#METABOLOMICS WORKBENCH jdcolter_20250531_102131 DATATRACK_ID:5978 STUDY_ID:ST004070 ANALYSIS_ID:AN006736 PROJECT_ID:PR002556 VERSION 1 CREATED_ON July 18, 2025, 10:58 am #PROJECT PR:PROJECT_TITLE Metabolic perturbation in ependymal cells leads to local and distant PR:PROJECT_TITLE neurodegeneration and cognitive decline PR:PROJECT_TYPE MS Imaging Analysis PR:PROJECT_SUMMARY Ependymal cells (ECs) are specialized multi-ciliated glial cells that line the PR:PROJECT_SUMMARY ventricular system of the brain, regulating cerebrospinal fluid flow (CSF) and PR:PROJECT_SUMMARY the neighbouring neural stem cell (NSC) niche. However, their role in PR:PROJECT_SUMMARY maintaining brain homeostasis or in disease pathogenesis remains unclear. To PR:PROJECT_SUMMARY elucidate their function, we disrupted ependymal glucose metabolism by PR:PROJECT_SUMMARY genetically deleting glucose-transporter-1 (GLUT1/Slc2a1) in postnatal ECs. PR:PROJECT_SUMMARY Analyses were carried out across three separate studies (batches), with one PR:PROJECT_SUMMARY study at 1 month (6 mice) and two studies at 12 months (3 mice, 5 mice). Results PR:PROJECT_SUMMARY from this project confirm CSF flow changes and disrupted NSC differentiation and PR:PROJECT_SUMMARY neuroblast migration. These mice also exhibited periventricular lipid droplet PR:PROJECT_SUMMARY accumulation similar to Alzheimer’s disease brains. Aged cKO mice exhibited PR:PROJECT_SUMMARY progressive cognitive and motor dysfunction, and onset of seizure activity. PR:PROJECT_SUMMARY These behavioral deficits were coincident with various neurodegenerative PR:PROJECT_SUMMARY pathologies, including dysmyelination, microglia-associated inflammation, and PR:PROJECT_SUMMARY lipid imbalance. When combined with metabolic perturbation in ECs, 5xFAD mice PR:PROJECT_SUMMARY exhibited accelerated disease onset. These findings suggest that ECs are PR:PROJECT_SUMMARY important regulators of brain homeostasis, and their dysfunction may contribute PR:PROJECT_SUMMARY to the pathogenesis of neurodegenerative diseases. PR:INSTITUTE University of Calgary PR:DEPARTMENT Veterinary Medicine PR:LABORATORY Biernaskie Lab PR:LAST_NAME Colter PR:FIRST_NAME James PR:ADDRESS 2500 University Drive NW PR:EMAIL jdcolter@ucalgary.ca PR:PHONE +1 (403) 210-7306 PR:FUNDING_SOURCE CIHR PR:PROJECT_COMMENTS Part 1 of 3 PR:PUBLICATIONS (Under Review) PR:CONTRIBUTORS Nilesh Sharma, Alexander Pun, James Colter, Leslie Cao, Nicole Rosin, Dominic PR:CONTRIBUTORS Gerding, Isabel Rea, Apolline Pistek, Qandeel Shafqat, Sarthak Sinha, Elodie PR:CONTRIBUTORS Labit, Eren Kutluberk, Caleb Small, Reese Landes, Tak Ho Chu, Kartikeya Murari, PR:CONTRIBUTORS E. Dale Abel, Jeffrey T. Joseph, Rehana Leak, Jeff Dunn, and Jeff Biernaskie #STUDY ST:STUDY_TITLE Metabolic perturbation in ependymal cells leads to local and distant ST:STUDY_TITLE neurodegeneration and cognitive decline - Study 1 of 3 (12-month Glut1KO against ST:STUDY_TITLE Ctrl) ST:STUDY_SUMMARY Ependymal cells (ECs) are specialized multi-ciliated glial cells that line the ST:STUDY_SUMMARY ventricular system of the brain, regulating cerebrospinal fluid flow (CSF) and ST:STUDY_SUMMARY the neighbouring neural stem cell (NSC) niche. However, their role in ST:STUDY_SUMMARY maintaining brain homeostasis or