#METABOLOMICS WORKBENCH manjari1_20240713_133923 DATATRACK_ID:5007 STUDY_ID:ST003388 ANALYSIS_ID:AN005555 PROJECT_ID:PR002099 VERSION 1 CREATED_ON August 7, 2024, 8:23 pm #PROJECT PR:PROJECT_TITLE PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2 PR:PROJECT_TYPE Lipidomics of Human lung epithelial cells PR:PROJECT_SUMMARY In this study, we calculated the peak area abundance of phosphoinositide lipids PR:PROJECT_SUMMARY in SARS-Cov2 infected, uninfected and dual lipid kinase (PIP4K2C and PIKfyve) PR:PROJECT_SUMMARY inhibitor treated (RMC-113,) (PI, PIP, PIP2 and PIP3) Human lung epithelial PR:PROJECT_SUMMARY cells. We discovered PIP4K2C’s (Phosphatidylinositol-5-phosphate 4-kinase, PR:PROJECT_SUMMARY type II, gamma) roles in SARS-CoV-2 entry, RNA replication, and assembly/egress, PR:PROJECT_SUMMARY validating it as a druggable antiviral target. Integrating proteomics, PR:PROJECT_SUMMARY single-cell transcriptomics, and functional assays revealed that PIP4K2C binds PR:PROJECT_SUMMARY SARS-CoV-2 proteins and regulates virus-induced impairment of autophagic flux. PR:PROJECT_SUMMARY Reversing this autophagic flux impairment is a mechanism of antiviral action of PR:PROJECT_SUMMARY RMC-113(dual lipid kinase inhibitor (PIP4K2C and PIKfyve). These findings reveal PR:PROJECT_SUMMARY virus-induced autophagy regulation via PIP4K2C, an understudied kinase, and PR:PROJECT_SUMMARY propose dual inhibition of PIP4K2C (Phosphatidylinositol-5-phosphate 4-kinase, PR:PROJECT_SUMMARY type II, gamma) and PIKfyve (a FYVE finger-containing phosphoinositide kinase) PR:PROJECT_SUMMARY as a candidate strategy to combat emerging viruses. PR:INSTITUTE Stanford School of Medicine PR:DEPARTMENT medicine PR:LABORATORY Einav lab PR:LAST_NAME mishra PR:FIRST_NAME manjari PR:ADDRESS 3332 middlefield road PR:EMAIL manjari1@stanford.edu PR:PHONE 6503849709 PR:FUNDING_SOURCE NIH PR:PUBLICATIONS https://www.biorxiv.org/content/10.1101/2024.04.15.589676v1.full PR:CONTRIBUTORS Marwah Karim, Manjari Mishra, Chieh-Wen Lo, Sirle Saul, Halise Busra Cagirici, PR:CONTRIBUTORS Do Hoang Nhu Tran, Aditi Agrawal, Luca Ghita, Amrita Ojha, Michael P. East, PR:CONTRIBUTORS Karen Anbro Gammeltoft, Malaya Kumar Sahoo, Gary L. Johnson, Soumita Das, Dirk PR:CONTRIBUTORS Jochmans, Courtney A. Cohen, Judith Gottwein, John Dye, Norma Neff, Benjamin A. PR:CONTRIBUTORS Pinsky, Tuomo Laitinen, Tatu Pantsar, Antti Poso, Fabio Zanini, Steven De PR:CONTRIBUTORS Jonghe, Christopher R M Asquith, Shirit Einav #STUDY