the A1x3 vector, the three CO:COLLECTION_SUMMARY sgRNAs were cloned into the Ad5CMVspCas9 shuttle plasmid (University of Iowa CO:COLLECTION_SUMMARY Viral Vector Core #G1272) as a single piece of DNA using traditional restriction CO:COLLECTION_SUMMARY enzyme cloning (ClaI and MluI sites). Adenoviruses were produced at the CO:COLLECTION_SUMMARY University of Iowa Viral Vector core as previously reported. Prior to injection, CO:COLLECTION_SUMMARY virus was mixed with DMEM and calcium phosphate as previously described. To CO:COLLECTION_SUMMARY induce tumorigenesis, 25µL of prepared virus was injected into the left flank CO:COLLECTION_SUMMARY sciatic nerve of wildtype 129 Sv/Jae mice. Tumor initiation was declared at a CO:COLLECTION_SUMMARY measurable volume of ~150-200 mm3. Tumors were measured 3 times per week by CO:COLLECTION_SUMMARY caliper. Tumor volumes were calculated using the formula V = (π × L × W × CO:COLLECTION_SUMMARY H)/6, with L, W, and H representing the length, width, and height of the tumor CO:COLLECTION_SUMMARY in mm, respectively. Tumors were harvested at a terminal size of 1500 mm3 or CO:COLLECTION_SUMMARY earlier if animals showed signs of distress in accordance with University of CO:COLLECTION_SUMMARY Iowa IACUC guidelines. Tissue was collected for histology, RNA, and generation CO:COLLECTION_SUMMARY of cell lines. tumor tissue was rinsed with 1X PBS and finely minced with CO:COLLECTION_SUMMARY dissection scissors in a 6-well plate. 5mL of Dissociation Buffer [Collagenase CO:COLLECTION_SUMMARY Type IV (700 units/mL, Gibco, 17104-019) and dispase (65 mg/mL, Gibco, CO:COLLECTION_SUMMARY 17105-041) was added to each well. The plate was incubated at 37 °C for one CO:COLLECTION_SUMMARY hour on orbital shaker and transferred to tissue culture hood. Tissue slurry was CO:COLLECTION_SUMMARY pipetted through 70 µM cell strainer (Fisherbrand) and collected in 50 mL CO:COLLECTION_SUMMARY conical tube. Cell strainer was washed with 25 mL of 1X PBS and collected in the CO:COLLECTION_SUMMARY same 50 mL conical tube. During this wash, the plunger from a 1 mL syringe (BD, CO:COLLECTION_SUMMARY 309628), was used to aid in collecting as much material as possible. The CO:COLLECTION_SUMMARY filtered suspension was centrifuged and supernatant decanted. Cell pellets were CO:COLLECTION_SUMMARY resuspended in DMEM [(Gibco, 11965-092), + 10% FBS (R&D Systems, S11150), 1% CO:COLLECTION_SUMMARY penicillin–streptomycin (Gibco, 15140-122) and 1% sodium pyruvate (Gibco, CO:COLLECTION_SUMMARY 11360-070)] and plated in 2 wells of a 6-well plate. Once 80-90% confluent, 33% CO:COLLECTION_SUMMARY of cells were transferred to a 10 cm dish. Cells were passaged at least 20 times CO:COLLECTION_SUMMARY before experimental analyses to ensure pure tumor populations. Cell culture CO:COLLECTION_SUMMARY plates were washed twice with ice-cold PBS followed by two washes with ice-cold CO:COLLECTION_SUMMARY water before being flash frozen using liquid nitrogen. Cell culture plates were CO:COLLECTION_SUMMARY lyophilized overnight. CO:SAMPLE_TYPE primary MPNST mouse cells #TREATMENT TR:TREATMENT_SUMMARY No treatment. This is a genotype study. Cells were maintained in DMEM [(Gibco, TR:TREATMENT_SUMMARY 11965-092), + 10% FBS (R&D Systems, S11150), 1% penicillin–streptomycin TR:TREATMENT_SUMMARY (Gibco, 15140-122) and 1% sodium pyruvate (Gibco, 11360-070)] #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lyophilized cells were scraped into 1 ml of ice-cold 2:2:1 SP:SAMPLEPREP_SUMMARY methanol/acetonitrile/water containing a mixture of internal standards SP:SAMPLEPREP_SUMMARY (D4-citric acid, D4-succinic acid, D8-valine, and U13C-labeled glutamine, SP:SAMPLEPREP_SUMMARY glutamic acid, lysine, methionine, serine, and tryptophan; Cambridge Isotope SP:SAMPLEPREP_SUMMARY Laboratories) to extract metabolites. The mixtures were transferred to a SP:SAMPLEPREP_SUMMARY microcentrifuge tube and flash frozen in liquid nitrogen, thawed for 10 minutes SP:SAMPLEPREP_SUMMARY in a water bath sonicator, and rotated for 1 hour at –20°C. Mixtures were SP:SAMPLEPREP_SUMMARY centrifuged for 10 minutes at 21,000 x g and 300 µl (GC-MS) or 200 µl (LC-MS) SP:SAMPLEPREP_SUMMARY of the cleared metabolite extracts were transferred to autosampler vials (GC-MS) SP:SAMPLEPREP_SUMMARY or microcentrifuge tubes (LC-MS) and dried using a SpeedVac vacuum concentrator SP:SAMPLEPREP_SUMMARY (Thermo). For GC-MS analysis, dried metabolite extracts were reconstituted in 20 SP:SAMPLEPREP_SUMMARY μl of 11.4 mg/ml methoxyamine (MOX) in anhydrous pyridine, vortexed for 5 SP:SAMPLEPREP_SUMMARY minutes, and heated for 1 hour at 60°C. Next, 16 μl of SP:SAMPLEPREP_SUMMARY N,O-Bis(trimethylsilyl)trifluoroacetamide (TMS) was added to each sample, which SP:SAMPLEPREP_SUMMARY were vortexed for 1 minute, and heated for 30 minutes at 60°C (47, 48). For SP:SAMPLEPREP_SUMMARY LC-MS, dried extracts were reconstituted in 20 μl of acetonitrile/water (1:1 SP:SAMPLEPREP_SUMMARY v/v) vortexed well, rotated on a rotator in −20 °C overnight, centrifuged and SP:SAMPLEPREP_SUMMARY the supernatant was transferred to LC-MS autosampler vials for analysis. LC-MS SP:SAMPLEPREP_SUMMARY data were acquired on a Thermo Q Exactive hybrid quadrupole Orbitrap mass SP:SAMPLEPREP_SUMMARY spectrometer with a Vanquish Flex UHPLC system or Vanquish Horizon UHPLC system. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY The LC column used was a Millipore SeQuant ZIC-pHILIC (2.1 mm by 150 mm, 5-μm CH:CHROMATOGRAPHY_SUMMARY particle size) with a ZIC-pHILIC guard column (20 mm by 2.1 mm). The injection CH:CHROMATOGRAPHY_SUMMARY volume was 2 to 5 μl. The mobile phase comprised buffer A [20 mM (NH4)2CO3, CH:CHROMATOGRAPHY_SUMMARY 0.1% NH4OH (v/v)] and buffer B [acetonitrile]. The chromatographic gradient was CH:CHROMATOGRAPHY_SUMMARY run at a flow rate of 0.150 ml/min as follows: 0-to 21-min linear gradient from CH:CHROMATOGRAPHY_SUMMARY 80 to 20% buffer B; 20-to 20.5-min linear gradient from 20 to 80% buffer B; and CH:CHROMATOGRAPHY_SUMMARY 20.5-to 28-min hold at 80% buffer B. The CH:METHODS_FILENAME GC_LC-MS_analysis_protocol.txt CH:INSTRUMENT_NAME Thermo Vanquish Flex UHPLC CH:COLUMN_NAME Millipore SeQuant ZIC-pHILIC (150 x 2.1mm, 5um) with a ZIC-pHILIC (20 x 2.1mm) CH:COLUMN_NAME guard column CH:COLUMN_TEMPERATURE Room temperature (~23 degrees C) CH:FLOW_GRADIENT 0-to 21min linear gradient from 80 to 20% buffer B; 20-to 20.5min linear CH:FLOW_GRADIENT gradient from 20 to 80% buffer B; and 20.5-to 28min hold at 80% buffer B CH:FLOW_RATE 0.150 mL/min CH:SOLVENT_A 100% water; 20 mM ammonium carbonate; 0.1% ammonium hydroxide CH:SOLVENT_B 100% acetonitrile CH:CHROMATOGRAPHY_TYPE HILIC #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE GC_LC-MS_analysis_protocol.txt #MS MS:INSTRUMENT_NAME Thermo Q Exactive hybrid quadrupole Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS MS operated in positive or negative polarity switching fullscan mode with the MS:MS_COMMENTS spray voltage set to 3.0 kV, the heated capillary held at 275°C, and the Heated MS:MS_COMMENTS Electrospray Ionization (HESI) probe held at 350°C. The sheath gas flow was set MS:MS_COMMENTS to 40 units, the auxiliary gas flow was set to 15 units, and the sweep gas flow MS:MS_COMMENTS was set to 1 unit. MS data resolution was set at 70,000, the Automatic Gain MS:MS_COMMENTS Control (AGC) target at 1e6, and a maximum injection time of 200 ms. MS:ION_MODE UNSPECIFIED #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Intensity MS_METABOLITE_DATA_START Samples LC_QC1 LC_QC2 LC_QC3 Nf1.Cdkn2a.1.1 Nf1.Cdkn2a.1.2 Nf1.Cdkn2a.1.3 Nf1.Cdkn2a.1.4 Nf1.Cdkn2a.2.1 Nf1.Cdkn2a.2.2 Nf1.Cdkn2a.2.3 Nf1.Cdkn2a.2.4 Nf1.Cdkn2a.3.1 Nf1.Cdkn2a.3.2 Nf1.Cdkn2a.3.3 Nf1.Cdkn2a.3.4 Nf1.p53.1.1 Nf1.p53.1.2 Nf1.p53.1.3 Nf1.p53.1.4 Nf1.p53.2.1 Nf1.p53.2.2 Nf1.p53.2.3 Nf1.p53.2.4 Nf1.p53.3.1 Nf1.p53.3.2 Nf1.p53.3.3 Nf1.p53.3.4 Factors Sample source:Primary mouse MPNST cell line | Genotype:- Sample source:Primary mouse MPNST cell line | Genotype:- Sample source:Primary mouse MPNST cell line | Genotype:- Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_Cdkn2a_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO Sample source:Primary mouse MPNST cell line | Genotype:Nf1_p53_dKO 2-Deoxyadenosine 5-monophosphate 1129673.0000 1195630.0000 1323202.0000 628760.0000 572313.0000 424853.0000 733794.0000 577807.0000 654911.0000 522142.0000 713247.0000 227783.0000 216447.0000 257604.0000 335068.0000 1326699.0000 1551939.0000 1567554.0000 1900814.0000 1986925.0000 1378736.0000 1580443.0000 1564608.0000 2582825.0000 3847573.0000 2100163.0000 2043698.0000 2-Deoxyguanosine 5-triphosphate sodium salt hydrate 279920015.0000 349007919.0000 460190772.0000 127176706.0000 171117488.0000 109533054.0000 178311095.0000 321727241.0000 352782013.0000 327619736.0000 396998925.0000 480708843.0000 346153651.0000 354026915.0000 395732100.0000 1051463332.0000 482329348.0000 855086095.0000 805264307.0000 416609386.0000 402744140.0000 363297210.0000 329103027.0000 87967646.0000 166516947.0000 95866608.0000 115478727.0000 3-Hydroxy-3-methylglutaric acid 3652952.0000 3485933.0000 3655685.0000 1814009.0000 2090150.0000 2033445.0000 111555.0000 9066478.0000 7591732.0000 8102170.0000 12248454.0000 3480913.0000 3481369.0000 4155279.0000 3016229.0000 6253313.0000 5387126.0000 7128708.0000 8210750.0000 3862063.0000 3024086.0000 3091031.0000 4636628.0000 3280375.0000 3221947.0000 3364987.0000 2971323.0000 4-Imidazoleacrylic acid 675394534.0000 706222262.0000 727341056.0000 297749468.0000 