#METABOLOMICS WORKBENCH gmcgivney_20250506_102226 DATATRACK_ID:5889 STUDY_ID:ST003926 ANALYSIS_ID:AN006448 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 1 μl of derivatized sample was injected into a Trace 1300 GC (Thermo) fitted CH:CHROMATOGRAPHY_SUMMARY with a TraceGold TG-5SilMS column (Thermo) operating under the following CH:CHROMATOGRAPHY_SUMMARY conditions: split ratio = 20:1, split flow = 24 μl/minute, purge flow = 5 CH:CHROMATOGRAPHY_SUMMARY ml/minute, carrier mode = Constant Flow, and carrier flow rate = 1.2 ml/minute. CH:CHROMATOGRAPHY_SUMMARY The GC oven temperature gradient was as follows: 80°C for 3 minutes, increasing CH:CHROMATOGRAPHY_SUMMARY at a rate of 20°C/minute to 280°C, and holding at a temperature at 280°C for CH:CHROMATOGRAPHY_SUMMARY 8 minutes. CH:METHODS_FILENAME GC_LC-MS_analysis_protocol.txt CH:INSTRUMENT_NAME Thermo Trace GC 1300 CH:COLUMN_NAME TraceGold TG-5SilMS (30m x 0.25mm, 0.25um) CH:COLUMN_TEMPERATURE The GC oven temperature gradient was as follows: 80°C for 3 minutes, increasing CH:COLUMN_TEMPERATURE at a rate of 20°C/minute to 280°C, and holding at a temperature at 280°C for CH:COLUMN_TEMPERATURE 8 minutes. CH:FLOW_GRADIENT NA CH:FLOW_RATE 1.2 mL/min CH:SOLVENT_A NA CH:SOLVENT_B NA CH:CHROMATOGRAPHY_TYPE GC #ANALYSIS AN:ANALYSIS_TYPE MS AN:ANALYSIS_PROTOCOL_FILE GC_LC-MS_analysis_protocol.txt #MS MS:INSTRUMENT_NAME Thermo ISQ 7000 MS:INSTRUMENT_TYPE Single quadrupole MS:MS_TYPE EI MS:MS_COMMENTS Ion detection was performed by an ISQ 7000 mass spectrometer (Thermo) operated MS:MS_COMMENTS from 3.90 to 21.00 minutes in EI mode (-70eV) using select ion monitoring (SIM). MS:ION_MODE POSITIVE #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS Intensity MS_METABOLITE_DATA_START Samples GC_QC1 GC_QC2 GC_QC3 GC_QC4 GC_QC5 GC_QC6 Nf1.Cdkn2a.1.1.GC Nf1.Cdkn2a.1.2.GC Nf1.Cdkn2a.1.3.GC Nf1.Cdkn2a.1.4.GC Nf1.Cdkn2a.2.1.GC Nf1.Cdkn2a.2.2.GC Nf1.Cdkn2a.2.3.GC Nf1.Cdkn2a.2.4.GC Nf1.Cdkn2a.3.1.GC Nf1.Cdkn2a.3.2.GC Nf1.Cdkn2a.3.3.GC Nf1.Cdkn2a.3.4.GC Nf1.p53.1.1.GC Nf1.p53.1.2.GC Nf1.p53.1.3.GC Nf1.p53.1.4.GC Nf1.p53.2.1.GC Nf1.p53.2.2.GC Nf1.p53.2.3.GC Nf1.p53.2.4.GC Nf1.p53.3.1.GC Nf1.p53.3.2.GC Nf1.p53.3.3.GC Nf1.p53.3.4.GC 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:- 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-Hydroxybutyrate 13085719.6300 12997659.0800 13500008.4300 13367360.2500 13608795.9300 13042353.9700 6554462.6260 5817133.8610 5322445.7980 6619461.1290 6920402.6550 6668711.8790 6881711.7100 6777465.1240 11273883.3300 8170983.1640 8965312.0270 8482698.7970 7665443.0830 7026132.7640 8485785.9150 7617369.4820 6488108.3480 6379579.9180 6354482.3160 6167621.6110 7672762.5750 8238316.7890 7693181.8780 7735267.9850 2-Hydroxyglutarate 117548.7563 110671.1282 118229.2054 115323.0511 120644.4863 118291.8685 38351.2110 38965.8393 34911.6463 39476.6459 51300.4284 56022.3897 58308.2739 57267.5456 163309.3527 165689.9001 169510.8759 167631.0145 90094.0176 85800.0522 123303.7359 85556.4906 158217.9838 179826.6482 115953.1521 110148.1164 207061.7666 232185.2582 175003.4311 201181.2083 2-Oxoadipate 144614.2171 145508.4417 155435.9821 156059.7758 153306.9257 151441.8162 80047.8525 86103.7098 74762.2823 97575.4526 96419.7017 106279.1605 98805.6852 110907.3043 238617.4843 263742.3942 284004.1878 237484.3529 184035.7509 177570.1193 246629.4647 185708.0660 95819.1613 96451.3177 82437.5283 90712.6473 254456.5740 249918.0298 187367.1545 187215.0629 Aconitate 6099.9255 6255.7075 7790.3512 5067.1371 5238.8616 4172.8682 3602.7820 4144.6012 2784.3511 4002.2674 4201.5249 8393.4710 5842.7818 5442.8452 5445.5475 6500.9463 8804.4603 8090.0153 5946.1101 5492.3491 7281.5333 5128.4897 5269.9414 6681.5561 3541.1309 