ST:STUDY_TITLE RMC-113 (dual-lipid kinase inhibitor, PIKfyve, PIP4K2C) alters the ST:STUDY_TITLE phosphoinositide regioisomer signature by advanced lipidomics analysis ST:STUDY_TYPE lipid stduy of phosphoinostide lipids ST:STUDY_SUMMARY To determine whether RMC-113 treatment impacts phosphoinositide abundance, lipid ST:STUDY_SUMMARY extracts derived from uninfected and SARS-CoV-2-infected A549-ACE2 cells were ST:STUDY_SUMMARY subject to lipidomic analysis. Employing Phosphoinositide Regioisomer ST:STUDY_SUMMARY Measurement by Chiral column chromatography and Mass Spectrometry (PRMC-MS), we ST:STUDY_SUMMARY comprehensively profiled all eight PI classes and their acyl chain variants ST:STUDY_SUMMARY (defined by the carbon number and the saturation level). Seven of the eight ST:STUDY_SUMMARY phosphoinositide classes (except for PI(3,4)P2) were detected in all tested ST:STUDY_SUMMARY conditions. Upon SARS-CoV-2 infection, the abundance of multiple PI classes was ST:STUDY_SUMMARY increased relative to uninfected samples, albeit with some variability across ST:STUDY_SUMMARY independent experiment. This increase was most pronounced with the abundant acyl ST:STUDY_SUMMARY chain (38:4), yet a similar trend was observed with other acyl chains. Notably, ST:STUDY_SUMMARY RMC-113 treatment in infected cells caused a 1.5-2-fold increase in the ST:STUDY_SUMMARY abundance of PI3P and PI5P—the substrates of PIKfyve and PIP4K2C, ST:STUDY_SUMMARY respectively6, 36—relative to DMSO controls. No concomitant reduction in the ST:STUDY_SUMMARY levels of the respective phosphorylated products, PI(3,5)P2 and PI(4,5)P2, was ST:STUDY_SUMMARY detected in RMC-113- vs. DMSO-treated infected cells, likely due to intact ST:STUDY_SUMMARY activity of enzymes not targeted by RMC-113, such as PIP4K (1A/1B/1C). ST:STUDY_SUMMARY Nevertheless, the product-to-substrate ratios of PIKfyve and PIP4K2C were ST:STUDY_SUMMARY reduced in both uninfected and infected cells upon RMC-113 treatment relative to ST:STUDY_SUMMARY DMSO ST:INSTITUTE Stanford University ST:DEPARTMENT medicine ST:LABORATORY Einav lab ST:LAST_NAME Mishra ST:FIRST_NAME Manjari ST:ADDRESS 300 pasteur drive, stanford ST:EMAIL manjari1@stanford.edu ST:PHONE 6503849709 ST:NUM_GROUPS 4 (uninfected and SARS-CoV-2 infected samples and RMC-113-treated and untreated ST:NUM_GROUPS samples) ST:STUDY_COMMENTS na ST:PUBLICATIONS https://www.biorxiv.org/content/10.1101/2024.04.15.589676v1.full #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:CELL_BIOSOURCE_OR_SUPPLIER