818175213.0000 343947852.0000 293313051.0000 218867021.0000 240000484.0000 375472313.0000 344800825.0000 314535725.0000 269490046.0000 381939986.0000 381054159.0000 1732746470.0000 530817430.0000 592460518.0000 113552827.0000 384161005.0000 209677516.0000 670487482.0000 666978770.0000 267909331.0000 372306038.0000 202523918.0000 246356521.0000 5-Deoxy-5-(methylthio)adenosine 340799522.0000 438206186.0000 436377113.0000 183688745.0000 189557960.0000 170538096.0000 221860126.0000 398199027.0000 300359821.0000 341953127.0000 462123479.0000 341573255.0000 319755014.0000 313957844.0000 262390200.0000 859765244.0000 907544272.0000 943835706.0000 912831292.0000 435556477.0000 416460487.0000 431534401.0000 394649040.0000 402245113.0000 497194069.0000 396977979.0000 368711346.0000 6-phosphogluconate 3344282.0000 4226775.0000 4896147.0000 2319377.0000 2569509.0000 2234769.0000 2735675.0000 7197252.0000 7729843.0000 6086862.0000 8237276.0000 2825295.0000 2613524.0000 2514100.0000 2556119.0000 8601194.0000 6586477.0000 8492162.0000 7727222.0000 2590985.0000 2110859.0000 2665763.0000 1976753.0000 1047313.0000 1668885.0000 1350435.0000 1320069.0000 ADP 35312423.0000 38293711.0000 45017017.0000 9636491.0000 12676883.0000 8557208.0000 22863395.0000 27995857.0000 24550432.0000 28307153.0000 33521013.0000 42742570.0000 33080674.0000 35881226.0000 41594049.0000 99293267.0000 54842812.0000 83249431.0000 75039426.0000 45676400.0000 44607330.0000 41283026.0000 37437669.0000 11097552.0000 11130104.0000 8037007.0000 10244435.0000 Allantoin 2724845.0000 4196739.0000 4109179.0000 2404721.0000 2746665.0000 2008029.0000 4079052.0000 2145817.0000 2305503.0000 2890356.0000 3285760.0000 2719183.0000 2947461.0000 3636380.0000 2938806.0000 4934520.0000 5434197.0000 4766200.0000 4953150.0000 6425996.0000 5060755.0000 6052353.0000 4819669.0000 3033072.0000 5049222.0000 3762458.0000 3904234.0000 AMP 116386482.0000 137705437.0000 141674152.0000 107096241.0000 97773738.0000 92270548.0000 139720822.0000 47484923.0000 43727402.0000 33610205.0000 48985671.0000 41891218.0000 35707227.0000 37024593.0000 42618850.0000 91695780.0000 92703534.0000 111986100.0000 128615321.0000 249914992.0000 156800013.0000 216680858.0000 212440117.0000 405489324.0000 475930867.0000 300262409.0000 301528237.0000 ATP 279920015.0000 349007919.0000 460190772.0000 127176706.0000 171117488.0000 109533054.0000 178311095.0000 321727241.0000 352782013.0000 327619736.0000 396998925.0000 480708843.0000 346153651.0000 354026915.0000 395732100.0000 1051463332.0000 482329348.0000 855086095.0000 805264307.0000 416609386.0000 402744140.0000 363297210.0000 329103027.0000 87967646.0000 166516947.0000 95866608.0000 115478727.0000 Betaine 939573728.0000 989870498.0000 1038261955.0000 357099818.0000 409514461.0000 434287322.0000 606751262.0000 651235052.0000 784380061.0000 658110702.0000 829104691.0000 449813511.0000 444524808.0000 592153786.0000 593805030.0000 1737772850.0000 2136484375.0000 2035342359.0000 2018268875.0000 1427239487.0000 885226445.0000 1280252691.0000 964509050.0000 583710834.0000 855482810.0000 839605346.0000 939485021.0000 B-Nicotinamide mononucleotide 1052536.0000 821285.0000 738121.0000 821008.0000 826285.0000 568587.0000 905672.0000 1083532.0000 1293944.0000 1320914.0000 1929875.0000 781171.0000 795390.0000 839576.0000 747520.0000 622164.0000 313325.0000 521538.0000 770008.0000 1080492.0000 1325112.0000 1261091.0000 692319.0000 840351.0000 320625.0000 544567.0000 627683.0000 Butyryl-L-carnitine 172845327.0000 177471065.0000 188977862.0000 45605041.0000 44804309.0000 47916884.0000 62021536.0000 136619961.0000 143789218.0000 129676811.0000 160361522.0000 78055457.0000 82587387.0000 83149543.0000 90036177.0000 334608834.0000 384952205.0000 342708715.0000 333429435.0000 145192672.0000 123621783.0000 133152766.0000 129355462.0000 73633886.0000 94678618.0000 74895540.0000 76965451.0000 Citrate 303475044.0000 354687745.0000 386848061.0000 150947411.0000 166813765.0000 114701773.0000 196815521.0000 365840574.0000 386880872.0000 302340021.0000 412050648.0000 306298706.0000 320857874.0000 265104940.0000 310346220.0000 513468031.0000 570124230.0000 545290774.0000 563984911.0000 323843147.0000 311875718.0000 272446031.0000 305327203.0000 217460588.0000 300576570.0000 184519337.0000 195492659.0000 Citrulline 61167438.0000 63471298.0000 63897059.0000 27551329.0000 31507821.0000 39044178.0000 33500370.0000 28320048.0000 29883559.0000 33223151.0000 35454022.0000 19916313.0000 26033783.0000 29977274.0000 37220788.0000 101360446.0000 91488427.0000 77699737.0000 78939755.0000 56522580.0000 34833876.0000 76190757.0000 50639496.0000 31431611.0000 40367947.0000 39318646.0000 44118838.0000 CMP 9007220.0000 9574846.0000 9550908.0000 5319311.0000 5199615.0000 4833905.0000 