in disease pathogenesis remains unclear. To ST:STUDY_SUMMARY elucidate their function, we disrupted ependymal glucose metabolism by ST:STUDY_SUMMARY genetically deleting glucose-transporter-1 (GLUT1/Slc2a1) in postnatal ECs. ST:STUDY_SUMMARY Analyses were carried out across three separate studies (batches), with one ST:STUDY_SUMMARY study at 1 month (6 mice) and two studies at 12 months (3 mice, 5 mice). Results ST:STUDY_SUMMARY from this project confirm CSF flow changes and disrupted NSC differentiation and ST:STUDY_SUMMARY neuroblast migration. These mice also exhibited periventricular lipid droplet ST:STUDY_SUMMARY accumulation similar to Alzheimer’s disease brains. Aged cKO mice exhibited ST:STUDY_SUMMARY progressive cognitive and motor dysfunction, and onset of seizure activity. ST:STUDY_SUMMARY These behavioral deficits were coincident with various neurodegenerative ST:STUDY_SUMMARY pathologies, including dysmyelination, microglia-associated inflammation, and ST:STUDY_SUMMARY lipid imbalance. When combined with metabolic perturbation in ECs, 5xFAD mice ST:STUDY_SUMMARY exhibited accelerated disease onset. These findings suggest that ECs are ST:STUDY_SUMMARY important regulators of brain homeostasis, and their dysfunction may contribute ST:STUDY_SUMMARY to the pathogenesis of neurodegenerative diseases. In this part, 3x mice were ST:STUDY_SUMMARY analyzed at 12-months under Glut1ko (2 mice) or Control (1 mouse) conditions. ST:STUDY_SUMMARY See study 2 for additional replicates. ST:INSTITUTE University of Calgary ST:DEPARTMENT Veterinary Medicine ST:LABORATORY Biernaskie Lab ST:LAST_NAME Colter ST:FIRST_NAME James ST:ADDRESS 2500 University Drive NW, Calgary AB Canada, T2N1N4 ST:EMAIL jdcolter@ucalgary.ca ST:PHONE +1 (403) 210-7306 ST:NUM_GROUPS 4 ST:TOTAL_SUBJECTS 13 ST:STUDY_COMMENTS Part 1 of 3 ST:PUBLICATIONS (under review) #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:AGE_OR_AGE_RANGE 12 months #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 220809_fourbrains_mets 565 Condition:Control | Sample source:Brain Age (Months)=12; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_1.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_2.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_3.mcf SUBJECT_SAMPLE_FACTORS 220809_fourbrains_mets 271 Condition:Knockout | Sample source:Brain Age (Months)=12; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_1.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_2.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_3.mcf SUBJECT_SAMPLE_FACTORS 220809_fourbrains_mets 373 Condition:Knockout | Sample source:Brain Age (Months)=12; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_1.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_2.mcf; RAW_FILE_NAME(Raw Data Files)=a2a57bd9-28ce-4016-936b-be1c11e9ad24_3.mcf #COLLECTION CO:COLLECTION_SUMMARY The brain was collected as mentioned below. After the mice were euthanized, it CO:COLLECTION_SUMMARY was transcardially perfused with PBS. The brain is dissected out and then flash CO:COLLECTION_SUMMARY frozen in liquid nitrogen. The flash frozen brain tissue was then sectioned at CO:COLLECTION_SUMMARY 12 μm thickness on a cryostat (Leica Biosystems). CO:SAMPLE_TYPE Brain #TREATMENT TR:TREATMENT_SUMMARY The brain was