ATCC SU:CELL_STRAIN_DETAILS Human lung epithelial cells (A549-ACE2) SU:CELL_PASSAGE_NUMBER 15 SU:CELL_COUNTS 1*10^6 cells #FACTORS #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS 1 DMSO SARS-CoV2 Sample source:A549-ACE2 CELLS | Treatment:Control RAW_FILE_NAME(raw file)=PI.WIFF, PIP.WIFF,PIP2.WIFF,PIP3.WIFF SUBJECT_SAMPLE_FACTORS 2 RMC-113 SARS-CoV2 Sample source:A549-ACE2 cells | Treatment:treated RAW_FILE_NAME(raw file)=PI.WIFF, PIP.WIFF,PIP2.WIFF,PIP3.WIFF SUBJECT_SAMPLE_FACTORS 3 DMSO uninfected Sample source:A549ACE2 CELLS | Treatment:Control RAW_FILE_NAME(raw file)=PI.WIFF, PIP.WIFF,PIP2.WIFF,PIP3.WIFF SUBJECT_SAMPLE_FACTORS 4 RMC-113 uninfected Sample source:A549ACE2 CELLS | Treatment:treated RAW_FILE_NAME(raw file)=PI.WIFF, PIP.WIFF,PIP2.WIFF,PIP3.WIFF #COLLECTION CO:COLLECTION_SUMMARY In this study, we have used lung epithelial A549-ACE2 cell line for our CO:COLLECTION_SUMMARY experiment in both SARS-CoV2 infected and uninfected cells including treatment CO:COLLECTION_SUMMARY with our lab-made dual lipid kinase inhibitor RMC-113 CO:SAMPLE_TYPE Epithelial cells #TREATMENT TR:TREATMENT_SUMMARY The cells are treated with dual lipid kinase inhibitor RMC-113 TR:TREATMENT_DOSE 5uM TR:TREATMENT_DOSEDURATION 24hrs TR:TREATMENT_VEHICLE methanol TR:CELL_STORAGE 37 Celsius 5% co2 TR:CELL_GROWTH_CONTAINER petriplate TR:CELL_GROWTH_RATE 32hrs doubling time TR:CELL_MEDIA DMEM TR:CELL_ENVIR_COND ambient TR:CELL_HARVESTING p-16 TR:CELL_PCT_CONFLUENCE 80% #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Lipid extraction: 10 pmol each of 4 reference standards (deuterated 5 PI, SP:SAMPLEPREP_SUMMARY deuterated 62 PI3P, deuterated 62 PI(4,5)P2) was spiked into methanol volume SP:SAMPLEPREP_SUMMARY required for total # of samples. 1 sample is spiked with 37:4 and 38:4 reference SP:SAMPLEPREP_SUMMARY standards (Table 1) for retention time matching. 1.5 ml of methanol standard mix SP:SAMPLEPREP_SUMMARY was added to 200 ul of plasma or cell pellet. 750 of uL Ultrapure water, 750ul SP:SAMPLEPREP_SUMMARY of 2M HCL, and 200 uL of 1 M NaCl were added to above mixture. Mixture was SP:SAMPLEPREP_SUMMARY vortexed, then 1 mL of chloroform was added. Tubes were mixed for 2 min. After SP:SAMPLEPREP_SUMMARY centrifugation at 1200×g for 4 min at RT, lower organic was transferred to a SP:SAMPLEPREP_SUMMARY new tube. Prep for Methylation: 1 mL of Methanol/12 M HCL/H20 (12/12/1/1) was SP:SAMPLEPREP_SUMMARY added to the lower phase. 