6431813.0000 4521140.0000 4214174.0000 3095201.0000 3063396.0000 4431133.0000 4047499.0000 4359601.0000 4599485.0000 5188799.0000 3852038.0000 4619488.0000 5123536.0000 18256492.0000 12787981.0000 16252613.0000 16127432.0000 31131510.0000 31181053.0000 22759216.0000 21154904.0000 Creatine 17235847536.0000 17808953116.0000 18342349371.0000 7543975428.0000 7874785787.0000 8147372502.0000 9811041722.0000 9975012572.0000 10842511488.0000 9889260642.0000 11219772225.0000 15820454979.0000 15251545885.0000 17099127009.0000 17653684267.0000 20803032634.0000 20389678834.0000 20856022605.0000 20821988688.0000 24782987185.0000 20819558987.0000 23370665638.0000 21345768013.0000 15903722968.0000 20646400458.0000 17780528755.0000 16583040731.0000 Cytidine 989942.0000 1145544.0000 1337121.0000 280856.0000 250815.0000 250684.0000 335807.0000 365553.0000 569803.0000 320155.0000 561074.0000 290284.0000 372284.0000 329915.0000 460894.0000 621386.0000 664732.0000 411414.0000 358043.0000 1104704.0000 748397.0000 801691.0000 918838.0000 5020551.0000 4199717.0000 3536925.0000 3209530.0000 Cytidine 5-diphosphocholine sodium salt dihydrate 12791500.0000 14566135.0000 14148230.0000 5943583.0000 6764644.0000 4893176.0000 7735415.0000 8312321.0000 8835714.0000 6031974.0000 7765889.0000 6794575.0000 7147416.0000 6844314.0000 7758790.0000 10047838.0000 10485708.0000 9960283.0000 9709051.0000 37430565.0000 24653165.0000 34307011.0000 29843990.0000 23835883.0000 24774083.0000 18435294.0000 15664408.0000 Decanoyl-L-carnitine 563297.0000 665029.0000 776089.0000 569479.0000 953762.0000 1086136.0000 476178.0000 70947.0000 673897.0000 164175.0000 769492.0000 653565.0000 615226.0000 516072.0000 885494.0000 738217.0000 997223.0000 554538.0000 186991.0000 1028009.0000 889593.0000 497351.0000 1179300.0000 166777.0000 375379.0000 546384.0000 1251315.0000 D-Gluconic acid solution 428101348.0000 520560837.0000 543876467.0000 161912435.0000 171861961.0000 157866940.0000 262252300.0000 262938909.0000 261749684.0000 264781638.0000 363065216.0000 325392017.0000 292532917.0000 343226624.0000 397389311.0000 1017066704.0000 1023449787.0000 1012118358.0000 1052516629.0000 914512780.0000 562680140.0000 843923816.0000 658647808.0000 458096294.0000 504957893.0000 469300044.0000 475884391.0000 D-Glucosamine 6-phosphate sodium salt 4658284.0000 4744238.0000 4959894.0000 2598544.0000 2732429.0000 2470878.0000 3047931.0000 4598680.0000 4496272.0000 4602666.0000 5117751.0000 3249503.0000 3350664.0000 3348793.0000 3551894.0000 8200859.0000 8969309.0000 8881211.0000 9135071.0000 6078222.0000 5469179.0000 5883367.0000 5520009.0000 3691636.0000 3305664.0000 3622536.0000 3425815.0000 D-Glucose 6-phosphate potassium salt 47962921.0000 54348477.0000 57390443.0000 40946371.0000 41740226.0000 36705673.0000 48619763.0000 82626805.0000 75858384.0000 57682432.0000 75301804.0000 51459031.0000 45946649.0000 52442757.0000 52165505.0000 68203325.0000 64420697.0000 75408071.0000 79330851.0000 32716135.0000 27993155.0000 30016664.0000 27384640.0000 36337415.0000 59056118.0000 44470636.0000 43209499.0000 DL-2-Aminoadipic acid 11590679.0000 12547591.0000 12054435.0000 7503374.0000 7506244.0000 7685646.0000 9654615.0000 7379911.0000 8640776.0000 6565227.0000 7796489.0000 10036638.0000 9177920.0000 11729162.0000 12824127.0000 19755936.0000 12250395.0000 10986044.0000 11003200.0000 12349767.0000 8893360.0000 12370484.0000 11068247.0000 17655717.0000 15051824.0000 19087328.0000 20874428.0000 DL-lauroylcarnitine 855352.0000 1701736.0000 514604.0000 623733.0000 725259.0000 429304.0000 398502.0000 190172.0000 320412.0000 233521.0000 279276.0000 393151.0000 1028112.0000 1064872.0000 737798.0000 509388.0000 1145684.0000 1467174.0000 377755.0000 1748843.0000 1674493.0000 2655051.0000 1722523.0000 957131.0000 804504.0000 990012.0000 1679123.0000 D-(+)-Trehalose dihydrate 7776874.0000 8144373.0000 7685805.0000 4649071.0000 13107804.0000 2774695.0000 4094318.0000 3412002.0000 5435040.0000 4216758.0000 4969640.0000 6536458.0000 4205402.0000 5735720.0000 4957828.0000 10171038.0000 4557164.0000 3747403.0000 702337.0000 2745414.0000 2554569.0000 7522009.0000 8421918.0000 2887828.0000 6807138.0000 2244313.0000 2578643.0000 FAD 3509710.0000 4411349.0000 4631709.0000 2904772.0000 2975448.0000 2625563.0000 3297275.0000 3633669.0000 3361203.0000 2577725.0000 3966195.0000 3444403.0000 3088390.0000 2585091.0000 3249859.0000 7503448.0000 5563911.0000 9050558.0000 8570213.0000 4585434.0000 3712139.0000 4416355.0000 4012526.0000 3528478.0000 3756939.0000 3483241.0000 3475433.0000 GDP 12549730.0000 14814850.0000 15462810.0000 8988973.0000 10438757.0000 8416016.0000 10874002.0000 12076280.0000 12136377.0000 9157045.0000 12527564.0000 11535565.0000 9971364.0000 9282846.0000 11254843.0000 