#METABOLOMICS WORKBENCH DiegoC_20241001_130811 DATATRACK_ID:5254 STUDY_ID:ST003515 ANALYSIS_ID:AN005772 PROJECT_ID:PR002160 VERSION 1 CREATED_ON 10-10-2024 #PROJECT PR:PROJECT_TITLE Untargeted Metabolomics of 3xTg-AD Neurotoxic Astrocytes PR:PROJECT_SUMMARY Alzheimer's disease (AD) is the most common form of dementia, affecting PR:PROJECT_SUMMARY approximately 47M people worldwide. Histological features and genetic risk PR:PROJECT_SUMMARY factors, among other evidence, supported the amyloid hypothesis of the disease. PR:PROJECT_SUMMARY This neuronocentric paradigm is currently undergoing a shift, considering PR:PROJECT_SUMMARY evidence of the role of other cell types, such as microglia and astrocytes, in PR:PROJECT_SUMMARY disease progression. Previously, we described a particular astrocyte subtype PR:PROJECT_SUMMARY obtained from the 3xTg-AD murine model that displays neurotoxic properties in PR:PROJECT_SUMMARY vitro. We continue here our exploratory analysis through the lens of PR:PROJECT_SUMMARY metabolomics to identify potentially altered metabolites and biological PR:PROJECT_SUMMARY pathways. Cell extracts from neurotoxic and control astrocytes were compared PR:PROJECT_SUMMARY using HRMS-based metabolomics. Around 12% of metabolic features demonstrated PR:PROJECT_SUMMARY significant differences between neurotoxic and control astrocytes, including PR:PROJECT_SUMMARY alterations in the key metabolite glutamate. Consistent with our previous PR:PROJECT_SUMMARY transcriptomic study, the present results illustrate many homeostatic and PR:PROJECT_SUMMARY regulatory functions of metabolites, suggesting that neurotoxic 3xTg-AD PR:PROJECT_SUMMARY astrocytes exhibit alterations in the Krebs cycle as well as the prostaglandin PR:PROJECT_SUMMARY pathway. This is the first metabolomic study performed in 3xTg-AD neurotoxic PR:PROJECT_SUMMARY astrocytes. These results provide insight into metabolic alterations potentially PR:PROJECT_SUMMARY associated with neurotoxicity and pathology progression in the 3xTg-AD mouse PR:PROJECT_SUMMARY model and strengthen the therapeutic potential of astrocytes in AD. PR:INSTITUTE Instituto de Investigaciones Biológicas Clemente Estable (IIBCE) PR:DEPARTMENT NeuroquÃmica PR:LAST_NAME Carvalho PR:FIRST_NAME Diego PR:ADDRESS Isidoro de MarÃa 1614, Montevideo, Montevideo, 11800, Uruguay PR:EMAIL dicarez@fq.edu.uy PR:PHONE (+598) 2924 1879 PR:DOI http://dx.doi.org/10.21228/M8DR6G #STUDY ST:STUDY_TITLE Untargeted Metabolomics of 3xTg-AD Neurotoxic Astrocytes ST:STUDY_SUMMARY Alzheimer's disease (AD) is the most common form of dementia, affecting ST:STUDY_SUMMARY approximately 47M people worldwide. Histological features and genetic risk ST:STUDY_SUMMARY factors, among other evidence, supported the amyloid hypothesis of the disease. ST:STUDY_SUMMARY This neuronocentric paradigm is currently undergoing a shift, considering ST:STUDY_SUMMARY evidence of the role of other cell types, such as microglia and astrocytes, in ST:STUDY_SUMMARY disease progression. Previously, we described a particular astrocyte subtype ST:STUDY_SUMMARY obtained from the 3xTg-AD murine model that displays neurotoxic properties in ST:STUDY_SUMMARY vitro. We continue here our exploratory analysis through the lens of ST:STUDY_SUMMARY metabolomics to identify potentially altered metabolites and biological ST:STUDY_SUMMARY pathways. Cell extracts from neurotoxic and control astrocytes were compared ST:STUDY_SUMMARY using HRMS-based metabolomics. Around 12% of metabolic features demonstrated ST:STUDY_SUMMARY significant differences between neurotoxic and control astrocytes, including ST:STUDY_SUMMARY alterations in the key metabolite glutamate. Consistent with our previous ST:STUDY_SUMMARY transcriptomic study, the present results illustrate many homeostatic and ST:STUDY_SUMMARY regulatory functions of metabolites, suggesting that neurotoxic 3xTg-AD ST:STUDY_SUMMARY astrocytes exhibit alterations in the Krebs cycle as well as the prostaglandin ST:STUDY_SUMMARY pathway. This is the first metabolomic study performed in 3xTg-AD neurotoxic ST:STUDY_SUMMARY astrocytes. These results provide insight into metabolic alterations potentially ST:STUDY_SUMMARY associated with neurotoxicity and pathology progression in the 3xTg-AD mouse ST:STUDY_SUMMARY model and strengthen the therapeutic potential of astrocytes in AD. ST:INSTITUTE Instituto de Investigaciones Biológicas Clemente Estable (IIBCE) ST:LAST_NAME Carvalho ST:FIRST_NAME Diego ST:ADDRESS Isidoro de MarÃa 1614, Montevideo, Montevideo, 11800, Uruguay ST:EMAIL dicarez@fq.edu.uy ST:PHONE (+598) 