900 uL of 120 mM NaCl was added to the mixture and SP:SAMPLEPREP_SUMMARY vortexed. After centrifugation at 1200×g for 4 min at RT, lower organic was SP:SAMPLEPREP_SUMMARY transferred to a new tube. Methylation reaction: 50 uL of 2 M trimethylsilyl SP:SAMPLEPREP_SUMMARY diazomethane was added to each sample tube. After 10 min, the reaction was SP:SAMPLEPREP_SUMMARY quenched with 20 uL of glacial acetic acid. Samples were mixed with 700 uL of SP:SAMPLEPREP_SUMMARY chloroform/methanol/UltrapureH20 (3/48/47). Tubes were mixed for 1 min. After SP:SAMPLEPREP_SUMMARY centrifugation at 1200×g for 4 min at RT, lower organic was transferred to a SP:SAMPLEPREP_SUMMARY 1.5 mL vials dried under N2 stream. Samples were redissolved in 200 uL SP:SAMPLEPREP_SUMMARY acetonitrile. Samples were placed on shaker for 10 minutes, then transferred to SP:SAMPLEPREP_SUMMARY a 200 ul glass vial insert. Samples were loaded on UPLC autosampler and injected SP:SAMPLEPREP_SUMMARY at 30 uL for LCMS analysis. SP:PROCESSING_STORAGE_CONDITIONS 4℃ SP:EXTRACT_STORAGE 4℃ SP:SAMPLE_DERIVATIZATION methylation SP:SAMPLE_SPIKING yes #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Setup Chiral column to MS and create MRM profiles for each lipid class. Run CH:CHROMATOGRAPHY_SUMMARY samples and collect data using parameters in table 2 and 3 below. Table 2. UHPLC CH:CHROMATOGRAPHY_SUMMARY Parameters Column: Lux 3 µm i-Cellulose-5, LC Column 250 x 4.6 mm, Part No.: CH:CHROMATOGRAPHY_SUMMARY 00G-4755-E0 Detector: MS triple quad Sciex 5500 Injection Volume: 30 µL Sample CH:CHROMATOGRAPHY_SUMMARY Temperature: 15 °C Column Temperature: 35 °C Flow rate: 0.5 mL/min. Mobile CH:CHROMATOGRAPHY_SUMMARY Phase-A: 5 mM ammonium acetate in Methanol Mobile Phase-B: 5 mM ammonium acetate CH:CHROMATOGRAPHY_SUMMARY in Acetonitrile Elution Mode: Gradient see below table Time (min.) A% B% 0 0 100 CH:CHROMATOGRAPHY_SUMMARY 1 0 100 3 30 70 15 30 70 15.01 0 100 20 0 100 CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Shimadzu Nexera X2 CH:COLUMN_NAME Phenomenex Lux i-Cellulose-5 (250 x 4.6mm, 3um) CH:SOLVENT_A 100% methanol; 5 mM ammonium acetate CH:SOLVENT_B 100% acetonitrile; 5 mM ammonium acetate CH:SOLVENT_C none CH:FLOW_GRADIENT Time (min.) A% B%; 0 0 100; 1 0 100; 3 30 70; 15 30 70; 15.01 0 100; 20 0 100 CH:FLOW_RATE 0.5 mL/min CH:COLUMN_TEMPERATURE 35 CH:METHODS_FILENAME LIPIDOMICS_protocol_all_parameters_etc.pdf CH:INTERNAL_STANDARD PI, PIP, PIP2 #ANALYSIS AN:ANALYSIS_TYPE MS AN:LABORATORY_NAME Einav lab AN:OPERATOR_NAME Adam