14993139.0000 9516168.0000 17854675.0000 18581896.0000 24564912.0000 16681381.0000 21968535.0000 18874491.0000 20977653.0000 20863698.0000 17269354.0000 19851087.0000 Glutathione (oxidized) 52824312.0000 64042416.0000 74609353.0000 24910278.0000 19890221.0000 10475038.0000 88450415.0000 24025879.0000 32480230.0000 22855118.0000 40003920.0000 31917035.0000 29738385.0000 47610652.0000 49924335.0000 90596224.0000 83639268.0000 134990157.0000 128474095.0000 102950348.0000 87104461.0000 85251140.0000 52977415.0000 67900248.0000 81730126.0000 56397718.0000 99134247.0000 Glutathione (reduced) 1452083757.0000 1581559207.0000 1615405395.0000 814203436.0000 926610925.0000 854190972.0000 873661146.0000 1379218915.0000 1476429645.0000 1206659362.0000 1391211013.0000 1229867780.0000 1179189818.0000 1189247510.0000 1315547475.0000 2172882451.0000 2532893150.0000 2367201912.0000 2297061093.0000 1615855402.0000 1483150890.0000 1703307067.0000 1606040274.0000 1788542296.0000 2064911249.0000 2032479445.0000 1865523022.0000 GMP 6749056.0000 7616433.0000 7966520.0000 4564115.0000 4356764.0000 4747614.0000 6170553.0000 2624608.0000 2248685.0000 1689068.0000 2455623.0000 2909579.0000 2496049.0000 2734460.0000 3155949.0000 4849191.0000 4156635.0000 5645257.0000 5893751.0000 16090991.0000 9215171.0000 13691941.0000 12207630.0000 28059697.0000 22808485.0000 20255418.0000 21570564.0000 GTP 15405624.0000 18041822.0000 25284058.0000 5376015.0000 7207726.0000 4849143.0000 7529417.0000 20856036.0000 25865804.0000 20711168.0000 28511514.0000 25228159.0000 18515463.0000 16339633.0000 21570872.0000 62732986.0000 23020729.0000 49061504.0000 46066038.0000 28545970.0000 24450249.0000 23475087.0000 21751345.0000 9898331.0000 14027037.0000 8302621.0000 9592518.0000 Guanosine 910470.0000 1041158.0000 1073638.0000 272756.0000 594924.0000 291493.0000 322771.0000 439150.0000 517328.0000 460767.0000 614650.0000 265175.0000 278403.0000 290502.0000 257251.0000 522066.0000 463718.0000 516106.0000 454815.0000 2772671.0000 1113861.0000 2198363.0000 2101497.0000 3888441.0000 2504088.0000 2552661.0000 2363596.0000 Hexanoyl-L-carnitine 3432518.0000 3589917.0000 3608195.0000 1408978.0000 1548809.0000 1353487.0000 2120685.0000 1975348.0000 1913776.0000 1936405.0000 2565258.0000 2051366.0000 2185358.0000 2377975.0000 2582558.0000 6555513.0000 7072701.0000 6213642.0000 6163356.0000 4597165.0000 3145106.0000 4522258.0000 4741578.0000 1977945.0000 1855079.0000 2167310.0000 2277824.0000 Hypotaurine 100323998.0000 99500607.0000 106241785.0000 71552688.0000 80768517.0000 88882087.0000 124042025.0000 69671930.0000 77007008.0000 71098046.0000 85851181.0000 116017175.0000 117739871.0000 148499227.0000 179898837.0000 103257802.0000 125894394.0000 115569410.0000 106334229.0000 118399732.0000 86604210.0000 110944107.0000 91217576.0000 86534911.0000 112025290.0000 102144861.0000 97879343.0000 IMP 15195748.0000 16146758.0000 16708565.0000 2683878.0000 4526165.0000 2833031.0000 1973008.0000 3936239.0000 3475977.0000 2739193.0000 3950625.0000 3172215.0000 3415070.0000 2820687.0000 2806468.0000 3309979.0000 2913635.0000 2970904.0000 3278763.0000 58099449.0000 26486346.0000 56744454.0000 44928333.0000 68008721.0000 43403176.0000 50380222.0000 47853806.0000 Inosine 7075742.0000 8819796.0000 9071454.0000 1788970.0000 6018886.0000 1402772.0000 1716977.0000 1665263.0000 1336479.0000 1096657.0000 1857026.0000 1773141.0000 1901837.0000 1919475.0000 1805899.0000 1418940.0000 1356499.0000 1540850.0000 1414242.0000 30573049.0000 11411299.0000 27244856.0000 22851160.0000 30232734.0000 18212628.0000 26117374.0000 23725256.0000 Isopentenyl pyrophosphate triammonium salt solution 112524.0000 215437.0000 218870.0000 92530.0000 90868.0000 37105.0000 56508.0000 220767.0000 243407.0000 238507.0000 254154.0000 157720.0000 102792.0000 121957.0000 151221.0000 705423.0000 595665.0000 800326.0000 790284.0000 27793.0000 32253.0000 22864.0000 53938.0000 4688.0000 20367.0000 8968.0000 4060.0000 L-Acetylcarnitine 439428538.0000 459033636.0000 481424595.0000 200468111.0000 210260214.0000 226058683.0000 311793996.0000 284771103.0000 279755281.0000 252496906.0000 318372549.0000 266946347.0000 275438369.0000 292581178.0000 311362473.0000 615767451.0000 719687724.0000 736109579.0000 791856901.0000 495082479.0000 289117366.0000 429227854.0000 355990632.0000 458623610.0000 341603685.0000 405208669.0000 420799449.0000 L-Carnitine 1168810911.0000 1162438367.0000 1218901309.0000 456391731.0000 499359009.0000 522127676.0000 715670486.0000 428068919.0000 450594122.0000 396629380.0000 510885361.0000 576637375.0000 562751122.0000 677037238.0000 763901871.0000 1699488041.0000 1735778818.0000 1561838778.0000 1562466351.0000 1615596819.0000 1245490816.0000 1549806084.0000 1271104964.0000 759417907.0000 