2924 1879 ST:SUBMIT_DATE 2024-10-01 #SUBJECT SU:SUBJECT_TYPE Cultured cells SU:SUBJECT_SPECIES Mus musculus SU:TAXONOMY_ID 10090 SU:GENDER Female #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS 2A 2A_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_139.raw SUBJECT_SAMPLE_FACTORS 2A 2A_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_140.raw SUBJECT_SAMPLE_FACTORS 2A 2A_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_141.raw SUBJECT_SAMPLE_FACTORS 2A 2A_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_142.raw SUBJECT_SAMPLE_FACTORS 2A 2A_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_143.raw SUBJECT_SAMPLE_FACTORS 2A 2A_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_144.raw SUBJECT_SAMPLE_FACTORS 2B 2B_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_019.raw SUBJECT_SAMPLE_FACTORS 2B 2B_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_020.raw SUBJECT_SAMPLE_FACTORS 2B 2B_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_021.raw SUBJECT_SAMPLE_FACTORS 2B 2B_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_022.raw SUBJECT_SAMPLE_FACTORS 2B 2B_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_023.raw SUBJECT_SAMPLE_FACTORS 2B 2B_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_024.raw SUBJECT_SAMPLE_FACTORS 2C 2C_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_037.raw SUBJECT_SAMPLE_FACTORS 2C 2C_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_038.raw SUBJECT_SAMPLE_FACTORS 2C 2C_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_039.raw SUBJECT_SAMPLE_FACTORS 2C 2C_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; 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RAW_FILE_NAME=VT_190205_268.raw SUBJECT_SAMPLE_FACTORS 5B 5B_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_269.raw SUBJECT_SAMPLE_FACTORS 5B 5B_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_270.raw SUBJECT_SAMPLE_FACTORS 5C 5C_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_301.raw SUBJECT_SAMPLE_FACTORS 5C 5C_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_302.raw SUBJECT_SAMPLE_FACTORS 5C 5C_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_303.raw SUBJECT_SAMPLE_FACTORS 5C 5C_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Neonatal Column_Polarity=C18_Negative; 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RAW_FILE_NAME=VT_190205_327.raw SUBJECT_SAMPLE_FACTORS 4C 4C_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_328.raw SUBJECT_SAMPLE_FACTORS 4C 4C_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_329.raw SUBJECT_SAMPLE_FACTORS 4C 4C_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_330.raw SUBJECT_SAMPLE_FACTORS 7A 7A_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_073.raw SUBJECT_SAMPLE_FACTORS 7A 7A_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_074.raw SUBJECT_SAMPLE_FACTORS 7A 7A_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_075.raw SUBJECT_SAMPLE_FACTORS 7A 7A_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_076.raw SUBJECT_SAMPLE_FACTORS 7A 7A_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_077.raw SUBJECT_SAMPLE_FACTORS 7A 7A_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_078.raw SUBJECT_SAMPLE_FACTORS 7B 7B_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_067.raw SUBJECT_SAMPLE_FACTORS 7B 7B_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_068.raw SUBJECT_SAMPLE_FACTORS 7B 7B_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_069.raw SUBJECT_SAMPLE_FACTORS 7B 7B_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_070.raw SUBJECT_SAMPLE_FACTORS 7B 7B_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_071.raw SUBJECT_SAMPLE_FACTORS 7B 7B_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_072.raw SUBJECT_SAMPLE_FACTORS 7C 7C_1 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_043.raw SUBJECT_SAMPLE_FACTORS 7C 7C_2 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_044.raw SUBJECT_SAMPLE_FACTORS 7C 7C_3 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_045.raw SUBJECT_SAMPLE_FACTORS 7C 7C_4 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_046.raw SUBJECT_SAMPLE_FACTORS 7C 7C_5 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_047.raw SUBJECT_SAMPLE_FACTORS 7C 7C_6 Sample source:Cultured_astrocytes | Genotype:3xTg-AD | Condition:Old Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_048.raw SUBJECT_SAMPLE_FACTORS 1A 1A_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_091.raw SUBJECT_SAMPLE_FACTORS 1A 1A_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_092.raw SUBJECT_SAMPLE_FACTORS 1A 