5357.6949 8893.0479 9354.9932 5731.1206 8023.1599 Adenine 220240.0074 228729.7640 238873.3372 243122.0151 257495.4837 252536.5694 97787.7054 104212.5331 85918.6952 130458.8377 163657.1364 147480.8272 146211.8029 173598.3587 277215.9192 300062.3427 320483.9384 228477.7509 214269.0001 170075.7897 228908.1015 205190.3366 228956.5486 277224.6888 196327.4354 175746.8846 283094.4084 386771.0307 283725.4068 261238.5881 Adenosine 18626.2831 17644.1697 18373.7447 16936.9207 18591.9606 17423.7456 10698.5919 10546.1404 10224.5600 11937.2724 8183.5774 9017.3917 9477.2092 10288.7194 31940.8353 33519.0842 42066.8675 28836.2673 21425.7587 20258.1535 32872.6127 22572.8731 12448.0258 10391.1824 14543.2613 14964.8626 30171.4428 23433.0315 18655.9447 23082.1039 Alanine 164219252.9000 162085104.0000 162060419.5000 160645680.1000 162263187.6000 155421090.4000 42267430.0500 43810127.5300 41475999.6300 61795194.5600 55643851.0700 60088147.6800 68439056.8400 78888347.4500 174581304.0000 186785515.9000 240017742.3000 177554487.8000 205514187.9000 153185876.3000 235940459.0000 150127924.7000 87204384.7700 88873540.2800 97592459.2100 106351612.5000 270632779.5000 299705695.3000 229715217.4000 259230384.6000 alpha-Keto-beta-Methylvalerate (KMV) 93418.4596 92715.2706 94819.6266 95441.8154 99061.2552 94927.4809 24255.7206 25142.1594 19460.1779 34574.6672 19589.9414 23319.2344 29468.1450 28632.1439 131177.4095 179371.0181 159026.3873 130134.6083 86044.2767 66876.0832 109654.6384 65492.2068 50396.6482 69504.6557 50058.4518 53205.5753 189493.8029 279641.2205 155742.9649 183864.3579 alpha-Ketoglutarate 861598.9166 902552.4381 904100.0020 879229.2963 925575.5265 902463.6888 243503.2649 268489.7313 176993.6259 320480.5793 445991.5245 442036.7631 384281.0935 416755.2368 1943645.9460 2001820.4760 2032584.5340 1996858.8060 695205.8511 770367.5928 1053454.2680 784254.5521 695727.2763 986591.0122 478694.3979 407459.5206 1679097.3090 2036665.5680 1429964.8790 1602570.1460 alpha-Ketoisocaproate (KIC) 151743.5851 152675.8770 159125.2427 147985.3301 157050.0897 153266.3530 54047.5204 60735.3504 48046.3461 71115.6305 46402.4339 54998.2941 65944.8772 62748.6144 212926.8927 272270.3056 244794.5389 207025.9931 159664.3274 119700.9067 194509.0626 117119.6351 95331.1043 128777.8374 99338.3402 99733.4351 298719.7835 420170.6532 240041.6899 292000.5800 alpha-Ketoisovalerate (KIV) 20593.9683 20907.4791 22276.9897 21400.1131 24118.5670 22235.2321 3093.3480 3634.3569 2211.5927 5584.9975 3306.5293 2926.0829 4340.9432 4825.0531 27866.9658 38052.0056 34636.7671 25843.6544 20223.1807 13635.3429 22362.8005 12711.2422 6148.7497 9301.7170 7612.7503 8154.1744 54379.8553 74670.3027 43716.6537 52904.8563 Aminoadipate 169487.3953 171264.1788 174426.6523 173436.8608 179842.8325 174799.8423 44419.0346 42750.7555 39901.8616 67783.4848 84123.3316 70940.1662 93264.7189 104663.3237 209086.2096 211135.1697 198691.7206 187133.5677 142351.5776 104463.6000 140815.4553 112727.4542 174988.4887 138677.2240 158665.9400 133860.6352 194866.7812 147620.5422 106446.4676 93657.8064 Asparagine 548666.9254 448799.9123 443850.2153 451119.6784 471440.5333 457247.8741 75957.6152 80607.5209 70493.7582 114871.3412 81469.0525 102999.2095 118249.9510 138696.2164 341933.6799 328467.3360 358220.9463 389592.5653 1433150.5620 879343.4472 1612276.9630 909414.5374 381424.8581 328273.4447 403436.5953 454172.2425 556978.0039 504095.2500 330651.4210 344882.3955 Aspartate 24435118.8100 23418614.3900 23647961.0600 23534145.0800 23894046.7000 22997140.1600 13540342.6000 13935138.2100 15545871.7900 21690367.9100 20105037.6600 20836834.8800 26945667.8400 33759549.7200 34785778.8400 36854095.7900 37597164.6300 33225446.7900 28120747.1800 21515569.8100 