1023901416.0000 970110260.0000 978757413.0000 L-Carnosine 14473245.0000 13361134.0000 13789506.0000 9978249.0000 11024829.0000 16281238.0000 14480899.0000 5170201.0000 6634698.0000 7188619.0000 8234761.0000 6972868.0000 8272342.0000 8990236.0000 9203869.0000 11979716.0000 15476844.0000 11109601.0000 14352113.0000 11708858.0000 10088137.0000 12273979.0000 9822869.0000 12101809.0000 12956528.0000 12755026.0000 12437850.0000 L-Cystathionine 1827718.0000 2148457.0000 1860697.0000 1495994.0000 1564018.0000 1922577.0000 2546926.0000 1192683.0000 1307634.0000 1206513.0000 1245259.0000 3101427.0000 3743675.0000 4671766.0000 4778830.0000 727097.0000 1513468.0000 1209519.0000 933977.0000 1360375.0000 774051.0000 1253952.0000 884869.0000 637288.0000 761386.0000 1166848.0000 1345386.0000 L-Dihydroorotic acid 845938.0000 1458489.0000 1400804.0000 1041486.0000 962730.0000 959329.0000 980356.0000 2196719.0000 1223644.0000 1387945.0000 1428980.0000 1410787.0000 1131055.0000 1137090.0000 913023.0000 1465601.0000 1060532.0000 1972636.0000 1761755.0000 1129271.0000 1171248.0000 1247373.0000 1446958.0000 1404543.0000 2336678.0000 1572704.0000 1472861.0000 L-Homoserine 624815905.0000 653119660.0000 670042332.0000 344324427.0000 367116861.0000 408085053.0000 538348061.0000 582358738.0000 627971240.0000 626861350.0000 695483264.0000 377555176.0000 355383679.0000 462152956.0000 489037242.0000 977043705.0000 985487650.0000 928958475.0000 911140123.0000 940597397.0000 691488697.0000 868133319.0000 695545794.0000 518227343.0000 654066983.0000 719708578.0000 765693986.0000 L-Kynurenine 2045605.0000 2363673.0000 2399987.0000 1044902.0000 1082671.0000 1134441.0000 1968697.0000 1601631.0000 1981324.0000 1947083.0000 2396252.0000 1150784.0000 1220947.0000 1613557.0000 1526593.0000 4358444.0000 4542455.0000 4670030.0000 4899232.0000 3739206.0000 2706639.0000 3294884.0000 2187678.0000 1298557.0000 2087597.0000 1896841.0000 2062755.0000 L-(-)-Malic acid 662303005.0000 647578851.0000 636190780.0000 447093053.0000 437053503.0000 441797160.0000 527629402.0000 986272851.0000 1027698355.0000 881007271.0000 1043133894.0000 715408277.0000 730694363.0000 746823058.0000 767202623.0000 591584360.0000 614769953.0000 589956070.0000 573543756.0000 619321744.0000 603038398.0000 679139683.0000 640941693.0000 550340831.0000 586750451.0000 550605188.0000 482483768.0000 L-Norvaline 939573728.0000 989870498.0000 1038261955.0000 357099818.0000 409514461.0000 434287322.0000 606751262.0000 651235052.0000 784380061.0000 658110702.0000 829104691.0000 449813511.0000 444524808.0000 592153786.0000 593805030.0000 1737772850.0000 2136484375.0000 2035342359.0000 2018268875.0000 1427239487.0000 885226445.0000 1280252691.0000 964509050.0000 583710834.0000 855482810.0000 839605346.0000 939485021.0000 L-(+)-Tartaric acid 6081725.0000 7281933.0000 8589032.0000 11977939.0000 11075402.0000 9627412.0000 7831573.0000 8062803.0000 8027763.0000 2676907.0000 5048471.0000 7293722.0000 7445914.0000 10492226.0000 10000771.0000 6421254.0000 5225364.0000 11603051.0000 11297401.0000 7408445.0000 7929664.0000 9569084.0000 7185460.0000 7583519.0000 9897415.0000 7085300.0000 5232455.0000 Malonic acid 5066912.0000 4760956.0000 5142568.0000 2209534.0000 2884883.0000 2088737.0000 2063980.0000 2984045.0000 2129576.0000 2125821.0000 1924617.0000 3523883.0000 3358152.0000 4796414.0000 4098806.0000 3564247.0000 2566341.0000 3345348.0000 2746276.0000 5098744.0000 3955584.0000 6108986.0000 4741484.0000 3846353.0000 4703567.0000 3229176.0000 3792906.0000 Myristoyl-L-carnitine 13163577.0000 13363642.0000 14058082.0000 9001691.0000 9073877.0000 10288636.0000 11416935.0000 2814274.0000 4154160.0000 5321520.0000 4265007.0000 12206158.0000 11714095.0000 12832025.0000 15157218.0000 7779275.0000 7194946.0000 7577622.0000 6305483.0000 26497645.0000 15949632.0000 26572270.0000 44040039.0000 7751790.0000 7510352.0000 9834502.0000 10321248.0000 N-acetyl-neuraminic acid 12914290.0000 16270387.0000 17390443.0000 5087890.0000 5307088.0000 5452085.0000 8562292.0000 10620856.0000 11398365.0000 10689135.0000 14435657.0000 6046992.0000 5332528.0000 6487141.0000 6914172.0000 33476694.0000 33554467.0000 32971024.0000 36000779.0000 12876290.0000 7917581.0000 11629198.0000 8694841.0000 6366682.0000 9101313.0000 7354326.0000 7594206.0000 NAD 213229741.0000 230286012.0000 232437662.0000 129452288.0000 138724035.0000 111847807.0000 149655236.0000 227519359.0000 246301159.0000 210931766.0000 273516556.0000 170718286.0000 169130081.0000 166089602.0000 169070859.0000 334305148.0000 311106837.0000 326599569.0000 322979903.0000 217703026.0000 205620502.0000 215496240.0000 215091000.0000 163893054.0000 195880670.0000 167830862.0000 161841264.0000 NADH 11254152.0000 11836635.0000 11238236.0000 3348486.0000 3940063.0000 