AN:SOFTWARE_VERSION analyst 3.0 #MS MS:INSTRUMENT_NAME ABI Sciex 5500 QTrap MS:INSTRUMENT_TYPE Other MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Calculate areas under curve of each sample with retention times matching the MS:MS_COMMENTS 37:4 and 38:4 standard mix only sample. Divide areas of each sample by their MS:MS_COMMENTS respective deuterated reference standard to account for experimental and MS:MS_COMMENTS analytical variation. Use analyst 3.0 software to both run samples and analyze MS:MS_COMMENTS and process samples #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS au MS_METABOLITE_DATA_START Samples DMSO SARS-CoV2 RMC-113 SARS-CoV2 DMSO uninfected RMC-113 uninfected Factors Sample source:A549-ACE2 CELLS | Treatment:Control Sample source:A549-ACE2 cells | Treatment:treated Sample source:A549ACE2 CELLS | Treatment:Control Sample source:A549ACE2 CELLS | Treatment:treated PI32:1 9.51E+06 6.92E+06 9.65E+06 6.54E+06 PI32:0 PI34:2 3.51E+07 2.77E+07 3.45E+07 2.68E+07 PI34:1 4.14E+07 2.82E+07 4.11E+07 2.65E+07 PI34:0 3.87E+06 2.52E+06 3.87E+06 2.41E+06 PI36:4 9.06E+06 5.87E+06 8.12E+06 6.21E+06 PI36:3 1.39E+07 1.02E+07 1.25E+07 1.05E+07 PI36:2 7.53E+07 4.75E+07 7.07E+07 4.71E+07 PI36:1 5.62E+07 3.43E+07 5.22E+07 3.29E+07 PI36:0 5.59E+06 3.20E+06 5.22E+06 3.07E+06 PI37:4 1.41E+06 1.17E+06 1.30E+06 1.26E+06 PI38:6 3.09E+06 1.64E+06 2.33E+06 1.66E+06 PI38:5 3.27E+07 2.20E+07 2.83E+07 2.38E+07 PI38:4 7.65E+07 6.43E+07 7.47E+07 6.77E+07 PI38:3 6.09E+07 4.15E+07 5.62E+07 3.96E+07 PI40:6 8.16E+06 5.72E+06 7.21E+06 5.95E+06 PI40:5 9.66E+06 8.89E+06 9.57E+06 9.62E+06 PI40:4 6.39E+06 5.32E+06 5.82E+06 5.62E+06 PI3P32:1 1.36E+05 8.27E+04 1.01E+05 9.38E+04 PI3P32:0 PI3P34:2 2.22E+05 2.28E+05 1.78E+05 2.60E+05 PI3P34:1 8.53E+05 5.70E+05 5.90E+05 6.71E+05 PI3P34:0 6.40E+04 3.81E+04 4.12E+04 5.22E+04 PI3P36:4 5.60E+04 4.54E+04 2.84E+04 5.33E+04 PI3P36:3 1.27E+05 1.24E+05 8.66E+04 1.30E+05 PI3P36:2 8.27E+05 6.90E+05 6.05E+05 8.71E+05 PI3P36:1 1.32E+06 9.22E+05 9.29E+05 1.18E+06 PI3P36:0 8.31E+04 5.75E+04 5.49E+04 7.98E+04 PI3P37:4 5.14E+03 7.49E+03 4.26E+03 5.81E+03 PI3P38:6 9.28E+03 9.54E+03 5.32E+03 9.28E+03 PI3P38:5 1.71E+05 1.72E+05 8.84E+04 2.00E+05 PI3P38:4 7.61E+05 9.74E+05 5.27E+05 1.23E+06 PI3P38:3 9.26E+05 9.30E+05 5.81E+05 1.17E+06 PI3P40:6 6.07E+04 4.85E+04 4.19E+04 7.59E+04 PI3P40:5 9.59E+04 1.26E+05 5.83E+04 1.38E+05 PI3P40:4 1.07E+05 1.31E+05 6.82E+04 1.49E+05 PI4P32:1 5.38E+05 3.80E+05 3.72E+05 3.43E+05 PI4P32:0 PI4P34:2 1.98E+06 1.52E+06 