#METABOLOMICS WORKBENCH Jung_DNLI_20250214_171815 DATATRACK_ID:5629 STUDY_ID:ST003737 ANALYSIS_ID:AN006135 PROJECT_ID:PR002323 VERSION 1 CREATED_ON February 16, 2025, 11:41 am #PROJECT PR:PROJECT_TITLE PILRA regulates microglial neuroinflammation and lipid metabolism as a candidate PR:PROJECT_TITLE therapeutic target for Alzheimer’s disease PR:PROJECT_TYPE Cellular studies PR:PROJECT_SUMMARY The Alzheimer’s disease (AD) human genetic landscape identified microglia as a PR:PROJECT_SUMMARY key disease-modifying brain cell type. A common loss-of-function coding variant PR:PROJECT_SUMMARY in paired immunoglobulin-like type 2 receptor alpha (PILRA) associated with PR:PROJECT_SUMMARY reduced AD risk, was found enriched in a cohort of healthy centenarians and PR:PROJECT_SUMMARY rescued APOE4 AD risk, however the mechanisms underlying protection are PR:PROJECT_SUMMARY undefined. Here we identify biological functions of PILRA, an immunoreceptor PR:PROJECT_SUMMARY tyrosine-based inhibitory motif (ITIM)-containing receptor, in human PR:PROJECT_SUMMARY iPSC-derived microglia (iMG) and chimeric AD mice. CRISPR-mediated PILRA PR:PROJECT_SUMMARY knockout (KO) in iMG rescued ApoE4-induced immunometabolic deficits. Moreover, PR:PROJECT_SUMMARY loss of PILRA confers a signature of metabolic resilience in microglia with PR:PROJECT_SUMMARY increased mitochondrial capacity in tandem with elevated antioxidants, reduced PR:PROJECT_SUMMARY ROS and toxic lipid species. Additionally, PILRA KO iMG exhibit improved PR:PROJECT_SUMMARY lysosomal degradation, enhanced migration, and attenuated cytokine responses. We PR:PROJECT_SUMMARY show PPAR and STAT1/3 act as master regulators that mediate downstream signaling PR:PROJECT_SUMMARY and regulate PILRA-dependent microglial functions. Importantly, AD mice PR:PROJECT_SUMMARY transplanted with human PILRA KO microglia showed reduced amyloid pathology and PR:PROJECT_SUMMARY rescued levels of synaptic markers. Finally, we identify a high-affinity human PR:PROJECT_SUMMARY PILRA-specific antagonist antibody that phenocopies PILRA KO. Together, these PR:PROJECT_SUMMARY findings suggest a therapeutic approach to modulate microglial immunometabolism PR:PROJECT_SUMMARY by inhibiting PILRA, thus identifying a pharmacologically tractable target for PR:PROJECT_SUMMARY AD. PR:INSTITUTE Denali Therapeutics PR:LAST_NAME Suh PR:FIRST_NAME Jung PR:ADDRESS 161 Oyster Point Blvd, South San Francisco, CA 94080 PR:EMAIL suh@dnli.com PR:PHONE +1 6507973837 #STUDY ST:STUDY_TITLE PILRA regulates microglial neuroinflammation and lipid metabolism as a candidate ST:STUDY_TITLE therapeutic target for Alzheimer’s disease ST:STUDY_TYPE In vitro study ST:STUDY_SUMMARY The Alzheimer’s disease (AD) human genetic landscape identified microglia as a ST:STUDY_SUMMARY key disease-modifying brain cell type. A common loss-of-function coding variant ST:STUDY_SUMMARY in paired immunoglobulin-like type 2 receptor alpha (PILRA) associated with ST:STUDY_SUMMARY reduced AD risk, was found enriched in a cohort of healthy centenarians and ST:STUDY_SUMMARY rescued APOE4 AD risk, however the mechanisms underlying protection are ST:STUDY_SUMMARY undefined. Here we identify biological functions of PILRA, an immunoreceptor ST:STUDY_SUMMARY tyrosine-based inhibitory motif (ITIM)-containing receptor, in human ST:STUDY_SUMMARY iPSC-derived microglia (iMG) and chimeric AD mice. CRISPR-mediated PILRA ST:STUDY_SUMMARY knockout (KO) in iMG rescued ApoE4-induced immunometabolic deficits. Moreover, ST:STUDY_SUMMARY loss of PILRA confers a signature of metabolic resilience in microglia with ST:STUDY_SUMMARY increased mitochondrial capacity in tandem with elevated antioxidants, reduced ST:STUDY_SUMMARY ROS and toxic lipid species. Additionally, PILRA KO iMG exhibit improved ST:STUDY_SUMMARY lysosomal degradation, enhanced migration, and attenuated cytokine responses. We ST:STUDY_SUMMARY show PPAR and STAT1/3 act as master regulators that mediate downstream signaling ST:STUDY_SUMMARY and regulate PILRA-dependent microglial functions. Importantly, AD mice ST:STUDY_SUMMARY transplanted with human PILRA KO microglia showed reduced amyloid pathology and ST:STUDY_SUMMARY rescued levels of synaptic markers. Finally, we identify a high-affinity human ST:STUDY_SUMMARY PILRA-specific antagonist antibody that phenocopies PILRA KO. Together, these ST:STUDY_SUMMARY findings suggest a therapeutic approach to modulate microglial immunometabolism ST:STUDY_SUMMARY by inhibiting PILRA, thus identifying a pharmacologically tractable target for ST:STUDY_SUMMARY AD. ST:INSTITUTE Denali Therapeutics ST:LAST_NAME Suh ST:FIRST_NAME Jung ST:ADDRESS 161 Oyster Point Blvd, South San Francisco, CA 94080 ST:EMAIL suh@dnli.com ST:PHONE +1 06507973837 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:GENOTYPE_STRAIN WT, PILRA KO SU:CELL_BIOSOURCE_OR_SUPPLIER University of California Irvine SU:CELL_STRAIN_DETAILS Induced pluripotent stem cells derived microglia cells SU:CELL_PRIMARY_IMMORTALIZED no SU:CELL_COUNTS 500,000 SU:SPECIES_GROUP human #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 - HSA-000033340 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_002.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_002.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_002.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_002.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033323 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_003.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_003.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_003.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_003.