1A_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_093.raw SUBJECT_SAMPLE_FACTORS 1A 1A_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_094.raw SUBJECT_SAMPLE_FACTORS 1A 1A_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_095.raw SUBJECT_SAMPLE_FACTORS 1A 1A_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_096.raw SUBJECT_SAMPLE_FACTORS 1B 1B_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_211.raw SUBJECT_SAMPLE_FACTORS 1B 1B_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_212.raw SUBJECT_SAMPLE_FACTORS 1B 1B_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_213.raw SUBJECT_SAMPLE_FACTORS 1B 1B_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_214.raw SUBJECT_SAMPLE_FACTORS 1B 1B_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_215.raw SUBJECT_SAMPLE_FACTORS 1B 1B_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_216.raw SUBJECT_SAMPLE_FACTORS 1C 1C_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_055.raw SUBJECT_SAMPLE_FACTORS 1C 1C_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_056.raw SUBJECT_SAMPLE_FACTORS 1C 1C_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_057.raw SUBJECT_SAMPLE_FACTORS 1C 1C_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_058.raw SUBJECT_SAMPLE_FACTORS 1C 1C_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_059.raw SUBJECT_SAMPLE_FACTORS 1C 1C_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_060.raw SUBJECT_SAMPLE_FACTORS 6A 6A_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_253.raw SUBJECT_SAMPLE_FACTORS 6A 6A_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_254.raw SUBJECT_SAMPLE_FACTORS 6A 6A_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_255.raw SUBJECT_SAMPLE_FACTORS 6A 6A_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_256.raw SUBJECT_SAMPLE_FACTORS 6A 6A_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_257.raw SUBJECT_SAMPLE_FACTORS 6A 6A_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_258.raw SUBJECT_SAMPLE_FACTORS 6B 6B_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_115.raw SUBJECT_SAMPLE_FACTORS 6B 6B_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_116.raw SUBJECT_SAMPLE_FACTORS 6B 6B_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_117.raw SUBJECT_SAMPLE_FACTORS 6B 6B_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_118.raw SUBJECT_SAMPLE_FACTORS 6B 6B_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_119.raw SUBJECT_SAMPLE_FACTORS 6B 6B_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_120.raw SUBJECT_SAMPLE_FACTORS 6C 6C_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_085.raw SUBJECT_SAMPLE_FACTORS 6C 6C_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_086.raw SUBJECT_SAMPLE_FACTORS 6C 6C_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_087.raw SUBJECT_SAMPLE_FACTORS 6C 6C_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_088.raw SUBJECT_SAMPLE_FACTORS 6C 6C_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_089.raw SUBJECT_SAMPLE_FACTORS 6C 6C_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_090.raw SUBJECT_SAMPLE_FACTORS 8A 8A_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_205.raw SUBJECT_SAMPLE_FACTORS 8A 8A_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_206.raw SUBJECT_SAMPLE_FACTORS 8A 8A_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_207.raw SUBJECT_SAMPLE_FACTORS 8A 8A_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_208.raw SUBJECT_SAMPLE_FACTORS 8A 8A_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_209.raw SUBJECT_SAMPLE_FACTORS 8A 8A_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_210.raw SUBJECT_SAMPLE_FACTORS 8B 8B_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_109.raw SUBJECT_SAMPLE_FACTORS 8B 8B_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_110.raw SUBJECT_SAMPLE_FACTORS 8B 8B_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_111.raw SUBJECT_SAMPLE_FACTORS 8B 8B_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_112.raw SUBJECT_SAMPLE_FACTORS 8B 8B_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_113.raw SUBJECT_SAMPLE_FACTORS 8B 8B_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_114.raw SUBJECT_SAMPLE_FACTORS 8C 8C_1 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_049.raw SUBJECT_SAMPLE_FACTORS 8C 8C_2 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_050.raw SUBJECT_SAMPLE_FACTORS 8C 8C_3 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_051.raw SUBJECT_SAMPLE_FACTORS 8C 8C_4 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_052.raw SUBJECT_SAMPLE_FACTORS 8C 8C_5 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_053.raw SUBJECT_SAMPLE_FACTORS 8C 8C_6 Sample source:Cultured_astrocytes | Genotype:Wild-type | Condition:Neonatal Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_054.