36754605.6000 21052440.0600 14417831.3300 16385065.2400 16720810.9400 18937788.9300 17565953.0100 12065433.9000 8338552.8490 7603170.0310 beta-Alanine 5096784.5110 5720062.9830 9592459.9820 10431534.8100 11140025.4600 10647562.3500 6423976.6450 6213525.6460 7642336.2870 9491820.9040 7557659.0590 7611629.0630 9844994.5950 11498260.4100 25354863.7800 25949554.3600 26805542.5300 26435106.7100 11317972.8500 7962764.5030 12389415.4100 7970425.2480 8330603.9470 7898762.6590 9088212.1350 10013383.8700 12377974.2900 14528855.6100 11988556.1700 13374908.1800 beta-Hydroxy beta-Methylbutyric acid (HMB) 1001312.7350 995882.6311 1012895.9910 1007036.8600 996761.1591 983349.4144 1159001.5510 950720.1600 943992.8796 965851.0893 873799.1308 1007959.0930 913941.2006 975300.3506 1038368.7310 1041174.3650 1176724.2320 998531.1364 913825.6218 975506.5476 1259997.8170 901569.9592 1184712.3060 1049227.2490 984600.3944 947649.6931 968776.9031 1049125.1180 850847.0403 957125.1282 beta-Hydroxybutyrate (3-Hydroxybutyrate) 102706.3378 101642.4931 105570.4033 105974.2118 108294.3745 106881.9378 35570.8510 40593.2668 32913.8027 50453.2615 41360.1222 45691.7442 52365.0211 54593.7210 131460.4920 156994.5814 174532.3709 136139.4950 77001.4063 64063.6756 89248.8921 61283.1282 51476.4104 63072.4833 47774.7106 53578.8184 185187.6064 214018.2966 157945.3599 216912.1371 Cholesterol 11182636.5400 11033323.0000 11342621.0900 11054670.2000 11530524.9700 11268133.3100 7505786.8290 6945819.7740 6168250.7810 7284435.5120 7364660.4640 7382191.1980 7358735.3660 7524655.8650 24830920.6000 22616782.4800 28682796.9300 24119014.7200 10051605.8000 9617411.3350 14210706.4500 9596564.0060 8436508.1410 8069677.4160 6962593.2210 7365784.3870 14583520.1100 13369153.5800 12312753.9900 13720774.5800 Cysteine 1728063.9200 1746821.0940 1787250.9600 1768316.1420 1778077.1960 1749673.9760 685986.2345 792379.9845 504585.4784 1038596.4580 777300.7240 1001105.5030 1275897.7190 1282353.9820 2429724.6400 4121929.5010 2891847.5980 2381397.8550 2187836.3100 1306728.7630 3142871.8070 1191036.1420 1349015.5020 2323011.5440 1539156.4860 1506386.8270 2363641.1560 4515183.8760 2164185.1290 2401795.7930 Dehydroepiandrosterone (DHEA) 13584.7453 17290.3905 12902.3331 12285.4914 18238.6452 12774.0850 12231.2359 9256.2393 12650.9541 10888.1591 11764.0918 10439.4118 8733.0090 10139.0255 14320.9349 15703.7265 17362.1502 12868.7430 11946.9861 11868.6970 15159.5590 8940.1976 11817.0452 10984.4215 11490.2546 10924.7562 12624.0108 17246.8373 11385.7241 13065.6260 Dihydroxyacetone phosphate (DHAP) 64742.4172 73587.4925 85844.3871 83838.8859 89257.7249 84414.1807 39089.7452 38492.7149 27147.5904 46582.2227 44980.0617 41483.8795 44744.0923 48513.4667 135357.6633 123613.1041 136874.9997 132623.1637 101649.3946 80561.1786 132556.6894 95348.5811 186239.0115 191620.7869 126795.9925 129848.7374 76494.4516 89877.2126 73620.7436 74842.5997 Fructose 1488513.7660 1487081.7430 1561967.6530 1542873.9250 1583149.3610 1512945.8900 665248.6031 701255.2151 614321.6392 1523263.9280 902552.6507 915314.2547 1346102.3760 1438239.2300 3572768.4890 4644091.7630 4149542.7070 3510557.5570 1465164.3790 716769.0519 1707076.5790 731377.3817 1284463.3630 1823999.3450 1386578.4520 1693703.6310 2423045.0500 4489415.9370 2382440.7190 2809945.7760 Fructose 6-phosphate 152637.0647 179929.1232 168831.3365 150069.4324 134920.3935 142508.3780 65741.0332 65240.7368 55496.9831 90883.1076 80368.4016 74631.2076 86142.5005 102155.7918 312216.8188 278644.8144 356263.8025 262286.4226 100865.5313 67134.1469 120204.2424 65668.7778 108430.0956 130700.5280 88680.0415 103704.4229 210354.9519 203970.7356 182158.5367 200613.9498 Fumarate 1340547.9710 1326730.8970 1360448.7530 1342271.9630 