collected as mentioned above with minor modifications. After the TR:TREATMENT_SUMMARY mice were euthanized, it was transcardially perfused with PBS. The brain is TR:TREATMENT_SUMMARY dissected out and then flash frozen in liquid nitrogen. The flash frozen brain TR:TREATMENT_SUMMARY tissue was then sectioned at 12 μm thickness on a cryostat (Leica Biosystems). TR:TREATMENT_SUMMARY MALDI matrix (9-aminoacridine, 9AA) (Sigma-Aldrich) was spray-coated onto the TR:TREATMENT_SUMMARY target slides in an automated fashion using a TM Sprayer (HTX Imaging). 9-AA was TR:TREATMENT_SUMMARY made up as a 5 mg/ml solution in 90% methanol. Four passes were used with a TR:TREATMENT_SUMMARY nozzle temperature of 85°, a flowrate of 0.15 ml/min, 2-mm track spacing, and a TR:TREATMENT_SUMMARY stage velocity of 700 mm/min. Nitrogen was used as the nebulization gas and was TR:TREATMENT_SUMMARY set to 10 psi. Images were acquired on a 15T Fourier transform ion cyclotron TR:TREATMENT_SUMMARY resonance mass spectrometer (FT-ICR MS, Solarix, Bruker Daltonics) equipped with TR:TREATMENT_SUMMARY an Apollo II dual ion source and Smartbeam II 2kHz Nd:YAG laser that was TR:TREATMENT_SUMMARY frequency tripled to 355 nm. Data were collected in the negative ion mode with TR:TREATMENT_SUMMARY the laser operating at 2 kHz at 50 μm resolution. Tentative metabolite TR:TREATMENT_SUMMARY identifications were made by accurate mass, typically better than 1 ppm. Images TR:TREATMENT_SUMMARY were analyzed with flexImaging software (Bruker), while average spectra were TR:TREATMENT_SUMMARY exported to mMass for visualization of differences. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY MALDI matrix (9-aminoacridine, 9AA) (Sigma-Aldrich) was spray-coated onto the SP:SAMPLEPREP_SUMMARY target slides in an automated fashion using a TM Sprayer (HTX Imaging). 9-AA was SP:SAMPLEPREP_SUMMARY made up as a 5 mg/ml solution in 90% methanol. Four passes were used with a SP:SAMPLEPREP_SUMMARY nozzle temperature of 85°, a flowrate of 0.15 ml/min, 2-mm track spacing, and a SP:SAMPLEPREP_SUMMARY stage velocity of 700 mm/min. Nitrogen was used as the nebulization gas and was SP:SAMPLEPREP_SUMMARY set to 10 psi. Images were acquired on a 15T Fourier transform ion cyclotron SP:SAMPLEPREP_SUMMARY resonance mass spectrometer (FT-ICR MS, Solarix, Bruker Daltonics) equipped with SP:SAMPLEPREP_SUMMARY an Apollo II dual ion source and Smartbeam II 2kHz Nd:YAG laser that was SP:SAMPLEPREP_SUMMARY frequency tripled to 355 nm. Data were collected in the negative ion mode with SP:SAMPLEPREP_SUMMARY the laser operating at 2 kHz at 50 μm resolution. Tentative metabolite SP:SAMPLEPREP_SUMMARY identifications were made by accurate mass, typically better than 1 ppm. Images SP:SAMPLEPREP_SUMMARY were analyzed with flexImaging software (Bruker), while average spectra were SP:SAMPLEPREP_SUMMARY exported to mMass for visualization of differences. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE None (Direct infusion) CH:INSTRUMENT_NAME none CH:COLUMN_NAME none CH:SOLVENT_A none CH:SOLVENT_B none CH:FLOW_GRADIENT none CH:FLOW_RATE none CH:COLUMN_TEMPERATURE none #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Bruker Solarix FT-ICR-MS MS:INSTRUMENT_TYPE MALDI-TOF MS:MS_TYPE MALDI MS:ION_MODE NEGATIVE MS:MS_COMMENTS Methods below. Please note that the processed data files included in this MS:MS_COMMENTS submission only provide the identified metabolites in the set, since the data MS:MS_COMMENTS format for workbench does not allow a 3-dimensional approach (each pixel MS:MS_COMMENTS contains a spectrum, and sample brains within an experiment are sectioned by MS:MS_COMMENTS brain region and analyzed by metabolite across spatial coordinates). Please MS:MS_COMMENTS refer to github.com/BiernaskieLab for .csv files containing spectral intensities MS:MS_COMMENTS by metabolite across spatial coordinates for specific brain regions by sample. MS:MS_COMMENTS MALDI-TOF spectral data was acquired using Bruker Compass FlexImaging software. MS:MS_COMMENTS The raw dataset was exported to Bruker SciLS Lab for peak-finding, isolation of MS:MS_COMMENTS spectral data by region. Reorganized data was exported as .csv files for MS:MS_COMMENTS cross-dataset comparisons utilizing Python. Total ion count (TIC) normalization MS:MS_COMMENTS was applied prior to exporting numerical intensity data from SciLS Lab or MS:MS_COMMENTS plotting and exporting images from FlexImaging. Automated peak-finding was MS:MS_COMMENTS applied within Bruker SciLS Lab to TIC-normalized datasets to generate an ion MS:MS_COMMENTS list for further processing. This ion list was cross-referenced between datasets MS:MS_COMMENTS to harmonize the ion list across brain sections imaged in all experiments. The MS:MS_COMMENTS COMP_DB database at https://www.lipidmaps.org was applied to annotate ions with MS:MS_COMMENTS potential lipid hits. A delta m/z of ±0.01 was used. If no matches were MS:MS_COMMENTS returned, this delta was increased to ±0.02, and repeated at ±0.05 if nothing MS:MS_COMMENTS was returned. Ions that had no matches in the database beyond ±0.05 were MS:MS_COMMENTS excluded from the set. Given the limitations of assessing fragmentation profiles MS:MS_COMMENTS of the mass spectral data, ions with potential matches from multiple lipid MS:MS_COMMENTS groups were classified as ‘unknown’. Those ions with multiple possible MS:MS_COMMENTS matches from the same group were included in their respective lipid group and MS:MS_COMMENTS annotated with all possible matches. This resulted in a list of 48 ions with MS:MS_COMMENTS known lipid groups. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS n/a MS_METABOLITE_DATA_START Samples 565 271 373 Factors Condition:Control | Sample source:Brain Condition:Knockout | Sample source:Brain Condition:Knockout | Sample source:Brain CAR 13:0;O4 1 1 1 CerP 34:0;O6 1 1 1 SHexCer 28:2;O4 1 1 1 SHexCer 30:5;O3 1 1 1 FA 16:0 1 1 1 Cer 34:0;O, NAE 32:0 1 1 1 FA 15:4;O2 1 1 1 FA 18:2 1 1 1 Cer 37:1;O 1 1 1 FA 18:1 1 1 1 Cer 37:0;O 1 1 1 NAE 15:1, SPB 17:2;O2 1 1 1 Cer 36:0;O2, NAE 34:0;O 1 1 1 FA 17:1;O, MG O-14:2 1 1 1 FA 18:0 1 1 1 PIP 30:4 1 1 1 NAE 15:0, SPB 17:1;O2 1 1 1 Cer 38:4;O 1 1 1 Cer 38:2;O 1 1 1 ST 21:5;O8;GlcA 1 1 1 CAR 27:1;O3, Cer 34:2;O6, DGTA 24:0, DGTS 24:0 1 1 1 CAR 28:0;O2, Cer 35:1;O5, NAE 33:1;O4 1 1 1 CerP 33:1;O2, LPC O-25:2, LPE O-28:2, CAR 26:0;O4 1 1 1 CAR 26:0;O4 1 1 1 Cer 34:0;O6 1 1 1 NAT 34:4 1 1 1 CAR 30:2;O, Cer 37:3;O4, LPC O-25:0, LPE O-28:0 1 1 1 ST 18:3;O4 1 1 1 FA 20:4, ST 20:1;O2 1 1 1 NAE 17:4 1 