2758340.0000 4415125.0000 11829232.0000 11915707.0000 11410843.0000 16084922.0000 10175813.0000 9482836.0000 8553021.0000 9176283.0000 20684028.0000 25058615.0000 20282733.0000 21663714.0000 11810393.0000 9543479.0000 12881188.0000 12859269.0000 8199723.0000 9734575.0000 7738041.0000 5368192.0000 NADP 6238318.0000 8444358.0000 10655149.0000 2685971.0000 3007205.0000 1953731.0000 4602867.0000 4934982.0000 6315979.0000 5420612.0000 7578847.0000 4339756.0000 4405859.0000 5059006.0000 5479414.0000 15416933.0000 14019825.0000 17017955.0000 16233986.0000 8540012.0000 5314767.0000 6135387.0000 4314690.0000 4806587.0000 12385549.0000 5863185.0000 8695129.0000 NADPH 4772196.0000 4285855.0000 3316569.0000 3415623.0000 3427488.0000 2813233.0000 3152039.0000 8540912.0000 8686474.0000 5204036.0000 8416893.0000 9109800.0000 7938392.0000 6507107.0000 7275774.0000 2868553.0000 253482.0000 2821758.0000 1706527.0000 4858078.0000 6447519.0000 7244511.0000 7875730.0000 5188680.0000 3467473.0000 339833.0000 N(alpha)-Acetyl-L-asparagine 1715015.0000 2887451.0000 2944271.0000 1361830.0000 1263421.0000 852204.0000 5574556.0000 876598.0000 731996.0000 1041736.0000 1438728.0000 805788.0000 909152.0000 1569678.0000 1187559.0000 5163789.0000 2092349.0000 8476879.0000 6311861.0000 6514830.0000 5561772.0000 3791976.0000 2606691.0000 2409761.0000 2006925.0000 2434783.0000 5311860.0000 Nicotinamide 349494639.0000 355480855.0000 328975560.0000 173619310.0000 176358307.0000 170419265.0000 274423970.0000 196412443.0000 207992461.0000 229844795.0000 319612606.0000 184157410.0000 209824387.0000 241117916.0000 205883925.0000 492002687.0000 768381938.0000 494153244.0000 473140270.0000 414921995.0000 308759692.0000 416262146.0000 334857171.0000 254756318.0000 368180923.0000 295211641.0000 295921538.0000 Nicotinate 5024215.0000 4794814.0000 5316224.0000 3646762.0000 7942636.0000 3707936.0000 3926567.0000 3157345.0000 3200574.0000 3222571.0000 3992641.0000 3586616.0000 4054872.0000 3855460.0000 3795130.0000 5210673.0000 5163064.0000 5833586.0000 4656503.0000 5130785.0000 5408662.0000 6030534.0000 6389237.0000 2766005.0000 4134686.0000 3563352.0000 3495189.0000 N-Methyl-D-aspartic acid 1939807574.0000 2036048725.0000 2080112136.0000 1282169949.0000 1439251699.0000 1318196640.0000 1754760097.0000 1726562110.0000 1856241142.0000 1602968978.0000 1840516617.0000 2009027165.0000 1918358643.0000 2144695448.0000 2266926232.0000 2220325930.0000 2142593589.0000 2148219312.0000 2153455669.0000 2405932890.0000 2198549278.0000 2432443553.0000 2240127581.0000 1854279600.0000 2083830262.0000 2123053209.0000 2118390810.0000 Oleoyl-L-carnitine 23772670.0000 25549376.0000 25320497.0000 21666668.0000 21846014.0000 21137428.0000 27913188.0000 11893828.0000 11472344.0000 9855308.0000 13659817.0000 25824634.0000 26358022.0000 28046140.0000 33133219.0000 13525733.0000 13247384.0000 12878005.0000 10140404.0000 50764844.0000 27948504.0000 53337141.0000 39916503.0000 21214227.0000 17674970.0000 23039227.0000 23333826.0000 Palmitoyl-L-carnitine 37315330.0000 41021573.0000 40193152.0000 34461804.0000 30452080.0000 32066104.0000 37096775.0000 15559998.0000 15422169.0000 14552318.0000 18938593.0000 48103830.0000 49994759.0000 47559421.0000 51278632.0000 14974865.0000 23374260.0000 20251805.0000 18154853.0000 62528746.0000 44977208.0000 67421190.0000 55630066.0000 24637536.0000 26254998.0000 25994780.0000 23829477.0000 Phosphocreatine 1378857472.0000 1415288953.0000 1435568355.0000 703406257.0000 734998448.0000 711412274.0000 959274388.0000 619656364.0000 649134030.0000 555619341.0000 657880465.0000 1341773298.0000 1223118642.0000 1462586132.0000 1627144653.0000 1741896202.0000 1646745770.0000 1681988475.0000 1609120878.0000 2424910101.0000 2171112915.0000 2448422016.0000 2247531261.0000 744627487.0000 1076896610.0000 890954336.0000 857177109.0000 Propionyl-L-carnitine 144715728.0000 154484703.0000 157516545.0000 29495352.0000 36593647.0000 39776498.0000 57921062.0000 110408865.0000 104039742.0000 112742568.0000 132902494.0000 59937901.0000 75361759.0000 79949108.0000 86293644.0000 297414974.0000 241395023.0000 254091318.0000 258684531.0000 64423312.0000 48376661.0000 55050240.0000 49649153.0000 33490243.0000 36826589.0000 45425356.0000 48460296.0000 Pyridoxine 540561084.0000 563761178.0000 591587479.0000 251766808.0000 273630902.0000 240131149.0000 455263494.0000 388631218.0000 488452961.0000 475808500.0000 614740116.0000 337981514.0000 355361635.0000 450945255.0000 452315994.0000 877144121.0000 1190879901.0000 870728750.0000 875625667.0000 614808096.0000 406342801.0000 581411370.0000 408592308.0000 383861294.0000 603803573.0000 471519520.0000 492596977.0000 Ribose 5-phosphate 17794935.0000 21488139.0000 23405046.0000 5735922.0000 6911413.0000 5019635.0000 