1423796.3180 1318627.7400 621281.7710 646043.6875 531622.4250 792690.4565 1182941.9140 1250927.5970 1270199.7280 1351511.8830 1925889.6850 1830684.0830 2095345.1200 1769957.9600 1202025.1100 1126911.7800 1601412.2250 1119935.1500 996142.7400 1200572.7640 884880.4555 839382.3730 1410477.3310 1562023.2010 1214465.8340 1288985.1250 Glucose 11770446.8700 11926967.5700 12826239.2500 12808461.2000 12827573.6600 12336271.0700 3811387.2490 4699133.6620 3321328.2690 13593425.2300 9649334.2340 5237248.4530 10729685.6100 9644400.9240 25240712.5900 40681525.5900 26430038.1800 24476750.4600 14107359.8900 6291135.8910 16652439.4300 5291865.4890 7828452.9430 17219522.6200 9030927.6330 11721045.9800 12292747.6600 28764642.4400 10653799.8500 11785958.0800 Glutamate 180633799.4000 177069027.4000 187193574.0000 187603016.5000 189113084.6000 178150243.3000 89705610.4000 91855319.1600 84332538.8300 123840671.4000 154915385.3000 147396030.0000 164848655.9000 181041297.2000 259418354.7000 265574050.0000 293415742.8000 265364506.0000 230252326.0000 193650649.5000 289434303.8000 191317244.4000 164747210.6000 177486983.8000 169355965.6000 170935703.4000 204618676.5000 225783655.1000 173166590.2000 180615802.0000 Glutamine 17019593.6700 16652993.7100 17416241.7400 17398580.8200 17947432.2900 17139962.5100 9550688.4050 10178670.3600 11281576.0700 15744057.1600 8017304.5000 9264606.2220 11917679.4200 13887310.8200 23740257.1000 27566945.3200 26189580.7500 24659568.6600 26120541.5900 15900893.9100 32987917.4500 16222413.1100 13244198.2200 14991493.2900 15928449.5400 20504394.0200 26567616.4000 33043102.9900 22310679.8900 23849272.4100 Glycerate 89135.5738 88689.3898 90367.1136 93126.1342 94592.9610 90389.4757 41784.7193 46637.7606 39946.2132 56197.1832 49840.5507 51566.5721 56706.7255 60907.2907 130438.7125 145283.3234 140340.8016 131273.9079 70121.2384 59795.2543 91420.6808 64481.0649 68677.6965 77643.3635 62665.0123 67138.6227 157881.3274 249082.9275 173864.9765 193550.9766 Glycerol 59131.3015 60203.9322 64913.3117 66191.9820 67208.0905 62673.4000 66735.9886 50433.4786 48522.6217 57472.2538 44299.8235 48318.3668 49336.4415 48300.4365 90159.3724 92565.9033 97470.0085 84802.0210 50116.9105 41883.8296 63360.8032 47966.3545 44064.1373 42671.1833 37544.8943 43081.4324 98523.7488 90952.7040 66480.8001 79787.1365 Glycerol Monolaurate (GML) 806099.6934 851899.2399 895386.2310 903334.5833 935248.0846 881616.5480 434719.7287 350823.4339 280012.5212 426903.2010 426908.2075 456079.8603 382387.9790 454868.0771 2125548.9070 1458437.7350 2282853.4200 1469903.1720 548331.8649 478227.4322 963433.6813 469581.6640 456163.9376 452106.4609 363361.1417 418205.5847 933085.2183 865036.5258 771752.4591 1019021.9970 Glycine 180717112.1000 202507357.5000 262775136.6000 260401179.1000 266369754.2000 254848300.9000 77759322.9600 88150001.7900 70299966.3000 109326061.5000 93839357.1200 98092187.7600 109714272.8000 120132055.1000 365132885.8000 390470137.9000 402477295.2000 377388374.1000 389649399.3000 307283480.2000 468175860.1000 330394688.4000 131300190.9000 130636847.4000 128527143.1000 136457501.4000 532276647.0000 606996364.0000 459725656.0000 537598925.8000 Heptadecanoic acid 63685.7285 76871.6107 87905.0130 85443.4477 99086.4556 91173.0374 89209.6243 63328.9995 58530.7208 64513.8446 72271.8247 75186.4442 72171.1106 74476.9490 107260.4146 112343.1150 126824.3283 106728.1051 74181.0439 68933.4658 106424.0689 68587.3180 67247.6241 63935.8493 57689.8447 62092.7291 90375.3640 107329.1478 74396.4492 73733.8888 Histidine 1054227.3760 1347143.1050 1484799.9090 1441222.8140 1342209.8400 1392372.7690 692280.7332 628662.0119 664278.4459 1001069.1540 499785.9547 535035.3907 713683.2471 740060.9991 