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033319 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_004.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_004.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_004.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_004.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033335 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_005.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_005.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_005.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_005.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033332 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_006.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_006.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_006.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_006.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033327 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_007.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_007.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_007.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_007.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033331 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_008.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_008.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_008.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_008.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033329 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_009.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_009.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_009.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_009.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033326 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_010.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_010.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_010.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_010.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033334 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_011.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_011.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_011.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_011.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033324 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_012.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_012.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_012.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_012.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033322 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_013.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_013.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_013.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_013.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033320 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_015.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_015.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_015.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_015.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033337 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_016.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_016.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_016.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_016.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033317 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_017.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_017.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_017.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_017.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033339 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_018.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_018.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_018.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_018.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033330 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_019.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_019.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_019.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_019.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033328 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_020.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_020.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_020.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_020.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033325 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_021.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_021.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_021.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_021.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033321 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_022.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_022.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_022.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_022.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033336 genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_023.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_023.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_023.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_023.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033338 genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_024.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_024.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_024.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_024.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033318 genotype:WT | treatment:Vehicle | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_025.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_025.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_025.mzML; cell_line=Line 1 DNLI; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_025.mzML SUBJECT_SAMPLE_FACTORS - HSA-000033333 genotype:WT | treatment:LPS | Sample source:iPSC microglia RAW_FILE_NAME(Met_Neg)=20230911_TW_HEX-000666_Met_Neg_026.mzML; RAW_FILE_NAME(Lipid_Pos)=20230908_TW_HEX-000666_Lipid_Pos_026.mzML; RAW_FILE_NAME(Lipid_Neg)=20230909_TW_HEX-000666_Lipid_Neg_026.mzML; cell_line=Line 2 MBJ; RAW_FILE_NAME(Met_Pos)=20230909_TW_HEX-000666_Met_Pos_026.mzML #COLLECTION CO:COLLECTION_SUMMARY WT and PILRA KO microglia cells (500,000) were pelleted, and washed 1x with 0.9% CO:COLLECTION_SUMMARY sodium chloride and collected in collected in Lobind 1.5 mL Eppendorf tubes and CO:COLLECTION_SUMMARY snap frozen. CO:SAMPLE_TYPE iPSC cells CO:STORAGE_CONDITIONS -80℃ CO:COLLECTION_VIALS Lobind 1.5 mL Eppendorf tubes CO:STORAGE_VIALS Lobind 1.5 mL Eppendorf tubes #TREATMENT TR:TREATMENT_SUMMARY WT and PILRA KO microglia cells were cultured in iPSC-complete media without TR:TREATMENT_SUMMARY TGFBeta and plated on matrigel-coated 12-well plate. Wells were supplemented TR:TREATMENT_SUMMARY with PBS (vehicle) or endotoxin (LPS) (100ng/ml, Sigma-Aldrich, 055-B5) for 48 TR:TREATMENT_SUMMARY hours and harvested for metabolite extraction. TR:TREATMENT_COMPOUND endotoxin (LPS; Sigma-Aldrich, 055-B5) TR:TREATMENT_DOSE 100 ng/mL TR:TREATMENT_DOSEDURATION 48 hours TR:TREATMENT_VEHICLE PBS #SAMPLEPREP SP:SAMPLEPREP_SUMMARY On the day of lipid and metabolite extraction, cell pellets were reconstituted SP:SAMPLEPREP_SUMMARY with 400 μL MS grade methanol spiked with internal standards.Samples were SP:SAMPLEPREP_SUMMARY vortexed and volumes were adjusted to 800 μL with MS grade H2O, vortexed again, SP:SAMPLEPREP_SUMMARY then 600 μL tert-butyl methyl ether (MTBE) was added, vortexed, then centrifuged SP:SAMPLEPREP_SUMMARY at 21,000 g for 10 min at 4C. The two phases generated by centrifugation were SP:SAMPLEPREP_SUMMARY separated. Each phase (top: non-polar lipids, bottom: polar metabolites) was SP:SAMPLEPREP_SUMMARY transferred to glass vials and dried overnight using a Genevac EZ3. Non-polar SP:SAMPLEPREP_SUMMARY lipids were resuspended MS grade methanol and polar metabolites were resuspended SP:SAMPLEPREP_SUMMARY in 90% methanol:water mixture. SP:PROCESSING_STORAGE_CONDITIONS On ice SP:EXTRACT_STORAGE -20℃ #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Agilent 1290 Infinity II CH:COLUMN_NAME Waters ACQUITY UPLC BEH Amide (100 x 2.1 mm, 1.7 μm) CH:SOLVENT_A 60% acetonitrile/40% water; 10 mM ammonium acetate; 0.1% acetic acid CH:SOLVENT_B 90% isopropyl alcohol/10% acetonitrile; 10 mM ammonium acetate; 0.1% acetic acid CH:FLOW_GRADIENT 0.0-8.0 min from 45% B to 99% B, 8.0-9.0 min at 99% B, 9.0-9.1 min to 45% B, and CH:FLOW_GRADIENT 9.1-10.0 min at 45% B CH:FLOW_RATE 0.25 mL/min CH:COLUMN_TEMPERATURE 55°C #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME ABI Sciex 6500+ QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Curtain gas at 40 psi (positive mode); collision gas was set at medium; ion MS:MS_COMMENTS spray voltage at -4500 V ; temperature at 250°C ; ion source Gas 1 at 55 psi; MS:MS_COMMENTS ion source Gas 2 at 60 psi; entrance potential at - 10 V; and collision cell MS:MS_COMMENTS exit potential at -15 V. Quantification was performed using MultiQuant 3.02 MS:MS_COMMENTS (Sciex). #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS area MS_METABOLITE_DATA_START Samples HSA-000033340 HSA-000033323 HSA-000033319 HSA-000033335 HSA-000033332 HSA-000033327 HSA-000033331 HSA-000033329 HSA-000033326 HSA-000033334 HSA-000033324 HSA-000033322 HSA-000033320 HSA-000033337 HSA-000033317 HSA-000033339 HSA-000033330 HSA-000033328 HSA-000033325 HSA-000033321 HSA-000033336 HSA-000033338 HSA-000033318 HSA-000033333 Factors genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia genotype:PILRAKO | treatment:Vehicle | Sample source:iPSC microglia genotype:PILRAKO | treatment:LPS | Sample source:iPSC microglia genotype:WT | treatment:Vehicle | Sample source:iPSC microglia genotype:WT | treatment:LPS | Sample source:iPSC microglia (3-O-sulfo)GalCer(d18:1/16:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/18:0(2OH) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/18:0(d3)) 11264496.22 12148421.17 12962654.3 14331345.18 11010343.53 10879391.95 11609832.69 13354034.35 13698707.26 12085037.88 11833038.53 14055807.95 13930172.73 10319757.91 10678244.28 11177319.5 12471623.82 12164800.43 12398983.6 13319692.44 10322595.92 12196777.56 13686195.6 13294357.93 (3-O-sulfo)GalCer(d18:1/18:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/24:0(2OH)) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/24:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/24:1(2OH)) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA (3-O-sulfo)GalCer(d18:1/24:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Arachidonic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Arachidonic acid_MRM NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Arachidonic acid-d8 52489628.88 45994651.53 50464171.04 55674104.28 52632456.86 54321602.48 46558710.11 52984358.41 