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_1 q3June2014_1a_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_007.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_2 q3June2014_1a_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_008.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_3 q3June2014_1a_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_009.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_4 q3June2014_1a_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_010.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_5 q3June2014_1a_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_011.raw SUBJECT_SAMPLE_FACTORS q3June2014_1a_6 q3June2014_1a_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_012.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_1 q3June2014_1b_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_013.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_2 q3June2014_1b_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_014.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_3 q3June2014_1b_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_015.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_4 q3June2014_1b_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_016.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_5 q3June2014_1b_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_017.raw SUBJECT_SAMPLE_FACTORS q3June2014_1b_6 q3June2014_1b_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_018.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_1 q3June2014_1c_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_193.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_2 q3June2014_1c_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_194.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_3 q3June2014_1c_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_195.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_4 q3June2014_1c_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_196.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_5 q3June2014_1c_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_197.raw SUBJECT_SAMPLE_FACTORS q3June2014_1c_6 q3June2014_1c_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_198.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_1 q3June2014_1d_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_199.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_2 q3June2014_1d_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_200.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_3 q3June2014_1d_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_201.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_4 q3June2014_1d_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_202.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_5 q3June2014_1d_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_203.raw SUBJECT_SAMPLE_FACTORS q3June2014_1d_6 q3June2014_1d_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_204.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_1 q3June2014_1e_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_373.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_2 q3June2014_1e_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_374.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_3 q3June2014_1e_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_375.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_4 q3June2014_1e_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_376.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_5 q3June2014_1e_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_377.raw SUBJECT_SAMPLE_FACTORS q3June2014_1e_6 q3June2014_1e_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_378.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_1 q3June2014_1f_1 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_379.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_2 q3June2014_1f_2 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_380.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_3 q3June2014_1f_3 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_381.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_4 q3June2014_1f_4 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_382.