1095006.9730 1695449.4780 1145143.0600 925299.4932 1802650.6140 1065937.7280 2296297.2350 1016059.0500 921171.7037 984543.6936 841399.2827 1000261.7620 1108775.5110 2115358.2020 928731.8792 1053168.7840 Homocysteine 71808.2211 72542.5141 77133.2830 72959.5953 74731.3659 66898.4421 33029.9342 32536.7962 27736.6751 33800.0202 36065.8608 40813.9524 41790.3330 50138.7353 109858.8967 129391.2211 145336.6822 91701.2711 114648.3005 79680.1586 153379.0250 85423.9189 63075.4872 63721.9878 65189.4448 67314.7635 101253.1679 132250.6202 88076.9494 83927.7984 Hypoxanthine 31905.5538 32950.5148 32573.1094 31947.0165 35819.8879 32398.0444 31596.0142 39356.5607 26118.1668 43059.0277 25343.0894 29663.8630 31902.4811 36152.3474 29847.4738 28624.9498 32460.2070 26815.3468 52891.8392 34465.7335 70553.1441 46844.7640 49103.2614 51569.2754 46890.5342 44533.8147 38711.5934 36311.3586 31072.6315 34212.7709 Inotisol 90468330.3700 89708900.3200 95456417.7300 96018210.8200 97907119.2000 93840917.8800 34947978.5500 35871565.5900 39836995.6400 58143709.9800 69755589.7200 69764232.0900 80298800.0600 96134029.3400 174542388.9000 177871704.3000 192953473.4000 178297796.3000 87309937.3300 61942205.2400 103299149.4000 67921214.2900 53949341.1200 41858416.8500 54550999.0000 63921955.2300 147730984.9000 161454014.8000 132859454.0000 152001907.3000 Isocitrate 137785.8873 131509.6948 132520.3308 135639.6690 143562.1702 133955.4092 66217.9098 73186.4559 51982.7945 74619.8069 107272.0409 119588.7220 112273.0675 113273.1599 230842.9613 236521.0497 295308.5530 219805.0675 108991.6053 107162.1973 138866.4990 105703.5927 110284.1947 137432.5854 83069.3559 77573.0337 210069.7860 242432.2727 203283.4921 232869.2197 Isoleucine 24487434.7700 23729150.8800 24877332.5000 23957448.6000 24834304.8000 24461046.9200 10229438.3000 10896741.9400 10212544.0700 16398228.4500 10675526.4100 11611067.8400 16320906.3300 17696786.9100 30937431.5000 39409285.9200 33859803.3000 31051581.5400 33484604.4000 20019389.6900 38734861.7100 20755866.7500 13465389.6900 16909062.7400 14667855.9800 16916583.2500 34045398.0100 41567684.4000 27207211.6300 30867708.7800 Lactate 180825331.6000 183760445.7000 187953684.0000 183080284.1000 186147594.7000 180231903.8000 44126296.7300 54105187.9500 32016236.9300 66207655.6300 40221390.3800 44455931.8700 55014176.9900 63270269.5700 275974197.5000 353039110.3000 329168317.4000 272343547.3000 101712241.2000 74574764.3400 110888660.1000 75956567.4000 71938975.0900 98449804.8100 71109698.4300 77441944.8300 418472149.3000 503118392.7000 320070106.6000 444369276.3000 Lauric acid 500558.3864 524202.0676 512212.3499 507471.9526 516992.5067 512016.1159 310265.4054 321138.1007 278098.6310 382349.3534 339332.3616 343459.4158 340104.6801 384493.5253 766813.3197 849474.2353 909859.3361 759688.6043 483992.8897 482862.8768 633377.2533 527872.7666 323034.5177 336642.7437 298897.1612 311724.4442 859250.6381 907720.4619 639979.7674 685937.4798 Leucine 31209831.1500 30366508.9400 31497306.1300 30793180.8200 31395282.5300 30953380.4400 13418028.7700 14098748.8600 13437294.0300 21054682.7800 13854348.5400 15085594.3100 20847886.3500 22955954.4400 37423465.9600 47767776.3600 40247033.8700 37864702.0700 42078752.3300 25788225.3500 48491081.4100 26738178.7000 18126540.1700 21786797.3300 19343377.1200 22001605.8000 42581666.6400 52253527.3700 34156534.8200 37825231.5500 Linolenic acid 2005.2795 3129.7534 2574.8956 3764.6058 3340.4517 2423.1309 1552.6804 1967.9136 754.7195 1467.1733 1901.0231 2300.8876 1735.3229 2085.7288 6248.2337 5222.1129 7172.0803 6220.0584 3257.5469 1963.6190 3649.7033 2153.1950 2385.8180 2518.5450 1799.3767 1734.1322 3416.1104 4694.7647 3431.8584 4149.0682 Lysine 4738026.5280 4985577.1470 