1 1 CoA 10:0 1 1 1 FA 13:3;O4 1 1 1 ST 25:7;O7;GlcA 1 1 1 FA 20:1 1 1 1 HexCer 28:1;O3 1 1 1 CerP 33:1;O4 1 1 1 FA 22:6, ST 22:3;O2 1 1 1 CAR 11:0, NAE 16:1;O2, SPB 18:2;O4, NAE 15:1, SPB 17:2;O2 1 1 1 FA 22:4, ST 22:1;O2 1 1 1 NAE 19:4 1 1 1 ACer 65:0;O4 1 1 1 ACer 64:3;O6, DGCC 54:2 1 1 1 PIP3 31:7;O 1 1 1 ST 18:5;O2;S 1 1 1 ST 18:5;O5 1 1 1 SHexCer 53:0;O4 1 1 1 PC 56:0;O, PE 59:0;O, PS O-58:0, PT O-57:0 1 1 1 SHexCer 29:6;O4 1 1 1 SHexCer 28:5;O5 1 1 1 PA O-66:1 1 1 1 ACer 74:2;O2 1 1 1 PIP 22:4 1 1 1 CoA 24:2 1 1 1 TG 68:2;O2, TG O-68:3;O3 1 1 1 ST 18:3;O6 1 1 1 PIP2 43:12;O, PIP3 36:3;O 1 1 1 NAT 11:2;O3 1 1 1 CoA 23:0;O 1 1 1 TG 70:5;O, TG O-70:6;O2 1 1 1 HexCer 36:6;O4 1 1 1 FA 20:2;O2, MG 17:2, MG O-17:3;O 1 1 1 Hex2Cer 50:1;O5 1 1 1 CoA 22:5;O4 1 1 1 PIP 24:5 1 1 1 LPA O-13:3 1 1 1 TG 73:1, TG O-73:2;O 1 1 1 CoA 24:5;O3 1 1 1 SPBP 15:3;O3, M(IP)2C 28:5;O6 1 1 1 ST 18:4;O7 1 1 1 ST 19:3;O6 1 1 1 ST 22:6;O;S 1 1 1 CoA 27:7;O 1 1 1 NAT 11:0;O3 1 1 1 PIP 23:2;O, PIP3 41:6 1 1 1 LPA O-16:2 1 1 1 TG O-46:10;O3 1 1 1 SQDG 33:2, FA 22:3;O2, MG 19:3, MG O-19:4;O, ST 22:0;O4 1 1 1 TG 75:7;O2, TG O-75:8;O3 1 1 1 LPE 10:2, SPBP 15:3;O4 1 1 1 ST 22:6;O2;S 1 1 1 NAT 10:0;O4, NAT 11:0;O4 1 1 1 DGCC 68:10 1 1 1 FA 22:2;O2, MG 19:2, MG O-19:3;O, SQDG 33:0 1 1 1 MGDG 67:3 1 1 1 TG 75:6;O3 1 1 1 TG O-81:14, MGDG 68:4 1 1 1 LPA 16:0, LPA O-16:1;O 1 1 1 LPA O-18:3, FA 22:3;O3, MG 19:3;O, ST 22:0;O5 1 1 1 CAR 13:1;O4 1 1 1 CAR 15:1, NAE 20:2;O2, MGDG 41:9 1 1 1 LPA O-18:2, FA 22:2;O3, MG 19:2;O 1 1 1 SQDG 66:4, TG 82:18;O, TG O-82:19;O2 1 1 1 CAR 13:0;O4, CAR 12:0;O2, NAE 17:1;O4, CAR 11:0;O2, NAE 16:1;O4 1 1 1 TG 81:10;O2, TG O-81:11;O3 1 1 1 LPA 18:1, LPA O-18:2;O 1 1 1 LPC O-13:1, LPE O-16:1 1 1 1 FA 22:1;O4, LPA 18:0, LPA O-18:1;O 1 1 1 NAT 22:4 1 1 1 DG 21:5, DG O-21:6;O, FA 24:6;O3, MG 21:6;O, ST 24:3;O5 1 1 1 PI 68:1;O 1 1 1 SHexCer 37:0;O6 1 1 1 CAR 17:2, NAE 22:3;O2 1 1 1 PA 49:6;O, PEth 47:6;O, PG O-46:7 1 1 1 HexCer 45:6;O5 1 1 1 CAR 17:1, NAE 22:2;O2, MGDG 45:9 1 1 1 TG 87:19;O2, TG O-87:20;O3 1 1 1 ST 19:3;O5;S 1 1 1 LPC 13:0, LPC O-13:1;O, LPE 16:0, LPE O-16:1;O 1 1 1 LPC O-15:2, LPE O-18:2 1 1 1 ST 26:5;O5 1 1 1 LPC O-15:1, LPE O-18:1, CAR 17:0;O, NAE 22:1;O3 1 1 1 ST 20:3;O4;T 1 1 1 LPC 15:1, LPC O-15:2;O, LPE 18:1, LPE O-18:2;O 1 1 1 LPC 15:0, LPC O-15:1;O, LPE 18:0, LPE O-18:1;O, CAR 17:0;O2, NAE 22:1;O4 1 1 1 PI O-45:7 1 1 1 LPC 16:2;O, LPE 19:2;O, LPS O-18:2, LPT O-17:2, LPC O-15:3, LPE O-18:3 1 1 1 ST 28:7;O6;S, ST 19:2;O5;GlcA, ST 19:3;O6;Hex 1 1 1 PE 23:6, ST 27:4;O6;G, PC 21:6 1 1 1 ACer 73:0;O4 1 1 1 ST 18:1;O7;GlcA, ST 18:2;O8;Hex 1 1 1 BMP 21:1;O, LPI O-18:2, PG 21:1;O 1 1 1 LPI 18:0, LPI O-18:1;O, BMP 20:0, LPG 20:1;O, PG 20:0, PG O-20:1;O 1 1 1 ST 30:0;O8;T, NAT 29:3;O4, ST 29:0;O6;T 1 1 1 PIP2 56:2 1 1 1 LPA 34:2;O, LPG O-31:3, PA 34:1, PA O-34:2;O 1 1 1 HexCer 30:1;O4, CAR 28:1;O4, DGCC 25:0, HexCer 29:1;O2, CAR 27:1;O4 1 1 1 LPG O-33:4, PA 36:2, PA O-36:3;O, LPA O-34:4 1 1 1 HexCer 32:2;O4, CAR 29:2;O4, DGCC 26:1, HexCer 30:2;O2 1 1 1 CerP 39:2;O3, LPC 31:2, LPC O-31:3;O 1 1 1 LPG O-33:3, PA 36:1, PA O-36:2;O, PEth 34:1, LPA O-34:3 1 1 1 HexCer 32:1;O4, CAR 29:1;O4, DGCC 26:0, HexCer 30:1;O2 1 1 1 CerPE 38:1;O2 1 1 1 CerP 39:2;O4, LPC 31:2;O, DGTA 30:1 1 1 1 Cer 45:6;O2 1 1 1 CerP 39:1;O4, LPC 31:1;O, PC 