1.17E+06 1.42E+06 PI4P34:1 3.48E+06 2.20E+06 2.35E+06 2.01E+06 PI4P34:0 3.11E+05 2.01E+05 2.07E+05 2.01E+05 PI4P36:4 4.50E+05 3.15E+05 2.79E+05 3.02E+05 PI4P36:3 8.07E+05 5.80E+05 4.13E+05 5.60E+05 PI4P36:2 6.16E+06 3.91E+06 3.71E+06 3.93E+06 PI4P36:1 4.10E+06 2.79E+06 2.73E+06 2.70E+06 PI4P36:0 3.57E+05 2.49E+05 2.46E+05 2.44E+05 PI4P37:4 3.26E+05 2.82E+05 2.94E+05 2.98E+05 PI4P38:6 8.09E+04 6.86E+04 5.02E+04 5.96E+04 PI4P38:5 1.36E+06 1.10E+06 7.76E+05 1.15E+06 PI4P38:4 5.40E+06 4.94E+06 3.08E+06 5.21E+06 PI4P38:3 3.84E+06 3.07E+06 2.30E+06 3.05E+06 PI4P40:6 3.94E+05 3.34E+05 2.46E+05 3.36E+05 PI4P40:5 5.24E+05 6.08E+05 3.45E+05 6.25E+05 PI4P40:4 3.79E+05 4.20E+05 2.27E+05 4.72E+05 PI5P32:1 4.08E+03 1.65E+03 3.20E+03 2.83E+03 PI5P32:0 PI5P34:2 2.64E+04 2.20E+04 2.31E+04 2.26E+04 PI5P34:1 4.78E+04 4.68E+04 4.58E+04 4.69E+04 PI5P34:0 5.89E+04 3.33E+04 4.10E+04 3.05E+04 PI5P36:4 5.06E+02 1.60E+03 1.36E+03 7.99E+02 PI5P36:3 2.97E+03 4.02E+03 3.62E+03 6.18E+03 PI5P36:2 6.93E+04 5.90E+04 5.38E+04 6.10E+04 PI5P36:1 4.70E+04 6.87E+04 4.76E+04 8.36E+04 PI5P36:0 9.17E+03 8.89E+03 8.11E+03 1.07E+04 PI5P37:4 2.92E+04 4.68E+04 1.99E+04 4.96E+04 PI5P38:6 PI5P38:5 8.17E+02 3.06E+03 1.15E+03 3.11E+03 PI5P38:4 6.09E+04 1.05E+05 5.10E+04 1.19E+05 PI5P38:3 1.10E+04 2.40E+04 1.43E+03 1.32E+04 PI5P40:6 9.60E+02 2.59E+03 4.90E+03 5.02E+03 PI5P40:5 7.80E+03 1.48E+04 4.12E+03 1.54E+04 PI5P40:4 3.17E+03 6.95E+03 4.81E+03 7.28E+03 PI(3,5)P232:1 6.86E+03 3.84E+03 3.62E+03 2.25E+03 PI(3,5)P232:0 PI(3,5)P234:2 2.38E+04 1.66E+04 1.68E+04 1.26E+04 PI(3,5)P234:1 3.56E+04 1.52E+04 1.67E+04 1.44E+04 PI(3,5)P234:0 PI(3,5)P236:4 9.59E+03 7.50E+03 5.07E+03 6.41E+03 PI(3,5)P236:3 1.08E+04 6.82E+03 5.60E+03 6.05E+03 PI(3,5)P236:2 6.61E+04 3.21E+04 3.56E+04 3.11E+04 PI(3,5)P236:1 5.17E+04 2.72E+04 2.84E+04 2.94E+04 PI(3,5)P236:0 5.91E+03 3.37E+03 3.60E+03 3.06E+03 PI(3,5)P237:4 7.15E+03 5.78E+03 6.24E+03 4.98E+03 PI(3,5)P238:6 PI(3,5)P238:5 2.70E+04 2.23E+04 1.43E+04 2.28E+04 PI(3,5)P238:4 7.30E+04 6.85E+04 4.49E+04 6.48E+04 PI(3,5)P238:3 5.20E+04 3.52E+04 2.80E+04 3.44E+04 PI(3,5)P240:6 6.43E+03 5.31E+03 3.85E+03 3.59E+03 PI(3,5)P240:5 7.43E+03 0.00E+00 4.65E+03 8.92E+03 PI(3,5)P240:4 5.12E+03 4.72E+03 4.33E+03 4.64E+03 PI(4,5)P2 32:1 5.17E+05 4.17E+05 3.61E+05 3.43E+05 PI(4,5)P232:0 PI(4,5)P234:2 1.58E+06 1.38E+06 9.61E+05 1.18E+06 PI(4,5)P234:1 2.88E+06 1.99E+06 1.77E+06 1.80E+06 PI(4,5)P234:0 PI(4,5)P236:4 2.43E+05 2.02E+05 1.47E+05 2.10E+05 PI(4,5)P236:3 5.62E+05 5.03E+05 3.26E+05 4.96E+05 