5492490.2120 5333654.6140 5496873.6570 5370909.8790 4258062.0430 4364541.2760 4234477.0550 6424362.5910 3291806.5760 3589685.8020 4529618.0290 5077589.6770 6029652.1310 8815115.0530 7695831.4950 5716667.4880 8505617.4940 5421879.0950 9770190.3440 5444376.3680 5826264.0010 7077635.6620 5661296.3440 6731626.3850 6272468.3910 9269982.6450 5222859.9130 5894992.9580 Malate 1388991.1830 1346041.4800 1394140.9090 1397902.6860 1432002.1330 1374438.3580 554934.0266 549516.1393 482685.3934 738143.6787 1305106.4070 1337435.9350 1391718.4130 1525842.2970 1806390.8330 1803484.3940 2154450.1410 1747500.5590 1235004.6590 1191963.1170 1836795.9750 1173667.1200 926903.3332 1123377.2890 786766.5443 734422.7184 1353052.6080 1482196.0230 1128929.0580 1251199.1530 Methionine 5756971.3820 5759782.8660 5999133.3180 6056097.1920 6114401.7860 5970485.6860 2680145.5780 2829037.0340 3202379.9420 4482889.9030 2931881.0780 3127917.1950 4076896.7520 4606459.4100 6984333.9690 8565198.7130 7721034.5190 7146518.6820 8860220.6550 5477518.1730 10423930.4800 5646458.5690 3741354.6720 4592737.9220 4584324.9870 5389627.0960 8565309.8720 11272862.3600 7617569.7260 8061422.1740 Myristic acid 621009.3172 716545.6590 756549.6610 735898.7091 885632.8871 782491.1733 659917.1030 605556.3926 558290.3729 639027.3381 622247.3163 646414.6600 645141.3328 670791.0332 966913.4381 986041.2652 1143410.1020 897513.8473 688912.3773 626909.9841 838589.5510 673172.6297 420117.9762 447700.8659 406970.3741 430990.0382 1224632.1320 1317326.6090 917045.0619 951537.8131 N-acetylglutamate 46839.4751 45292.3761 45766.9135 47624.1253 56090.6600 50031.2006 22849.0586 24365.2120 20188.1286 29241.2322 32503.8145 35494.3043 34286.4958 43893.0339 99015.5764 97219.2378 96557.9603 104129.1011 65075.1485 46837.6243 94430.0653 50262.4476 22427.4721 25710.8171 17778.1983 20712.1678 67002.4737 78995.7026 56074.5000 60930.2257 N-Acetyltyrosine 5637596.1960 5745371.0120 6154363.3290 6071258.2510 6029991.0020 5916865.8160 2170418.4390 2206422.1940 2553733.7370 3679345.2370 4430441.0980 4415871.8290 5087149.6950 6064178.9650 11033586.1400 11486732.2900 12310629.3600 11636172.0500 5511806.0510 4009413.3770 6615723.4790 4231577.2700 3469191.8410 2651106.5230 3477903.7130 4089508.5830 9634631.2630 10240329.5700 8388411.9420 9527399.7080 Oleic acid 1947846.8100 2705592.8580 2649142.1530 3220452.2290 4336998.0980 3614845.6010 1943295.7200 1856051.5970 1799971.4870 1981891.6950 1716139.2140 1702252.1280 1636056.0480 1728633.9890 6008226.7330 6008279.9430 7500454.0800 6408463.5030 2552795.6390 2165713.4720 3065959.3720 2162973.3910 1995825.9920 1967994.2470 1557984.4960 1675171.8200 3710209.9670 4206057.4110 3838335.7810 4138640.3260 O-Phosphoserine 124975.9150 148232.0219 158993.6681 171426.6942 179028.8096 166766.2035 170157.1454 172278.2551 177940.2360 288673.7604 249900.0748 240516.4961 322728.5192 383273.5899 116895.4126 111385.1493 114192.6512 120706.7123 292199.7157 200436.6370 409691.5792 200371.6705 35821.7675 61058.2155 76252.1904 97350.5879 96887.8761 82839.1370 58279.5695 39165.0110 Palmitate 7454887.8900 8781044.4920 9095569.8960 9339864.5780 11749942.7500 10178802.0700 6315372.2530 6040432.3670 5454694.7260 5858419.4850 5979016.3830 6584993.4580 6453503.3150 6301198.0460 12288950.3700 12965015.1500 14409780.0400 12518455.6300 6844959.9390 6371839.1770 8957862.2770 6461834.4410 5932585.9650 5969952.2220 5035718.3740 5453758.6920 8563927.7980 9416603.8890 8177010.5530 8257534.6390 Phenylalanine 8130612.3230 8059047.9090 8541681.3020 8080936.8420 8307166.9280 8120939.8870 3633133.1540 3721787.2070 3667547.4030 5787650.4890 3833210.3310 4139744.6020 5194242.0490 5603726.5110 9642390.0380 11879305.0800 10933118.6600 