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_5 q3June2014_1f_5 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_383.raw SUBJECT_SAMPLE_FACTORS q3June2014_1f_6 q3June2014_1f_6 Sample source:Pooled_human_plasma | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_384.raw SUBJECT_SAMPLE_FACTORS nist_1 nist_1 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_001.raw SUBJECT_SAMPLE_FACTORS nist_2 nist_2 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_002.raw SUBJECT_SAMPLE_FACTORS nist2_1 nist2_1 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_385.raw SUBJECT_SAMPLE_FACTORS nist2_2 nist2_2 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_386.raw SUBJECT_SAMPLE_FACTORS nist2_3 nist2_3 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_387.raw SUBJECT_SAMPLE_FACTORS nist2_4 nist2_4 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_388.raw SUBJECT_SAMPLE_FACTORS nist2_5 nist2_5 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_389.raw SUBJECT_SAMPLE_FACTORS nist2_6 nist2_6 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_390.raw SUBJECT_SAMPLE_FACTORS nist_3 nist_3 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_003.raw SUBJECT_SAMPLE_FACTORS nist_4 nist_4 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_004.raw SUBJECT_SAMPLE_FACTORS nist_5 nist_5 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=HILIC_Positive; RAW_FILE_NAME=VT_190205_005.raw SUBJECT_SAMPLE_FACTORS nist_6 nist_6 Sample source:Reference_standards | Genotype:- | Condition:Quality_control Column_Polarity=C18_Negative; RAW_FILE_NAME=VT_190205_006.raw #COLLECTION CO:COLLECTION_SUMMARY Neurotoxic astrocyte cultures were prepared from the cerebral cortex and CO:COLLECTION_SUMMARY hippocampus of 9- to 10-month-old 3xTg-AD female mice according to previously CO:COLLECTION_SUMMARY described methods [2]. Female animals were used since sex differences in the CO:COLLECTION_SUMMARY development of the pathology have been described in this AD murine model, with a CO:COLLECTION_SUMMARY greater expression of transgenes in females [3,4]. Astrocytes isolated from 9- CO:COLLECTION_SUMMARY to 10-month-old non-Tg mice were not used as controls in our studies due to the CO:COLLECTION_SUMMARY low yield achieved [2]. Instead, astrocytic cultures derived from neonatal CO:COLLECTION_SUMMARY non-Tg mice as well as astrocytes from neonatal 3xTg-AD mice were used as CO:COLLECTION_SUMMARY controls. These non-toxic controls allowed us to assess whether astrocytes from CO:COLLECTION_SUMMARY this mice model of AD exhibit alterations when isolated before the disease CO:COLLECTION_SUMMARY onset. Nontoxic control astrocyte cultures were derived from the cerebral cortex CO:COLLECTION_SUMMARY and hippocampus of neonatal (postnatal day 0â2) 3xTg-AD and non-Tg (C57BL/6J) CO:COLLECTION_SUMMARY mice following the methods described by Cassina et al. with minor modifications CO:COLLECTION_SUMMARY [2,5]. All cell cultures were amplified and maintained at 37°C in a humidified CO:COLLECTION_SUMMARY incubator with 5% CO2. References [2] P. Diaz-Amarilla, F. Arredondo, R. CO:COLLECTION_SUMMARY Dapueto, V. Boix, D. Carvalho, M.D. Santi, E. Vasilskis, R. Mesquita-Ribeiro, F. CO:COLLECTION_SUMMARY Dajas-Bailador, J.A. Abin-Carriquiry, H. Engler, E. Savio, Isolation and CO:COLLECTION_SUMMARY characterization of neurotoxic astrocytes derived from old triple transgenic CO:COLLECTION_SUMMARY Alzheimerâs disease mice, Neurochem. Int. 159 (2022). CO:COLLECTION_SUMMARY https://doi.org/10.1016/j.neuint.2022.105403. [3] J.C. Carroll, E.R. Rosario, S. CO:COLLECTION_SUMMARY Kreimer, A. Villamagna, E. Gentzschein, F.Z. Stanczyk, C.J. Pike, Sex CO:COLLECTION_SUMMARY differences in β-amyloid accumulation in 3xTg-AD mice: Role of neonatal sex CO:COLLECTION_SUMMARY steroid hormone exposure, Brain Res. 1366 (2010) 233â245. CO:COLLECTION_SUMMARY https://doi.org/10.1016/J.BRAINRES.2010.10.009. [4] L.K. Clinton, L.M. Billings, CO:COLLECTION_SUMMARY K.N. Green, A. Caccamo, J. Ngo, S. Oddo, J.L. McGaugh, F.M. LaFerla, CO:COLLECTION_SUMMARY Age-dependent sexual dimorphism in cognition and stress response in the 3xTg-AD CO:COLLECTION_SUMMARY mice, Neurobiol. Dis. 28 (2007) 76â82. CO:COLLECTION_SUMMARY https://doi.org/10.1016/J.NBD.2007.06.013. [5] P. Cassina, H. Peluffo, M. Pehar, CO:COLLECTION_SUMMARY L. Martinez-Palma, A. Ressia, J.S. Beckman, A.G. Estévez, L. Barbeito, CO:COLLECTION_SUMMARY Peroxynitrite triggers a phenotypic transformation in spinal cord astrocytes CO:COLLECTION_SUMMARY that induces motor neuron apoptosis, J. Neurosci. Res. 67 (2002) 21â29. CO:COLLECTION_SUMMARY https://doi.org/10.1002/jnr.10107. CO:SAMPLE_TYPE Astrocytes #TREATMENT TR:TREATMENT_SUMMARY No treatment #SAMPLEPREP SP:SAMPLEPREP_SUMMARY