32:0 1 1 1 CE 17:1;O3, DG 41:5 1 1 1 CerP 41:5;O3, LPC 33:5 1 1 1 PA 38:4, DG 39:5;O2, TG 39:4;O 1 1 1 CerP 41:3;O3, LPC 33:3, LPC O-33:4;O, PE O-36:3 1 1 1 TG 94:21;O3, CE 19:3;O2, DG O-43:8 1 1 1 TG 94:20;O3, PC O-34:2, CerP 41:2;O3, LPC 33:2 1 1 1 CerP 41:3;O4, LPC 33:3;O, PE 36:2 1 1 1 CerP 41:2;O4, LPC 33:2;O, PE 36:1 1 1 1 PA 40:7, PA O-40:8;O, PEth 38:7, DG 41:8;O2, TG 41:7;O, TG O-41:8;O2 1 1 1 CL 74:0 1 1 1 PE O-38:7 1 1 1 PA 40:6, PA O-40:7;O, PEth 38:6, DG 41:7;O2, TG 41:6;O 1 1 1 DG O-45:12 1 1 1 HexCer 36:6;O4, CAR 34:6;O4, DGCC 31:5 1 1 1 CerP 43:6;O3, PE O-38:6, PC O-36:6 1 1 1 CerPE 37:3;O3, SM 34:3;O3 1 1 1 CerP 43:5;O3, PE O-38:5 1 1 1 PA 40:4, TG 41:4;O, DG 41:5;O2 1 1 1 CE 21:5;O2, DG O-45:10 1 1 1 CerP 43:3;O3, PE O-38:3, PC O-36:3 1 1 1 PE 38:6, PE O-38:7;O, DGTA 34:6, PC 36:6 1 1 1 CL 78:10 1 1 1 CerP 43:6;O4, PE 38:5, LPC O-34:7 1 1 1 CerP 43:5;O4, PE 38:4, DGTA 34:4, LPC O-34:6 1 1 1 CE 21:5;O3, DG 45:9, CL 78:6 1 1 1 PE O-40:8, CerP 39:0;O5, LPS O-33:0;O, PC O-38:8 1 1 1 CerP 43:2;O4, PE 38:1, LPC O-34:3, PC 36:1 1 1 1 PE O-40:7, PC O-38:7 1 1 1 PG 36:1, PG O-36:2;O 1 1 1 CerP 45:6;O3, PE O-40:6, PC O-38:6 1 1 1 CerP 45:5;O3, PE O-40:5, PC O-38:5 1 1 1 PE 40:7, PE O-40:8;O 1 1 1 CerP 42:3;O6, PS 36:1, PT 35:1, LPC 33:3;O 1 1 1 PA 43:6, MGDG 37:5, TG 41:5;O2 1 1 1 PE 40:6, PC 38:6 1 1 1 CL 82:10 1 1 1 Hex2Cer 29:0;O2 1 1 1 CerP 45:5;O4, PE 40:4, PC 38:4 1 1 1 SHexCer 36:1;O2 1 1 1 PS 40:6, PT 39:6, PC O-37:8;O, PE 40:7 1 1 1 LPI 34:2;O, PG 35:2;O 1 1 1 PC O-39:8, PE O-42:8 1 1 1 SHexCer 38:1;O3 1 1 1 IPC 36:2;O3 1 1 1 PI 36:4, PG 38:5;O 1 1 1 PC O-41:11, PE O-44:11 1 1 1 SHexCer 40:2;O2 1 1 1 CerP 47:3;O4, PC 39:2, PE 42:2 1 1 1 CerPE 42:2;O5, SM 39:2;O5 1 1 1 CerP 47:4;O5, PC 39:3;O, PE 42:3;O 1 1 1 SHexCer 41:1;O2 1 1 1 HexCer 43:6;O6 1 1 1 CerPE 42:2;O6, SM 39:2;O6 1 1 1 PI 38:5, PI O-38:6;O 1 1 1 PC O-43:12, PE O-46:12, Hex2Cer 34:5;O4 1 1 1 PI 38:4, PG 40:5;O 1 1 1 PC O-43:11, PE O-46:11, Hex2Cer 34:4;O4 1 1 1 SHexCer 42:3;O2 1 1 1 PG 42:5, PG O-42:6;O 1 1 1 SM 41:4;O5 1 1 1 SHexCer 42:2;O2 1 1 1 TG 52:10;O3, CerPE 44:3;O5, SM 41:3;O5 1 1 1 SHexCer 41:2;O3 1 1 1 SHexCer 42:1;O2 1 1 1 CerPE 43:2;O5, SM 40:2;O5 1 1 1 CerP 48:3;O5, PC 40:2;O, PE 43:2;O, PS O-42:2, PT O-41:2 1 1 1 CerPE 43:2;O6, SM 40:2;O6 1 1 1 SHexCer 42:3;O3 1 1 1 PC 41:3;O, PE 44:3;O, PS O-43:3, PT O-42:3 1 1 1 CerPE 44:3;O6, SM 41:3;O6 1 1 1 CerP 49:3;O5, PC 41:2;O, PE 44:2;O, PS O-43:2, PT O-42:2 1 1 1 CerPE 44:2;O6, SM 41:2;O6 1 1 1 TG 86:7 1 1 1 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name mz CAR 13:0;O4 209.62863 CerP 34:0;O6 226.8164 SHexCer 28:2;O4 240.79327 SHexCer 30:5;O3 242.79022 FA 16:0 255.23307 Cer 34:0;O, NAE 32:0 260.76124 FA 15:4;O2 265.14801 FA 18:2 279.23291 Cer 37:1;O 280.78411 FA 18:1 281.24867 Cer 37:0;O 281.78907 NAE 15:1, SPB 17:2;O2 282.25202 Cer 36:0;O2, NAE 34:0;O 282.7812 FA 17:1;O, MG O-14:2 283.24318 FA 18:0 283.26421 PIP 30:4 283.78609 NAE 15:0, SPB 17:1;O2 284.24652 Cer 38:4;O 284.77483 Cer 38:2;O 286.77199 ST 21:5;O8;GlcA 290.08821 CAR 27:1;O3, Cer 34:2;O6, DGTA 24:0, DGTS 24:0 298.72943 CAR 28:0;O2, Cer 35:1;O5, NAE 33:1;O4 298.7518 CerP 33:1;O2, LPC O-25:2, LPE O-28:2, CAR 26:0;O4 300.726 CAR 26:0;O4 300.72766 Cer 34:0;O6 300.74888 NAT 34:4 302.72456 CAR 30:2;O, Cer 37:3;O4, LPC