55769466.96 43836741.78 44871139.27 47104484.51 53186252.84 37768639.66 61368511.66 42285469.9 46568270.15 44938154.14 50093641.92 48889560.13 47901063.05 47645563.3 51876081.7 52505135.38 Arachidonic acid-d8_MRM 25094627.04 25280731.09 24627296.88 24142814.08 24153253.91 24439996.77 22827961.5 23414791.29 25996862.68 22624119.83 23010840.91 24934040.34 24344053.22 21030847.46 28735929.48 23662075.93 22582784.25 22254816.04 21908329.55 24199688.18 22775469.35 21688451.09 24129273.63 24273582.05 BMP(14:0/14:0) 53465230.04 51998736.46 56168387.31 54411774.19 52322591.65 55228624.63 46418955.49 54298083.9 53953221.01 53559660.9 50502584.39 52236172.83 54899608.79 44373867.06 51431267.82 54383494.61 51903866.5 48931918.75 52345815.98 48855094.15 52369609.09 47839241.6 51192084.64 54070903.79 BMP(16:0_18:1) 15953331.59 30101548.8 45700122.4 11978428.6 12173130.3 34539788.99 6137248.16 10984795.13 35958105.92 4167531.725 23313290.65 55541782.94 63689170.02 8415548.208 45896253.89 12842021.92 7229357.526 26067449.32 19384711.07 55483150.35 11620896.04 6867166.733 47967678.99 12637798.23 BMP(16:0_20:4) 4283662.097 6236575.497 11028534.51 3564150.423 4007909.818 6813251.209 2835565.674 3202435.881 8679756.853 1581817.998 6969311.59 11639392.03 14463343.45 3606424.516 11575895.51 3259901.078 2509165.668 4575026.383 3882642.854 13428639.58 3652447.159 1933697.724 12380400.44 2966257.106 BMP(16:0_22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA BMP(16:1/16:1) 1132025.846 2303261.688 4973471.384 926519.2158 738978.0075 2748341.283 500143.2111 641867.0422 3255678.81 409178.4112 1993608.478 6540508.2 5113471.95 823076.0255 5088159.699 1227293.737 685212.7275 1881236.432 1829713.613 6586848.753 1054581.259 738486.512 6040865.973 1033624.78 BMP(18:0_18:1) 8065076.558 20089015.26 36579366.74 9059661.268 8918051.053 26346446.23 6166071.957 7789647.488 27157965.18 4528117.757 16876786.98 43957920.19 48932487.32 6687082.098 35692227.25 7884922.552 7608421.398 13490924.45 12646327.46 45670513.17 6905140.227 3821190.068 36922786.37 9389521.059 BMP(18:0_20:4) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA BMP(18:0_22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA BMP(18:1/18:1) 42171353.81 77528163.7 99749468.2 39532622.58 35020684.74 84275901.81 24567417.13 36200887.89 87174196.41 20682312.94 71403133.5 100993434.4 109425882 29382677.35 99044858.31 39268184.37 25921796.27 66754202.12 64257127.79 107753763.4 40167878.64 23900491.25 94985143.66 40017891.68 BMP(20:4/20:4) 3272208.334 4345298.554 6413988.577 3791898.682 4295605.029 4527461.145 3845668.564 4173238.336 4527977.069 3454505.03 5670858.077 3133313.798 2557300.87 2899442.257 3419562.604 4657607.737 4838084.692 4091678.962 4412218.844 2577792.119 4972389.423 2674165.093 4939963.287 4397724.341 BMP(22:6/22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Cholesterol sulfate 4266334.823 7092031.844 1179894.271 1260430.105 905383.3363 776434.8142 686989.1436 3406520.72 1256903.151 1699395.198 2774524.278 574887.5904 1713696.075 2077261.848 1034626.95 1796523.84 740946.2883 1935156.615 1484735.643 1065656.99 2240213.721 1657239.592 875930.9153 602109.2873 CL(14:0/14:0/14:0/14:0) 16530553.01 15285579.31 15453260.34 17336812.3 15421218.08 15369045.78 14017071.77 13923872.43 17082796.65 18144577.43 15954168.93 14731486.31 16294788.18 13324238 13444994.09 16213633.19 15565363.15 15031665.8 16270851.78 15023334.39 15685213.67 16792595.76 13619675.09 15995984.56 CL(72:6/18:2) 13399933.7 16951353.77 23334194.96 14661461.73 14614117.73 20337681.84 12340470.2 14114271.98 18921127.18 9283705.028 17157579.61 22337286.76 24661542.46 12561616.42 19456602 12163915.85 9525753.506 15147340.27 16304129.64 22263005.96 13804739.79 12621892.52 25478609.61 14926280.77 CL(72:7/18:2) 33242660.01 34904225.81 39086642.18 31025841.35 31359171.02 38701554.02 29740755.08 30233277.85 36488952.95 28100809.41 33603243.7 38139157.6 41355659.17 31551426.48 37783140.25 29672504.02 26282529.04 29607133.38 30857883.62 35070765.33 31806458.1 28361947.2 41851079.9 34425430.52 CL(72:8-2(OOH)/18:2) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA CL(72:8/18:2) 59318727.58 65216433.11 71057151.64 61045359.14 61580940.77 67896495.12 56269278.76 57109334.2 67607080.26 50103064.65 61216191.12 65659184.1 70817826.58 53858993.55 65231917.66 55627417.85 52482786.98 59593987.49 57031644.23 66397280.29 56788947.71 52356505.1 72276887.44 62261558.27 CL(74:9/18:2) 27601044.88 31313449.99 35995166.72 27953228.64 27145248.5 36761915.58 24924775.49 25732428.38 35604246.38 23662693.62 30728263.19 35869273.06 38278953.08 26113674.82 35933976.15 28914113.49 21186663.79 29565849.75 28729030.9 34862067.52 26320332.13 24118894.05 38299531.97 26771942.78 DHA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA EPA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA FAHFA(18:1/9-O-18:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA GD1a/b(d36:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA GD3(d34:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA GD3(d36:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA GM3(d18:1/18:0(d5)) 6244379.082 5836038.051 5511645.611 5997449.692 5249109.559 5568612.477 5307270.798 7347368.028 6441106.409 5669594.64 5748922.688 4658634.354 4574665.927 4233898.191 6846885.512 6741874.901 6425877.419 5241780.048 6424483.637 5576365.927 6240594.847 5170394.559 5425087.243 6744653.61 GM3(d34:1) 7985202.626 8251003.338 6663576.941 5519658.703 8260063.478 12476961.36 3987806.548 5750669.742 13914052.72 5198610.028 9387244.338 10514914.2 11563215.83 4689217.556 7754920.399 7998322.294 