PI(4,5)P236:2 4.15E+06 2.95E+06 2.49E+06 2.94E+06 PI(4,5)P236:1 4.45E+06 3.36E+06 3.06E+06 3.36E+06 PI(4,5)P236:0 3.93E+05 2.97E+05 2.66E+05 2.86E+05 PI(4,5)P237:4 2.30E+05 2.27E+05 2.27E+05 2.16E+05 PI(4,5)P238:6 PI(4,5)P238:5 6.98E+05 6.90E+05 4.05E+05 7.31E+05 PI(4,5)P238:4 3.52E+06 3.53E+06 2.14E+06 3.77E+06 PI(4,5)P238:3 2.67E+06 2.20E+06 1.51E+06 2.25E+06 PI(4,5)P240:6 2.07E+05 2.07E+05 1.34E+05 2.29E+05 PI(4,5)P240:5 3.73E+05 4.31E+05 2.43E+05 4.91E+05 PI(4,5)P240:4 2.37E+05 2.73E+05 1.48E+05 2.96E+05 PI(3,4,5)P3 32:1 PI(3,4,5)P3 32:0 2.34E+05 2.50E+05 2.19E+05 1.80E+05 PI(3,4,5)P334:2 PI(3,4,5)P334:1 PI(3,4,5)P334:0 9.56E+04 1.03E+05 9.32E+04 7.75E+04 PI(3,4,5)P336:4 1.92E+04 1.05E+04 1.41E+04 4.77E+04 PI(3,4,5)P336:3 PI(3,4,5)P336:2 PI(3,4,5)P336:1 PI(3,4,5)P336:0 PI(3,4,5)P337:4 7.21E+03 6.07E+03 3.15E+02 6.41E+03 PI(3,4,5)P338:6 PI(3,4,5)P338:5 9.19E+04 6.77E+04 7.03E+04 4.28E+04 PI(3,4,5)P338:4 1.34E+04 1.24E+04 9.04E+03 6.86E+03 PI(3,4,5)P338:3 PI(3,4,5)P340:6 PI(3,4,5)P340:5 PI(3,4,5)P340:4 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name PI32:1 PI32:0 PI34:2 PI34:1 PI34:0 PI36:4 PI36:3 PI36:2 PI36:1 PI36:0 PI37:4 PI38:6 PI38:5 PI38:4 PI38:3 PI40:6 PI40:5 PI40:4 PI3P32:1 PI3P32:0 PI3P34:2 PI3P34:1 PI3P34:0 PI3P36:4 PI3P36:3 PI3P36:2 PI3P36:1 PI3P36:0 PI3P37:4 PI3P38:6 PI3P38:5 PI3P38:4 PI3P38:3 PI3P40:6 PI3P40:5 PI3P40:4 PI4P32:1 PI4P32:0 PI4P34:2 PI4P34:1 PI4P34:0 PI4P36:4 PI4P36:3 PI4P36:2 PI4P36:1 PI4P36:0 PI4P37:4 PI4P38:6 PI4P38:5 PI4P38:4 PI4P38:3 PI4P40:6 PI4P40:5 PI4P40:4 PI5P32:1 PI5P32:0 PI5P34:2 PI5P34:1 PI5P34:0 PI5P36:4 PI5P36:3 PI5P36:2 PI5P36:1 PI5P36:0 PI5P37:4 PI5P38:6 PI5P38:5 PI5P38:4 PI5P38:3 PI5P40:6 PI5P40:5 PI5P40:4 PI(3,5)P232:1 PI(3,5)P232:0 PI(3,5)P234:2 PI(3,5)P234:1 PI(3,5)P234:0 PI(3,5)P236:4 PI(3,5)P236:3 PI(3,5)P236:2 PI(3,5)P236:1 PI(3,5)P236:0 PI(3,5)P237:4 PI(3,5)P238:6 PI(3,5)P238:5 PI(3,5)P238:4 PI(3,5)P238:3 PI(3,5)P240:6 PI(3,5)P240:5 PI(3,5)P240:4 PI(4,5)P2 32:1 PI(4,5)P232:0 PI(4,5)P234:2 PI(4,5)P234:1 PI(4,5)P234:0 PI(4,5)P236:4 PI(4,5)P236:3 PI(4,5)P236:2 PI(4,5)P236:1 PI(4,5)P236:0 PI(4,5)P237:4 PI(4,5)P238:6 PI(4,5)P238:5 PI(4,5)P238:4 PI(4,5)P238:3 PI(4,5)P240:6 PI(4,5)P240:5 PI(4,5)P240:4 PI(3,4,5)P3 32:1 PI(3,4,5)P3 32:0 PI(3,4,5)P334:2 PI(3,4,5)P334:1 PI(3,4,5)P334:0 PI(3,4,5)P336:4 PI(3,4,5)P336:3 PI(3,4,5)P336:2 PI(3,4,5)P336:1 PI(3,4,5)P336:0 PI(3,4,5)P337:4 PI(3,4,5)P338:6 PI(3,4,5)P338:5 PI(3,4,5)P338:4 PI(3,4,5)P338:3 PI(3,4,5)P340:6 PI(3,4,5)P340:5 PI(3,4,5)P340:4 METABOLITES_END #END