O-25:0, LPE O-28:0 302.74585 ST 18:3;O4 303.13397 FA 20:4, ST 20:1;O2 303.23291 NAE 17:4 304.23643 CoA 10:0 306.07662 FA 13:3;O4 307.09887 ST 25:7;O7;GlcA 308.1022 FA 20:1 309.27987 HexCer 28:1;O3 314.72575 CerP 33:1;O4 316.72265 FA 22:6, ST 22:3;O2 327.2329 CAR 11:0, NAE 16:1;O2, SPB 18:2;O4, NAE 15:1, SPB 17:2;O2 328.23627 FA 22:4, ST 22:1;O2 331.26421 NAE 19:4 332.26761 ACer 65:0;O4 338.98868 ACer 64:3;O6, DGCC 54:2 342.96014 PIP3 31:7;O 345.08801 ST 18:5;O2;S 347.05899 ST 18:5;O5 351.09846 SHexCer 53:0;O4 358.9342 PC 56:0;O, PE 59:0;O, PS O-58:0, PT O-57:0 360.97079 SHexCer 29:6;O4 364.67041 SHexCer 28:5;O5 366.66734 PA O-66:1 368.66438 ACer 74:2;O2 369.02168 PIP 22:4 370.1349 CoA 24:2 370.139 TG 68:2;O2, TG O-68:3;O3 370.66149 ST 18:3;O6 371.12463 PIP2 43:12;O, PIP3 36:3;O 371.13825 NAT 11:2;O3 372.07128 CoA 23:0;O 372.14165 TG 70:5;O, TG O-70:6;O2 372.65896 HexCer 36:6;O4 373.74897 FA 20:2;O2, MG 17:2, MG O-17:3;O 375.23062 Hex2Cer 50:1;O5 376.94451 CoA 22:5;O4 380.10676 PIP 24:5 383.13034 LPA O-13:3 383.14283 TG 73:1, TG O-73:2;O 384.03243 CoA 24:5;O3 384.11419 SPBP 15:3;O3, M(IP)2C 28:5;O6 384.13794 ST 18:4;O7 385.1014 ST 19:3;O6 385.13892 ST 22:6;O;S 385.15018 CoA 27:7;O 386.13004 NAT 11:0;O3 386.14925 PIP 23:2;O, PIP3 41:6 387.15248 LPA O-16:2 391.22546 TG O-46:10;O3 395.27005 SQDG 33:2, FA 22:3;O2, MG 19:3, MG O-19:4;O, ST 22:0;O4 401.24593 TG 75:7;O2, TG O-75:8;O3 400.00646 LPE 10:2, SPBP 15:3;O4 400.13308 ST 22:6;O2;S 401.14081 NAT 10:0;O4, NAT 11:0;O4 402.14395 DGCC 68:10 402.99482 FA 22:2;O2, MG 19:2, MG O-19:3;O, SQDG 33:0 403.26167 MGDG 67:3 404.0144 TG 75:6;O3 406.01149 TG O-81:14, MGDG 68:4 408.01157 LPA 16:0, LPA O-16:1;O 409.23574 LPA O-18:3, FA 22:3;O3, MG 19:3;O, ST 22:0;O5 417.24091 CAR 13:1;O4 418.24422 CAR 15:1, NAE 20:2;O2, MGDG 41:9 418.27278 LPA O-18:2, FA 22:2;O3, MG 19:2;O 419.2564 SQDG 66:4, TG 82:18;O, TG O-82:19;O2 419.98814 CAR 13:0;O4, CAR 12:0;O2, NAE 17:1;O4, CAR 11:0;O2, NAE 16:1;O4 420.2601 TG 81:10;O2, TG O-81:11;O3 426.02205 LPA 18:1, LPA O-18:2;O 435.25176 LPC O-13:1, LPE O-16:1 436.28325 FA 22:1;O4, LPA 18:0, LPA O-18:1;O 437.26732 NAT 22:4 438.2704 DG 21:5, DG O-21:6;O, FA 24:6;O3, MG 21:6;O, ST 24:3;O5 439.2253 PI 68:1;O 442.01671 SHexCer 37:0;O6 442.77556 CAR 17:2, NAE 22:3;O2 444.2884 PA 49:6;O, PEth 47:6;O, PG O-46:7 444.31291 HexCer 45:6;O5 444.81424 CAR 17:1, NAE 22:2;O2, MGDG 45:9 446.30396 TG 87:19;O2, TG O-87:20;O3 448.00392 ST 19:3;O5;S 449.10053 LPC 13:0, LPC O-13:1;O, LPE 16:0, LPE O-16:1;O 452.27787 LPC O-15:2, LPE O-18:2 462.29869 ST 26:5;O5 463.22506 LPC O-15:1, LPE O-18:1, CAR 17:0;O, NAE 22:1;O3 464.31424 ST 20:3;O4;T 474.17231 LPC 15:1, LPC O-15:2;O, LPE 18:1, LPE O-18:2;O 478.29372 LPC 15:0, LPC O-15:1;O, LPE 18:0, LPE O-18:1;O, CAR 17:0;O2, NAE 22:1;O4 480.30924 PI O-45:7 481.31246 LPC 16:2;O, LPE 19:2;O, LPS O-18:2, LPT O-17:2, LPC O-15:3, LPE O-18:3 506.28865 ST 28:7;O6;S, ST 19:2;O5;GlcA, ST 19:3;O6;Hex 547.19255 PE 23:6, ST 27:4;O6;G, PC 21:6 552.27285 ACer 73:0;O4 558.06435 ST 18:1;O7;GlcA, ST 18:2;O8;Hex 577.2147 BMP 21:1;O, LPI O-18:2, PG 21:1;O 581.30909 LPI 18:0, LPI O-18:1;O, BMP 20:0, LPG 20:1;O, PG 20:0, PG O-20:1;O 599.31936 ST 30:0;O8;T, NAT 29:3;O4, ST 29:0;O6;T 648.37769 PIP2 56:2 650.3939 LPA 34:2;O, LPG O-31:3, PA 34:1, PA O-34:2;O 673.48028 HexCer 