4603715.256 9169315.854 8068864.469 11252214.14 7010125.226 6472704.693 7628149.173 7736257.294 GM3(d36:1) 1706571.354 1115436.961 1106753.507 1842977.105 1275394.558 1816787.717 1126841.81 1347373.658 1742998.206 989903.4457 1314381.014 1521727.063 1900883.488 647675.1466 2017246.675 1811380.162 1244704.405 1404995.82 991206.2066 1723667.788 1810648.789 1297871.419 1109158.296 1632312.304 GQ1b(d36:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(14:0/14:0)_14:0 48919660.43 46858932.81 49136853.31 46710588.72 45789211.97 51130821.35 42745469.58 45106436 47054483.88 47173611.62 44067820.59 46014739.14 49635093.03 40812387.68 48177582.27 47806259.5 46136611.68 44689046.08 46680146.09 45312485.67 45093873.07 43223187.98 49476386.81 45041182.47 Hemi-BMP(18:1/18:1)_16:0 266344.3651 226570.4038 809613.4846 131158.525 222524.7881 379132.3035 63057.53609 100750.0282 430914.4944 218588.2328 552272.705 1518000.189 2790666.732 54857.81304 822409.5882 193372.1289 242524.5842 613599.4924 315023.0833 2978463.866 136052.1482 144547.2157 1274351.485 191410.9882 Hemi-BMP(18:1/18:1)_18:0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(18:1/18:1)_18:1 228982.1235 302714.894 892574.4952 140671.0874 NA 447184.1458 59087.85163 NA 574419.1184 26234.64782 358082.9288 1139253.352 3119761.147 NA 739145.0647 161386.2906 NA 355232.3032 NA 2136258.743 184205.5539 NA 1075812.798 369404.4025 Hemi-BMP(20:4/20:4)_16:0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(20:4/20:4)_18:0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(20:4/20:4)_18:1 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(20:4/20:4)_20:4 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(22:6/22:6)_16:0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(22:6/22:6)_18:0 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(22:6/22:6)_18:1 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Hemi-BMP(22:6/22:6)_22:6 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Linoleic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Linolenic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA LPE(16:0) 17961898.63 11261334.74 13099439.42 9505600.72 11417351.59 11301722.04 9886003.956 12345799.11 14800489.49 8373567.734 13469043.49 15432838.25 15432520.03 9443968.139 19173188.78 16679774.07 10178591 17401680.94 13236360.47 18182283.02 10517250.43 11747608.58 11480311.87 10379680.89 LPE(18:0) 5593985.812 6968462.829 9894578.125 4230410.931 4706341.131 8323791.684 4164022.229 5880771.429 8806987.043 2616623.349 5833030.359 18406482.83 18656549.11 2424723.184 14512047.86 5207407.706 3710448.719 5968059.453 5427647.08 18334246.09 4258187.698 2883548.005 9340309.992 5858796.132 LPE(18:1(d7)) 43162802.6 43440721.9 43268452.8 43736895.86 40497366.76 42383926.58 36362339.25 44506499.49 44518901.32 44154794.88 41389373.52 42157345.23 41084157.31 32712158.27 37629309.95 45050291.15 42261792.16 41898185.09 40033034.52 40398725.58 42089338.85 40433130.48 44431213.96 41903039.7 LPG(16:0) 512420.5522 1358334.324 2271980.961 572901.089 664238.6811 1296156.559 472141.348 695318.2887 886973.7177 NA 1057278.625 2878705.65 4221932.508 425154.2775 1769184.301 471734.4369 492487.2089 708577.3293 865052.5107 2146343.978 NA 647722.6868 2249773.235 591719.6633 LPG(18:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA LPG(18:1) 1136055.55 4987930.891 8653826.889 1175108.253 1544106.529 6209811.833 1078683.039 1926253.996 3780219.392 845257.9469 5336839.554 9562834.802 8952801.842 1121652.276 9208759.11 1295907.233 946300.8626 3369435.347 3542066.135 10199416.06 1312376.718 753501.8231 9560094.385 2174242.027 LPG(20:4) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA LPG(22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA LPI(16:0) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA LPI(18:0) 1557012.586 2168919.824 4496974.422 1550810.756 1988790.231 3685067.71 1873783.637 2664901.499 3664523.185 1087478.664 2208992.716 6697009.596 8831373.522 881263.8626 5420174.956 1735044.71 1345521.596 2541289.757 2613022.531 9164093.383 1191070.564 923217.5989 4003275.548 1662617.072 Oleic acid 39409887.7 31482079.88 48499207.59 25723363.05 21415908.95 28295540.51 40698058.91 39735327.83 37962184.3 42666990.53 45208192.95 22290460.78 23892167.63 39093255.99 38561075.95 46101309.18 39426935.78 31353636.74 29033555.36 45694265.78 27903464.64 46114977.84 30343181.4 24905442.9 PA(15:0/18:1(d7)) 46178902.08 45880540.85 44742916.75 52272431.31 47352077.5 49710648.22 44620638.07 45287338.45 48113041.32 50649673.53 41193529.5 41968999.31 44090203.94 42981176.39 40991797.84 49176938.08 47252885.79 48968161.48 44774514.26 41559954.97 47877423.04 48426446.43 48348826.61 46622299.72 PA(16:0_18:1) 15609987.51 28751852.63 29877714.16 19457907.02 14236046.35 31583047.55 10198641.61 10455095.31 32602992.54 8782673.339 22432904.71 37613936.18 41123042.13 14904819.73 25988478.04 14085384.06 7893761.097 18623009.32 17130511.33 32251849.37 11920058.08 13502290.51 30098919.67 13051825.97 PA(18:0_18:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PA(18:0_20:4) 3844969.495 5412468.373 5397221.091 5417175.905 2051413.2 5731575.893 3656868.159 2197910.289 4943498.664 2223514.164 5091844.824 3493896.043 2953243.035 4365380.204 3382742.657 2952916.603 2110851.797 4642652.398 4936070.81 2790246.336 4060560.14 2342608.457 4508820.293 2706585.727 PA(18:0_22:6) 9527651.822 4917773.049 5727007.207 11187197.43 9031102.25 5299357.627 7740591.227 9828370.326 4184975.804 6347203.956 5130669.4 4674221.666 5281990.161 9628211.901 7265672.76 9723817.906 8025613.558 4736061.268 3475678.536 5630140.184 10250959.56 5947493.648 