9862818.0040 11883501.2500 7637713.1450 14212586.4400 7657322.2240 5216013.2090 6213688.9130 5560327.3150 6435480.8850 11935305.3100 15514547.2500 10204973.7800 11539633.2800 Phosphoenolpyruvate 18268647.6000 18359515.4800 20236497.5300 20761162.6100 23565748.9300 22441325.7300 11283725.2000 11737250.2200 10433511.9000 13531767.7300 16371420.3500 17529698.0900 17389031.2300 20243847.4800 35619650.3300 33804492.7900 41767314.9500 35292193.3000 24112393.1100 22071214.6100 31955717.3300 22692571.5300 14597901.8100 16711553.0900 13321944.9200 14803633.5400 31516827.4300 29087640.7300 27111125.2400 28912157.4700 Proline 52508919.8100 48966679.9500 49825934.1600 46099083.5600 48128335.5500 47885710.7100 6005404.6550 5871320.9190 6677302.8690 8380410.6260 6131083.1600 6321862.7230 8210930.3650 9308523.4120 23389474.4600 26178643.8300 29499196.5100 23857212.6500 81560030.7300 46500252.9600 86157591.3700 48449882.3400 14525033.2300 12776182.3100 20510372.5100 24573585.4100 130902820.5000 126411665.3000 101218287.6000 140897918.3000 Pterin 66212.0593 65999.2562 72436.6581 71743.7470 73973.0203 71389.1481 8127.3212 9568.3719 8927.8545 22907.1216 19529.0086 15816.7756 27841.3326 28648.3973 71033.6039 80167.9135 77066.3218 72659.2824 84232.4321 42549.7866 94265.8839 48309.2930 43278.4345 43918.0647 35907.6289 42762.5436 149720.8942 128483.7649 89861.7141 98090.3038 Pyruvate 2778283.7320 2818851.4610 2865652.6530 2863494.6980 2896866.4760 2810265.6180 1014664.6860 1402485.1430 1003994.0180 2008625.5170 1187731.8660 1314754.3580 1898625.3240 2063658.6950 5759836.8850 8065374.1560 6228073.1720 5723547.7630 2450145.8990 1413229.1510 2877220.1440 1567437.6510 1293094.0090 2427878.0210 1461062.6080 1710987.6270 4169302.4580 5119546.5070 2761017.2430 2945995.5950 Ribose 54578.5279 59070.8047 58959.7314 55698.1640 56711.7286 55945.6868 18350.8073 18745.8216 13138.1154 29344.8652 23093.4213 23719.6098 31752.5114 29316.4254 95885.0429 115691.3873 101231.1555 94646.7987 68807.9504 49943.5507 85821.4690 47730.2298 50665.8688 61301.0883 44590.7399 48859.6084 97538.8041 138624.3638 83065.6891 102987.9412 Sedoheptulose 7-phosphate 10365.8171 11844.6247 12941.3926 14160.1438 15566.7477 14548.5077 3746.5525 3032.0153 2444.9638 4903.8456 4890.5917 3269.7052 4458.4986 3835.4418 12970.0982 14825.0903 14262.6897 13572.4319 19206.0763 15846.4251 28277.2231 14739.6863 12745.0528 17306.8335 11879.0708 12023.1046 38599.4415 51684.0274 33093.9359 41764.6501 Serine 30881907.8400 29463429.5200 32304855.0200 29568931.8700 30766204.7800 30439577.7300 15411941.6900 17622591.6900 18343701.5800 22951688.7900 18335401.6200 17264857.9900 25121118.9400 28382122.1400 22081122.4400 27901327.6900 21977984.3400 22406111.6700 61502663.5400 35156877.7200 66983516.7400 43254422.6900 19252787.8200 21215740.5300 23008815.6700 26566762.6000 36875330.3100 42723274.0000 30384576.9300 27844934.3200 Sphingosine 65955.5989 75372.6472 76023.9916 67861.5803 73085.8555 67180.3165 57314.5184 57528.2824 57280.1046 64353.8328 41452.4833 48800.0564 42323.9941 55220.6658 153844.7714 157713.3922 164282.4106 144354.3621 47928.0042 37768.7336 66215.4404 35171.6399 63512.3613 50733.0369 47960.0324 58589.2566 78018.9382 77484.5414 64873.5875 65256.3555 Stearate 4967104.9540 5403877.8420 5984565.0720 6293045.6820 7037221.6710 6609824.0150 3593785.3790 3618230.8710 3433614.2130 3628608.2160 3213070.9370 3738700.6390 3337408.9920 3123290.0530 6318347.6890 6734965.3390 6757196.5570 6021168.4730 3543255.8110 3832241.6870 5505527.5730 3796182.8620 3865615.1880 3325836.7560 2927219.2530 3106581.9530 4300075.1380 4693444.8050 4466437.4280 4314556.7920 Succinate 1167327.4530 1344399.0150 1746658.5530 1745240.1910 1821907.1500 1730334.0720 