8904687.0000 16752450.0000 17154607.0000 13815682.0000 18447258.0000 11105469.0000 8130556.0000 10294304.0000 8887978.0000 43952609.0000 42325657.0000 43035037.0000 41198421.0000 23787066.0000 18588369.0000 22434118.0000 20978632.0000 20447175.0000 26156284.0000 19079684.0000 20109995.0000 S-(5-Adenosyl)-L-methionine p-toluenesulfonate salt 45365018.0000 47061986.0000 49491848.0000 36150499.0000 40457122.0000 37282289.0000 35945342.0000 60816106.0000 62472525.0000 56249336.0000 67804822.0000 42569654.0000 42051615.0000 36280140.0000 40662381.0000 60293332.0000 82357888.0000 72086983.0000 60561423.0000 30841504.0000 35976402.0000 35435904.0000 32983870.0000 60989371.0000 67611734.0000 60712943.0000 56489857.0000 Succinic acid 9188230.0000 12919385.0000 11262764.0000 5052867.0000 3590221.0000 2886377.0000 7214220.0000 11638837.0000 11862945.0000 10760380.0000 14315269.0000 5014317.0000 4372346.0000 6450556.0000 21140101.0000 24646992.0000 22902487.0000 25109446.0000 8896541.0000 11024335.0000 9298777.0000 6414462.0000 6418304.0000 8287829.0000 4460941.0000 4836044.0000 Taurine 1195817583.0000 1272828758.0000 1341498955.0000 1034772160.0000 1094600773.0000 1178282338.0000 1601740824.0000 699756179.0000 771667934.0000 820355960.0000 912979034.0000 1243924376.0000 1600332314.0000 1666508700.0000 1261534983.0000 1351388565.0000 1278797761.0000 1223692886.0000 1595261105.0000 1149901911.0000 1476112628.0000 1121604836.0000 1079248462.0000 1321827239.0000 1368934048.0000 trans-4-Hydroxy-L-proline 34029771.0000 42228954.0000 37789900.0000 19030828.0000 24760397.0000 26652682.0000 32863141.0000 69439770.0000 65960041.0000 77363894.0000 80972570.0000 14187344.0000 12837362.0000 13219540.0000 16513589.0000 81324606.0000 82209861.0000 85866015.0000 92828718.0000 50844759.0000 30126321.0000 40184262.0000 28140753.0000 24378241.0000 43931762.0000 51434580.0000 56813882.0000 UMP 26381569.0000 29903399.0000 30641166.0000 16996479.0000 15866758.0000 15521788.0000 17932108.0000 12321484.0000 11188218.0000 8450027.0000 10879896.0000 9984277.0000 9797194.0000 9326627.0000 10205741.0000 16772911.0000 16863298.0000 20178310.0000 21212486.0000 59255365.0000 39279064.0000 52246902.0000 54305045.0000 117827521.0000 101442142.0000 82235711.0000 75838160.0000 Uracil 2130714.0000 3334692.0000 3280145.0000 1149185.0000 1430524.0000 1016923.0000 1928184.0000 2089889.0000 2089907.0000 2944770.0000 3703643.0000 1586485.0000 1706035.0000 1969867.0000 1688239.0000 4926793.0000 5010431.0000 5161272.0000 5619760.0000 2726226.0000 2062470.0000 2706188.0000 2065939.0000 1527615.0000 2376956.0000 2001813.0000 2118598.0000 Valeryl-L-carnitine 99913456.0000 104846433.0000 108641315.0000 16775978.0000 18392816.0000 30466917.0000 35058367.0000 48258100.0000 46557330.0000 42584052.0000 51087756.0000 18394989.0000 19053738.0000 23588427.0000 27247582.0000 277545095.0000 366866981.0000 386951128.0000 400500549.0000 92229575.0000 67995230.0000 79936707.0000 66453953.0000 50923095.0000 52206982.0000 67016419.0000 75979411.0000 Xanthine 23858112.0000 39208197.0000 40020588.0000 28651448.0000 39540007.0000 25074480.0000 49069219.0000 24897249.0000 24687583.0000 33957820.0000 39020362.0000 31829348.0000 35475153.0000 45347844.0000 35730690.0000 39188707.0000 34878329.0000 36067158.0000 27352884.0000 55251954.0000 39331740.0000 58918474.0000 43680450.0000 29643481.0000 52100818.0000 42861929.0000 41221477.0000 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 2-Deoxyadenosine 5-monophosphate 2-Deoxyguanosine 5-triphosphate sodium salt hydrate 3-Hydroxy-3-methylglutaric acid 4-Imidazoleacrylic acid 5-Deoxy-5-(methylthio)adenosine 6-phosphogluconate ADP Allantoin AMP ATP Betaine B-Nicotinamide mononucleotide Butyryl-L-carnitine Citrate Citrulline CMP Creatine Cytidine Cytidine 5-diphosphocholine sodium salt dihydrate Decanoyl-L-carnitine D-Gluconic acid solution D-Glucosamine 6-phosphate sodium salt D-Glucose 6-phosphate potassium salt DL-2-Aminoadipic acid DL-lauroylcarnitine D-(+)-Trehalose dihydrate FAD GDP Glutathione (oxidized) Glutathione (reduced) GMP GTP Guanosine Hexanoyl-L-carnitine Hypotaurine IMP Inosine Isopentenyl pyrophosphate triammonium salt solution L-Acetylcarnitine L-Carnitine L-Carnosine L-Cystathionine L-Dihydroorotic acid L-Homoserine L-Kynurenine L-(-)-Malic acid L-Norvaline L-(+)-Tartaric acid Malonic acid Myristoyl-L-carnitine N-acetyl-neuraminic acid NAD NADH NADP NADPH N(alpha)-Acetyl-L-asparagine Nicotinamide Nicotinate N-Methyl-D-aspartic acid Oleoyl-L-carnitine Palmitoyl-L-carnitine Phosphocreatine Propionyl-L-carnitine Pyridoxine Ribose 5-phosphate S-(5-Adenosyl)-L-methionine p-toluenesulfonate salt Succinic acid Taurine trans-4-Hydroxy-L-proline UMP Uracil Valeryl-L-carnitine Xanthine METABOLITES_END #END