4888165.811 9580776.162 PA(18:1/18:1) 4928766.548 8227000.431 10708528.57 4634919.962 4147438.408 8519241.018 2831060.77 4506941.334 8725972.502 2058366.886 6147231.62 9209117.104 9850259.028 3256907.853 6696192.549 4086484.722 2119734.555 6520150.272 5688717.889 8320815.613 3152598.198 2554141.382 9119563.655 2657880.804 Palmitic acid 63630728.02 46691179.75 73517589.56 52933829.37 45363932.97 52292563.29 65568736.62 62719288.68 71299431.31 62232299.31 73850290.92 44388688.71 50433546.53 51665552.13 99132988.82 70875446.53 63683658.07 62518113.76 50722541.9 70562894.36 51534109 82355361.17 52179544.41 53483829.23 Palmitoleic acid 16803897.89 20296860.85 11852436.7 18039339 14062583.23 16497504.42 20287590.91 14042614.29 16707369.19 13562970.48 20064644.22 16401250.36 10570312.75 11909114.42 27802836.22 17064107.59 16421800.38 18247106.05 15680950.89 12607003.92 19820983.03 20057033.6 15778488.75 17458246.81 PE(15:0/18:1(d7)) 22298654.72 21048886.25 20920770.45 22225117.91 23583693.26 21635478.33 20744475.5 22796237.33 24322413.01 25440803.73 23893855.38 19597230.2 20672892 18297768.71 20355638.25 22977728.1 21912361.62 21242580.81 20947477.22 19036565.32 21060599.88 21605814.19 20786244.55 22865374.32 PE(O-16:0/20:4) 73868576.73 90458699.63 101501444.3 73741162.74 71191811.22 90828584.79 58260785.5 64338531.92 97019312.64 51183926.56 83452443.16 106982701.2 107202986.4 63309162.09 99330238.4 69370401.82 55579330.62 74592576.06 79813081.87 106035357.2 70190548.38 59952199.18 108519760.8 65372995.59 PE(O-16:0/22:6) 17934325.02 18689164.57 22549311.46 20199090.41 19689386.15 17237593.06 15768850.69 18194882.9 17711157.46 14145414.45 15502212.3 18880045.86 21208657.27 16407959.77 21157446.98 17343151.85 14542965.85 14552661.63 15557295.64 19263217.34 19536665.13 13973347.65 24692828.86 19023037.97 PE(O-18:0/20:4) 32326310.07 48100371.76 58880162.16 33261792.39 29901242.47 53897149.56 23314723.08 28764477.47 49237187.37 21767950.05 46710926.1 60949099.33 68754700.02 30228959.81 58143687.22 33649249.99 20537188.04 41277980.51 37743538.82 59398004.25 35827484.91 25251588.81 59237984.27 31240296.32 PE(O-18:0/22:6) 3306041.482 3806568.66 6140791.945 3009265.623 3126086.968 4748936.52 2816268.58 3557490.171 4957137.713 2027091.303 3933997.905 5092331.471 5488156.696 2958167.363 5946101.154 4239407.646 2805259.306 3420377.395 3312638.284 6086307.227 4564167.75 2058154.511 5944355.227 4456223.642 PE(P-16:0/20:4) 106832620.3 109574971.2 121129274.2 110306669.6 106440838.3 118637568.1 97423320.8 95752560.19 115746204.7 90396973.92 103490813.2 123473940.6 127422503.1 100484375.8 115838332.7 110541847.7 87944515.11 101009349.4 99307252.18 121873438.9 107650768.5 99682057.23 118640119.8 100800874.4 PE(P-16:0/20:5) 41823352.23 40892466.11 47613696.45 41729441.06 37977916.11 49517777.42 26425065.11 30684574.56 51525088.79 27962987.96 44362075.71 48368331.46 50504826.31 35348187.96 48030806.3 41531306.63 26046592.99 43167293.67 39445665.17 48384990.68 40510531.45 35825861.25 50952840.08 37857009.54 PE(P-16:0/22:4) 120909110.9 123410150.9 133787588.4 122675617.6 119212543.6 125439181.4 109425511.1 113501082.8 133024112.7 100029128.4 111939347.4 133316232.1 135867742.6 104202526 132743111.5 116697108.1 104363389.4 111764689.9 109685926.4 142863207.8 118151299.1 103923047.8 140189328 123615533.1 PE(P-16:0/22:6) 26735359.55 15246844.3 20219225.11 26409239.2 27889194.05 16836981.03 24467478.13 24280036.53 15476723.32 21721662.76 16107447.21 16721209.18 17341043.72 22766415.69 16444573.87 27044026.54 21056177.07 11965873.41 13301382.33 16108365.49 25178939.08 22885717.45 17999220.64 25865097.81 PE(P-18:0/18:1) 31321507.46 47538161.6 50967033.06 34503729.38 29088949.81 50106336.16 27331701.46 29304070.36 52668777.7 21762028.29 44939788.48 53169117.02 55682067.3 28125093.99 48959022.48 29302428.39 23106225.88 42755607.07 43084828.37 52986383.44 30725539.25 26444098.21 55139480.57 28851698.9 PE(P-18:0/18:2) 53212871.6 69025554.97 77140156.61 53836851.84 52274097.28 79011483.58 48990051.19 50356638.7 71933102.7 43256536.2 65400146.24 69715198.14 78881617.46 48420267.73 70021575.55 51093786.01 41982597.82 62219645.12 63463352.58 66074904.98 49969886.2 49622896.03 73475958.65 53566733.16 PE(P-18:0/20:4) 41541755.08 39303472.61 39617567.84 42096249.65 35707716.69 45794647.68 29776787.55 32621172.34 44882868.92 31649677.32 37919225.39 42007022.32 46492454.64 37241820.83 42834552.32 43981720.91 28651843.29 38482548.16 37165409.93 43954428.33 36922364.22 39355847.81 42039270.26 33181303.39 PE(P-18:0/20:5) 10708373.96 9913912.267 13123948.72 10806220.24 7559580.283 12020337.39 5459470.147 5741268.17 13436189.95 4940534.338 9420561.036 9894407.403 12453031.55 8601807.004 7201280.343 11491036.36 3425051.879 9601376.021 7801381.992 10136375.84 10733988.62 10645738.63 12070673.68 8586334.065 PE(P-18:0/22:6) 4618501.301 2590234.265 2508786.661 6140968.583 4522952.265 2320836.629 4270185.331 4341217.127 3742814.398 3714348.38 2183324.314 1994147.318 2846846.283 4344021.625 2309091.276 5309456.916 3190526.825 2296665.765 2154228.893 1970868.775 6285667.859 4289664.574 2766925.916 4264118.171 PE(P-18:1/20:4) 64532510.2 70972196.14 72066445.95 64133432.85 59805435.51 76796240.34 50451928.61 52684057.86 75090511.61 47721355.96 68225932.08 70974988.56 74761408.05 59558329.91 72904563.24 58721629.05 45630807.39 63973779.7 59346785.16 75413299.95 63963295.93 52670037.29 72787888.05 58507883.35 PE(P-18:1/22:6) 11735645.03 8001555.143 8683233.04 13674367.64 12876577.57 9283452.176 10660754.16 11223685.2 7823668.998 8848413.115 8771710.921 8567748.328 7378502.364 9922861.313 9165612.847 