Extraction of metabolites from cultured cells was performed based on previous SP:SAMPLEPREP_SUMMARY work by Go et al. (2015), Liu et al. (2019), as well as Sapcariu et al. (2014) SP:SAMPLEPREP_SUMMARY [6-8]. One sample consisted of metabolites extracted and pooled from 3 wells of SP:SAMPLEPREP_SUMMARY a 6-well plate, pooling a total of 9 samples per condition. Briefly, confluent SP:SAMPLEPREP_SUMMARY cells were washed with 0.9% (m/v) NaCl at room temperature, followed by the SP:SAMPLEPREP_SUMMARY addition of 300 µL of an ice-cold HPLC-grade acetonitrile-water solution SP:SAMPLEPREP_SUMMARY mixture (1:2) per well. This allowed us to scrape cells, precipitate proteins, SP:SAMPLEPREP_SUMMARY and extract metabolites. The extracts were kept at 4 °C for 30 min. and SP:SAMPLEPREP_SUMMARY centrifuged at 16,100 g for 10 min to remove the protein and any remaining SP:SAMPLEPREP_SUMMARY insoluble fraction. Supernatants were transferred to screw-cap vials and dried SP:SAMPLEPREP_SUMMARY in a fast vacuum centrifuge (overnight, 35°C). #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Solvent A: Water; Solvent B: Acetonitrile; Solvent C: 2% formic acid (v/v) in CH:CHROMATOGRAPHY_SUMMARY water CH:INSTRUMENT_NAME Thermo Dionex Ultimate 3000 CH:COLUMN_NAME Thermo Accucore HILIC (50 x 2.1mm,2.6um) CH:COLUMN_TEMPERATURE 60 CH:FLOW_GRADIENT Mobile phase conditions consisted of 22.5% A, 75% B, 2.5% C which was held for CH:FLOW_GRADIENT 1.5 min, with a linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, and held for CH:FLOW_GRADIENT 1 min CH:FLOW_RATE Flow rate of the HILIC column was maintained at 0.35 mL/min until 1.5 min, CH:FLOW_RATE increased to 0.4 mL/min at 4 min and held for 1 min, resulting in a total CH:FLOW_RATE analytical run time of 5 min CH:SOLVENT_A 100% water CH:SOLVENT_B 100% acetonitrile CH:CHROMATOGRAPHY_TYPE HILIC CH:SOLVENT_C 98% water/2% formic acid #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:MS_COMMENTS MS acquisition Comments: Collected mass spectral data were analyzed for features MS:MS_COMMENTS using Xcaliber (ThermoFisher; Waltham MA). Ultra-high resolution mass MS:MS_COMMENTS spectrometry operating at 60,000 and 120,000 resolution has previously been MS:MS_COMMENTS shown to provide effective metabolite quantification in biological extracts [11] MS:MS_COMMENTS and the use of complementary chromatography and ionization phases have been MS:MS_COMMENTS shown to improve the detection of endogenous and exogenous chemicals. Data MS:MS_COMMENTS processing Comments: Raw mass spectral data files were converted to computable MS:MS_COMMENTS document format (CDF) using Xcalibur file conversion software (Thermo Fisher MS:MS_COMMENTS Scientific, San Diego, CA) for further processing. Software/procedures used for MS:MS_COMMENTS feature assignments: Data were processed for peak extraction, noise filtering, MS:MS_COMMENTS m/z and retention time alignment, and quantification of ion intensities using MS:MS_COMMENTS apLCMS [29] with enhanced data extraction using xMSanalyzer [30]. xMSanalyzer MS:MS_COMMENTS improves feature detection through systematic data re-extraction, statistical MS:MS_COMMENTS filtering and fusion [30]. Metabolite feature peak intensity values ââwere MS:MS_COMMENTS summarized by the median in triplicate (technical replicates) and subjected to MS:MS_COMMENTS quality assessment. The samples were filtered considering an overall Pearson MS:MS_COMMENTS correlation of technical replicates of (r) > 0.70 and a cut-off coefficient MS:MS_COMMENTS of variation of 75%. Considering the number of missing values, one sample MS:MS_COMMENTS outlier was further removed from the C18 and HILIC cell extraction feature MS:MS_COMMENTS tables. Attached is the xMSAnalyzer table of the detected peak intensities. As MS:MS_COMMENTS stated above, the values âârepresent the median of three technical MS:MS_COMMENTS replicates evaluated per original sample. For example, samples labeled 2A_1, MS:MS_COMMENTS 2A_3 and 2A_5 in the design table are summarized under label 2A in the features MS:MS_COMMENTS table. For further details on the procedure, please refer to the xMSAnalyzer MS:MS_COMMENTS publication. Uppal, K., Soltow, Q.A., Strobel, F.H. et al. xMSanalyzer: MS:MS_COMMENTS automated pipeline for improved feature detection and downstream analysis of MS:MS_COMMENTS large-scale, non-targeted metabolomics data. BMC Bioinformatics 14, 15 (2013). MS:MS_COMMENTS https://doi.org/10.1186/1471-2105-14-15 MS:ION_MODE POSITIVE MS:MS_RESULTS_FILE ST003515_AN005772_Results.txt UNITS:m/z Has m/z:Yes Has RT:Yes RT units:Seconds #END