30:1;O4, CAR 28:1;O4, DGCC 25:0, HexCer 29:1;O2, CAR 27:1;O4 674.48315 LPG O-33:4, PA 36:2, PA O-36:3;O, LPA O-34:4 699.49591 HexCer 32:2;O4, CAR 29:2;O4, DGCC 26:1, HexCer 30:2;O2 700.49881 CerP 39:2;O3, LPC 31:2, LPC O-31:3;O 700.52703 LPG O-33:3, PA 36:1, PA O-36:2;O, PEth 34:1, LPA O-34:3 701.51139 HexCer 32:1;O4, CAR 29:1;O4, DGCC 26:0, HexCer 30:1;O2 702.51441 CerPE 38:1;O2 715.57465 CerP 39:2;O4, LPC 31:2;O, DGTA 30:1 716.52149 Cer 45:6;O2 716.57817 CerP 39:1;O4, LPC 31:1;O, PC 32:0 718.53787 CE 17:1;O3, DG 41:5 719.54139 CerP 41:5;O3, LPC 33:5 722.5112 PA 38:4, DG 39:5;O2, TG 39:4;O 723.49576 CerP 41:3;O3, LPC 33:3, LPC O-33:4;O, PE O-36:3 726.54329 TG 94:21;O3, CE 19:3;O2, DG O-43:8 727.54619 TG 94:20;O3, PC O-34:2, CerP 41:2;O3, LPC 33:2 728.55795 CerP 41:3;O4, LPC 33:3;O, PE 36:2 742.53681 CerP 41:2;O4, LPC 33:2;O, PE 36:1 744.55329 PA 40:7, PA O-40:8;O, PEth 38:7, DG 41:8;O2, TG 41:7;O, TG O-41:8;O2 745.48001 CL 74:0 745.5567 PE O-38:7 746.51154 PA 40:6, PA O-40:7;O, PEth 38:6, DG 41:7;O2, TG 41:6;O 747.49582 DG O-45:12 747.51381 HexCer 36:6;O4, CAR 34:6;O4, DGCC 31:5 748.49816 CerP 43:6;O3, PE O-38:6, PC O-36:6 748.52651 CerPE 37:3;O3, SM 34:3;O3 749.5019 CerP 43:5;O3, PE O-38:5 750.54202 PA 40:4, TG 41:4;O, DG 41:5;O2 751.52657 CE 21:5;O2, DG O-45:10 751.54605 CerP 43:3;O3, PE O-38:3, PC O-36:3 754.57434 PE 38:6, PE O-38:7;O, DGTA 34:6, PC 36:6 762.50556 CL 78:10 763.5097 CerP 43:6;O4, PE 38:5, LPC O-34:7 764.52187 CerP 43:5;O4, PE 38:4, DGTA 34:4, LPC O-34:6 766.53672 CE 21:5;O3, DG 45:9, CL 78:6 767.54068 PE O-40:8, CerP 39:0;O5, LPS O-33:0;O, PC O-38:8 772.52743 CerP 43:2;O4, PE 38:1, LPC O-34:3, PC 36:1 772.58508 PE O-40:7, PC O-38:7 774.54198 PG 36:1, PG O-36:2;O 775.54628 CerP 45:6;O3, PE O-40:6, PC O-38:6 776.55874 CerP 45:5;O3, PE O-40:5, PC O-38:5 778.57349 PE 40:7, PE O-40:8;O 788.52144 CerP 42:3;O6, PS 36:1, PT 35:1, LPC 33:3;O 788.54288 PA 43:6, MGDG 37:5, TG 41:5;O2 789.54657 PE 40:6, PC 38:6 790.53701 CL 82:10 791.54146 Hex2Cer 29:0;O2 792.54558 CerP 45:5;O4, PE 40:4, PC 38:4 794.56874 SHexCer 36:1;O2 806.54481 PS 40:6, PT 39:6, PC O-37:8;O, PE 40:7 834.52643 LPI 34:2;O, PG 35:2;O 835.53024 PC O-39:8, PE O-42:8 836.53326 SHexCer 38:1;O3 850.5697 IPC 36:2;O3 856.509 PI 36:4, PG 38:5;O 857.5157 PC O-41:11, PE O-44:11 858.51968 SHexCer 40:2;O2 860.59091 CerP 47:3;O4, PC 39:2, PE 42:2 862.60713 CerPE 42:2;O5, SM 39:2;O5 863.60935 CerP 47:4;O5, PC 39:3;O, PE 42:3;O 876.58473 SHexCer 41:1;O2 876.62184 HexCer 43:6;O6 878.60034 CerPE 42:2;O6, SM 39:2;O6 879.6045 PI 38:5, PI O-38:6;O 883.5329 PC O-43:12, PE O-46:12, Hex2Cer 34:5;O4 884.53578 PI 38:4, PG 40:5;O 885.54816 PC O-43:11, PE O-46:11, Hex2Cer 34:4;O4 886.55201 SHexCer 42:3;O2 886.60624 PG 42:5, PG O-42:6;O 887.55451 SM 41:4;O5 887.61068 SHexCer 42:2;O2 888.62284 TG 52:10;O3, CerPE 44:3;O5, SM 41:3;O5 889.62458 SHexCer 41:2;O3 890.60122 SHexCer 42:1;O2 890.63952 CerPE 43:2;O5, SM 40:2;O5 891.64216 CerP 48:3;O5, PC 40:2;O, PE 43:2;O, PS O-42:2, PT O-41:2 892.61773 CerPE 43:2;O6, SM 40:2;O6 893.62116 SHexCer 42:3;O3 902.60274 PC 41:3;O, PE 44:3;O, PS O-43:3, PT O-42:3 904.61757 CerPE 44:3;O6, SM 41:3;O6 905.62176 CerP 49:3;O5, PC 41:2;O, PE 44:2;O, PS O-43:2, PT O-42:2 906.63386 CerPE 44:2;O6, SM 41:2;O6 907.63685 TG 86:7 1324.21463 METABOLITES_END #END