11340354.94 9202772.935 6602027.179 7633250.707 8460460.072 14779472.32 9493718.048 8464063.821 13139574.9 PEth(16:0_18:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PEth(18:1/18:1) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PG(15:0/18:1(d7)) 61546481.13 59314887.82 60283853.69 61999861.83 60127170.74 60966453.01 55200464.46 57926394.92 57993075.52 56851653.22 58923711.84 59794274.28 58401421.21 47424256.68 55060148.46 59526705.27 55096097.1 57589389.98 53790837.1 55779420.15 56781878.76 54512570.77 62365075.05 61127719.25 PG(16:0_18:1) 45557472.7 42162612.14 53566670.91 42666336.67 43150158.77 51773425.46 34762846.74 38820323.95 51978259.44 30428332.88 40882029.87 58153693.56 59996548.02 37381983.3 51073614.71 44928289.65 32407672.02 42161528.67 34432826.03 60288534.34 40784988.6 38940816.69 52796548.5 42371838.61 PG(16:0_20:4) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PG(16:0_22:6) 2315142.937 1739959.829 5841902.336 1474121.656 1192291.705 3463624.66 1114620.447 954173.3122 3876161.756 502239.5235 2763954.906 5653464.239 8873223.659 1146286.141 5560982.099 1413475.254 930518.3268 982331.8695 1926742.301 7786071.464 1703063.55 719576.1355 5506313.285 1022756.959 PG(18:0_18:1) 8829287.554 10768504.44 16189993.52 7595362.676 7770803.125 14205884.34 4816635.739 5685257.684 13408696.27 5165194.851 8574769.912 20864409.72 24443176.2 3430938.071 17291391.7 7646841.452 4584459.386 9078969.882 8655723.282 20654165.02 5552932.767 5108731.543 15158331.83 8677931.835 PG(18:0_20:4) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PG(18:0_22:6) 765621.0854 1121199.662 1689585.453 1135456.293 1159659.565 664618.8278 510864.0753 368650.2067 763820.4499 231780.5125 648625.1893 1474998.048 1763009.102 304977.5616 1253268.952 668091.3373 547218.6131 1025618.786 745229.4857 1286779.43 602308.1034 339793.8796 1188863.455 520978.5796 PG(18:1/18:1) 10151391.51 17484127.75 22112311.79 10882999.23 8749586.183 16543523.25 5999298.656 8444692.602 21156968.91 4589631.734 16852773.37 20742569.62 24267471.64 7142308.484 25036315.72 10206499.4 6555276.714 13606726.9 15036989.17 24296502.25 9737540.038 5779349.37 22830024.04 10093296.36 PI(15:0/18:1(d7)) 25242910.3 24917184.92 24740469.82 28195185.47 25581067.96 27565561.12 24088737.54 25349498.37 24891388.24 27138609.82 25130114.03 25093065.82 24860350.46 23197247.37 23640753.31 29782410.32 25388271.76 24068669.8 25534312.06 22531246.54 27844442.37 26846992.24 25582866.11 25409865.19 PI(16:0_18:1) 17527932.18 18846054.94 26100480.47 19945280.46 16416968.41 21563020.29 14235510.92 14123959.02 24827899.31 10481060.39 16540265.73 38246140.82 44754881.18 14581831.96 30925874.97 18694958.75 13797027.79 17064591.66 15893572.31 41700784.8 16615569.56 11842377.36 28382676.46 16811194.16 PI(16:0_20:4) 15439592.14 13796246.08 25299992.54 16191508.29 17227293.12 19369280.6 9006073.821 13863149.75 18903517.07 9536563.627 13368020.13 27416687.51 31838952.74 11157781.89 26014024.7 17598791.34 9746326.056 11872487.61 12698817.46 29694257.5 13932423.1 12081311.01 24559831.95 16232704.72 PI(16:0_22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PI(18:0_18:1) 3986842.642 8954023.52 11869534.86 4701525.344 5036689.274 10768282.4 3239453.955 4448842.912 12724779.77 2293392.216 8562726.495 21806412.68 21772177.26 3430463.137 12356347.24 3477086.966 3106415.703 6276113.397 5743126.457 21170124.16 5512512.242 3850307.814 13799072.21 5037518.866 PI(18:0_20:4) 49707982.12 57393655.31 67145123.34 53644873.38 51528363.84 59118188.33 46527060.48 51900275.67 59200527.93 42133311.02 54956758.11 57265871.23 60623892.5 47729858.09 63597459.64 52044054.05 44872071.67 51468470.52 51278088.93 58734416.89 51486495.86 44659380.7 62672022 53669582.81 PI(18:0_22:6) 1364250.341 1159893.84 1320168.945 1003639.45 1134618.619 1065004.095 928021.5188 1096191.739 1062500.229 865720.8038 1045856.276 1931612.315 2235962.054 459626.068 1361631.466 1020413.864 822858.6067 934182.1599 878000.9964 1846360.848 1378565.983 703159.9607 2162453.395 1356581.723 PI(18:1/18:1) 17164096.09 31083665.56 35909095.78 18199009.54 15449650.18 32357414.34 9861173.391 13481685.62 31288546.75 8167390.206 26899693.27 45026947.01 48845074.27 14435012.31 36492923.73 17088617.4 9171925.91 23541904.18 23855713.93 46141650.48 16036467.65 10757704.14 36741030.12 14344246.79 PI(20:4/20:4) 1517635.905 550271.1348 484222.7737 2127578.806 1366064.83 691877.0532 1238452.469 1232421.063 523690.3358 1223838.275 1159811.984 387239.9589 564248.0813 1973274.722 538861.7277 1632498.824 1414768.02 422144.889 400213.2817 454153.8152 2250258.241 1332027.782 544599.3418 2363908.234 PS(15:0/18:1(d7)) 30386995.56 30964224.19 31318616.74 31977934.33 34162247.41 30537873 28491922.65 33077533.74 33416408.19 34899626.1 30991238.51 33022971.05 33208848.51 26493457.23 28920196.34 35505276.88 32943948.82 31079541.62 31127870.92 27753350.22 30730657.79 25386030.08 36217537.11 33221949.73 PS(16:0_18:1) 99608454.84 103467922.1 142302150.5 107530100.8 99782784.17 121529637.5 77291884.5 89593050.01 118368284.2 74420157.56 104915072.7 144673476 162674932.7 90661025.68 143709462.1 100499226.9 80774027.82 101803606.8 104249628.9 162579322.4 101963832 85805147.75 153269196.3 102203856.9 PS(16:0_20:4) 7617051.473 5746159.765 11109400.35 5977996.841 5126791.341 7632431.664 2481328.705 4005226.102 7288154.764 2664526.704 5541812.37 15995383.55 20234241.98 4034809.654 9132804.702 4527626.932 1453603.172 5698087.373 3778137.446 13379074.96 5111942.304 4002449.474 12300733.7 4414883.853 PS(16:0_22:6) 7175512.319 2876910.24 6802791.582 4727585.865 6292043.32 5914403.874 2619974.904 