534176.4828 591050.9866 462292.8559 736297.9516 620357.2582 645835.1565 738790.4065 813839.3316 2537024.9070 2690109.2620 2758080.9600 2566512.4640 2745100.9430 2102281.3940 3231595.8170 2258168.5750 878670.1485 875641.3010 874628.6308 917318.9576 3644042.7280 4226801.1200 3142145.0900 3725976.2880 Threonine 32092609.3900 31557015.6000 33387736.8400 32105903.1000 32841448.3300 32421893.5600 11656145.1500 12568246.1700 13994951.1700 17752725.3200 13611131.7000 12960384.4900 18490133.1500 20222964.8300 39370773.3200 47085003.9100 52711883.3800 40095964.3100 49057339.2100 27942841.4500 52247246.9900 32359352.4600 17623880.2500 18850235.5200 22560373.7600 25966065.6900 47210565.8800 49033036.9700 38314290.0900 44169964.4400 Thymine 20618.5761 39469.8554 57105.1027 25250.6595 64848.2085 69026.8045 19214.6722 45417.3124 19814.2177 18797.2396 28177.2523 26880.2887 19842.8764 19436.4841 31678.8699 28241.7451 57082.1828 33806.4016 32179.5789 22284.5721 36178.9998 21576.8704 40339.0011 57727.7056 21823.2625 24730.5187 59582.6853 32433.2894 28167.6238 33919.4814 Tryptophan 4576562.9850 4697626.4250 4794742.3260 4801678.1090 4884863.4840 4745337.0250 1944943.6420 1889898.0070 1876729.1700 3213185.3310 1939242.4510 2195799.3580 2816486.8140 3100317.5040 6151864.1550 7722377.2450 6987035.2690 6363248.2480 6685813.6450 4266082.5230 8516782.0550 4195887.6460 2944217.1860 3630123.5360 3061361.5680 3460416.2390 6952995.6790 9235305.7660 5821794.5820 6348140.6040 Tyrosine 1492867.6760 1520595.3710 1605336.3680 1637112.4440 1649961.8980 1588234.5410 568768.8434 598187.0483 592246.6642 1029851.5530 593477.0845 663950.2396 907278.8614 1008480.8850 2072013.8650 2661036.8890 2420926.6750 2073946.0940 2332022.6410 1391542.5930 2903921.3960 1425733.1660 874606.2847 1137941.4360 999877.0491 1179228.0700 2587786.2400 3393695.2170 2138853.4140 2461110.2890 Urea 7729811.8910 7603136.2100 7868785.5160 7810114.0120 8137023.3940 7915303.0590 5436047.9720 6478100.7410 4908040.2150 6911708.1750 3475797.5670 3430343.9250 3859800.0590 5371692.6270 7883710.8790 10545394.8200 9262082.7550 7730361.5140 8228995.6340 5532051.6280 7435151.1120 6512814.3440 5493477.8200 6283702.9090 4681793.6120 5303363.0890 28729569.4500 14618890.8500 8009506.4470 7220668.3330 Valine 25989404.1700 26090515.4500 26444358.6700 26259659.7600 26178239.7400 25545229.8200 10962960.5700 10832229.8900 10374982.2100 17887505.8600 10592670.3900 12617990.2100 16376059.9700 19160662.3100 34230241.2400 42269537.0500 35799050.4200 34604796.1000 36433323.0700 23434654.4800 43794640.5900 21630958.7800 15306250.6500 19381680.3000 16068314.4200 17592119.8400 36268164.0500 47988888.1300 30864218.9600 34101800.2900 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-Hydroxybutyrate 2-Hydroxyglutarate 2-Oxoadipate Aconitate Adenine Adenosine Alanine alpha-Keto-beta-Methylvalerate (KMV) alpha-Ketoglutarate alpha-Ketoisocaproate (KIC) alpha-Ketoisovalerate (KIV) Aminoadipate Asparagine Aspartate beta-Alanine beta-Hydroxy beta-Methylbutyric acid (HMB) beta-Hydroxybutyrate (3-Hydroxybutyrate) Cholesterol Cysteine Dehydroepiandrosterone (DHEA) Dihydroxyacetone phosphate (DHAP) Fructose Fructose 6-phosphate Fumarate Glucose Glutamate Glutamine Glycerate Glycerol Glycerol Monolaurate (GML) Glycine Heptadecanoic acid Histidine Homocysteine Hypoxanthine Inotisol Isocitrate Isoleucine Lactate Lauric acid Leucine Linolenic acid Lysine Malate Methionine Myristic acid N-acetylglutamate N-Acetyltyrosine Oleic acid O-Phosphoserine Palmitate Phenylalanine Phosphoenolpyruvate Proline Pterin Pyruvate Ribose Sedoheptulose 7-phosphate Serine Sphingosine Stearate Succinate Threonine Thymine Tryptophan Tyrosine Urea Valine METABOLITES_END #END