4482423.745 4722529.331 2735450.789 4316112.046 9580607.951 9941760.2 3637860.487 7720776.282 6343683.733 3624207.078 3277818.196 1778574.403 10575065.59 4322162.854 4118498.199 5642475.791 6962066.991 PS(18:0_18:1) 98740036.89 120164215.6 116887072.3 83372675.68 91357789.06 121165492 92544284.03 107847176.4 116634433.7 74013633.19 114205365.4 165618005.5 153478879.6 76705202.17 111073528.4 101187344.2 78252376.46 87465564.02 96878948.93 132486586.7 85603728.55 74634478.94 117152846.4 104169565.9 PS(18:0_20:4) 22701279.94 29021105.05 35850897.55 26466930.04 21608010.7 33855981.22 14505326.48 16463692.25 35914916.98 12105211.37 27769776.9 45750563.76 52585800.88 20215429.59 37309783.09 26287683.68 14140243.86 23429982.48 24677088.05 48757800.3 28266783.2 20206534.62 44551095.55 21014483.05 PS(18:0_22:6) 27519747.54 21154948.53 30000964.87 27489268.2 27547785.41 29332531.6 19107670.52 23849763.3 29444649.27 20627386.68 20602030.66 38372223.53 42824422.56 22231203.56 34726372.49 26031230.33 20896000.52 19807935.02 20981397 41546693.13 26378032.69 23393185.94 37211319.67 27280832.76 PS(18:1/18:1) 50034972.62 72132330.98 87432440.38 53889756.46 43775810 82911583.68 35850964.74 39187793.67 79950755.9 28611734.57 65958381.55 91099661.92 100794416.2 39761441.23 94423615.04 41513204.41 32811260.67 62564108.08 64876610.22 92825037.01 49338449.28 39686432.43 96332919.95 39306907.91 PS(22:6/22:6) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Stearic acid NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA PE(P-18:0/18:1(d9)) NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name Expected RT MS1_mz MS2_mz (3-O-sulfo)GalCer(d18:1/16:0) 4.45 778.5 97 (3-O-sulfo)GalCer(d18:1/18:0(2OH) 4.87 822.6 97 (3-O-sulfo)GalCer(d18:1/18:0(d3)) 5.06 809.6 97 (3-O-sulfo)GalCer(d18:1/18:0) 4.96 806.6 97 (3-O-sulfo)GalCer(d18:1/24:0(2OH)) 6.14 906.7 97 (3-O-sulfo)GalCer(d18:1/24:0) 6.19 890.7 97 (3-O-sulfo)GalCer(d18:1/24:1(2OH)) 5.73 904.7 97 (3-O-sulfo)GalCer(d18:1/24:1) 5.79 888.7 97 Arachidonic acid 2.33 303.2 303.2 Arachidonic acid_MRM 2.33 303.2 259.1 Arachidonic acid-d8 2.3 311.3 311.3 Arachidonic acid-d8_MRM 2.3 311.3 267.1 BMP(14:0/14:0) 3.69 665.3 227.2 BMP(16:0_18:1) 4.8 747.5 255.4 BMP(16:0_20:4) 4.13 769.5 255.4 BMP(16:0_22:6) 4.2 795.5 255.4 BMP(16:1/16:1) 3.79 717.5 253.1 BMP(18:0_18:1) 5.25 775.5 281.4 BMP(18:0_20:4) 4.69 797.5 283.4 BMP(18:0_22:6) 4.7 823.5 283.4 BMP(18:1/18:1) 4.84 773.5 281.3 BMP(20:4/20:4) 3.69 817.5 303.3 BMP(22:6/22:6) 3.45 865.5 327.3 Cholesterol sulfate 3.08 465.3 96.7 CL(14:0/14:0/14:0/14:0) 7.17 619.5 227.2 CL(72:6/18:2) 7.48 725.7 279.2 CL(72:7/18:2) 7.24 724.7 279.2 CL(72:8-2(OOH)/18:2) 6.49 755.7 279.2 CL(72:8/18:2) 7.26 723.7 279.3 CL(74:9/18:2) 7.29 736.7 279.2 DHA 2.14 327.2 229.1 EPA 1.97 301.3 257.1 FAHFA(18:1/9-O-18:0) 6.2 563.6 281 GD1a/b(d36:1) 4.54 917.5 290.1 GD3(d34:1) 4.1 720.9 290.1 GD3(d36:1) 4.65 734.9 290.1 GM3(d18:1/18:0(d5)) 5.03 1184.8 290.1 GM3(d34:1) 4.54 1151.7 290.1 GM3(d36:1) 5.05 1179.8 290.1 GQ1b(d36:1) 4.13 1208.6 290.1 Hemi-BMP(14:0/14:0)_14:0 6.2 875.5 227.3 Hemi-BMP(18:1/18:1)_16:0 7.1 1011.7 281.3 Hemi-BMP(18:1/18:1)_18:0 7.2 1039.7 281.3 Hemi-BMP(18:1/18:1)_18:1 7.1 1037.7 281.3 Hemi-BMP(20:4/20:4)_16:0 6.4 1056.8 303.3 Hemi-BMP(20:4/20:4)_18:0 6.6 1185.8 303.3 Hemi-BMP(20:4/20:4)_18:1 6.4 1183.8 303.3 Hemi-BMP(20:4/20:4)_20:4 6.2 1104.8 303.3 Hemi-BMP(22:6/22:6)_16:0 6.2 1103.7 327.3 Hemi-BMP(22:6/22:6)_18:0 6.5 1131.7 327.3 Hemi-BMP(22:6/22:6)_18:1 6.2 1129.7 327.3 Hemi-BMP(22:6/22:6)_22:6 5.6 1175.7 327.3 Linoleic acid 2.46 279.2 279.2 Linolenic acid 2.08 277.2 277.2 LPE(16:0) 1.97 452.2 255.3 LPE(18:0) 2.61 480.3 283.3 LPE(18:1(d7)) 2 485.3 288.3 LPG(16:0) 1.65 483.3 255.3 LPG(18:0) 2.11 511.3 283.3 LPG(18:1) 1.64 509.3 281.3 LPG(20:4) 1.29 531.3 303.3 LPG(22:6) 1.28 555.3 327.3 LPI(16:0) 1.6 571.3 241.1 LPI(18:0) 2.12 599.3 241.1 Oleic acid 2.96 281.2 281.2 PA(15:0/18:1(d7)) 5.09 666.5 241.3 PA(16:0_18:1) 5.45 673.5 255.3 PA(18:0_18:1) 5.84 701.5 283.3 PA(18:0_20:4) 5.42 723.5 283.3 PA(18:0_22:6) 5.35 747.5 283.3 PA(18:1/18:1) 5.36 699.5 281.3 Palmitic acid 2.98 255.1 255.1 Palmitoleic acid 2.35 253.1 253.1 PE(15:0/18:1(d7)) 5.49 709.6 241.3 PE(O-16:0/20:4) 5.48 724.5 303.2 PE(O-16:0/22:6) 5.38 748.5 327.2 PE(O-18:0/20:4) 5.97 752.6 303.2 PE(O-18:0/22:6) 5.87 776.6 327.2 PE(P-16:0/20:4) 5.48 722.6 303.3 PE(P-16:0/20:5) 5.11 720.6 301.3 PE(P-16:0/22:4) 5.83 750.6 331.3 PE(P-16:0/22:6) 5.29 746.6 327.3 PE(P-18:0/18:1) 6.4 728.6 281.3 PE(P-18:0/18:2) 6.08 726.6 279.2 PE(P-18:0/20:4) 5.96 750.6 303.3 PE(P-18:0/20:5) 5.61 748.6 301.3 PE(P-18:0/22:6) 5.79 774.6 327.3 PE(P-18:1/20:4) 5.48 748.5 303.3 PE(P-18:1/22:6) 5.29 772.5 327.3 PEth(16:0_18:1) 5.4 772.5 255.1 PEth(18:1/18:1) 5.4 773.6 281.2 PG(15:0/18:1(d7)) 4.87 740.6 241.3 PG(16:0_18:1) 5.13 747.5 255.3 PG(16:0_20:4) 4.61 769.5 255.3 PG(16:0_22:6) 4.28 795.5 255.3 PG(18:0_18:1) 5.6 775.5 281.3 PG(18:0_20:4) 5.14 797.5 283.3 PG(18:0_22:6) 4.79 823.5 283.3 PG(18:1/18:1) 5.14 773.4 281.4 PI(15:0/18:1(d7)) 4.87 828.6 241.3 PI(16:0_18:1) 5.04 835.6 255.3 PI(16:0_20:4) 4.61 857.6 255.3 PI(16:0_22:6) 4.43 881.6 255.3 PI(18:0_18:1) 5.51 863.6 283.3 PI(18:0_20:4) 5.12 885.6 283.3 PI(18:0_22:6) 4.97 909.6 283.3 PI(18:1/18:1) 5.04 861.6 281.3 PI(20:4/20:4) 3.96 905.6 303.3 PS(15:0/18:1(d7)) 4.94 753.6 241.3 PS(16:0_18:1) 5.19 760.6 255.3 PS(16:0_20:4) 4.68 782.6 255.3 PS(16:0_22:6) 4.5 806.6 255.3 PS(18:0_18:1) 5.66 788.6 283.3 PS(18:0_20:4) 5.25 810.6 283.3 PS(18:0_22:6) 5.02 834.6 283.3 PS(18:1/18:1) 5.19 786.6 281.3 PS(22:6/22:6) 3.66 878.5 327.3 Stearic acid 3.6 283.2 283.2 PE(P-18:0/18:1(d9)) 6.4 737.6 290.2 METABOLITES_END #END