#METABOLOMICS WORKBENCH stephensondj_20240227_094013 DATATRACK_ID:4676 STUDY_ID:ST003108 ANALYSIS_ID:AN005087 PROJECT_ID:PR001931
VERSION             	1
CREATED_ON             	February 27, 2024, 10:57 am
#PROJECT
PR:PROJECT_TITLE                 	Complete absence of GLUT1 does not impair human terminal erythroid
PR:PROJECT_TITLE                 	differentiation
PR:PROJECT_SUMMARY               	The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within
PR:PROJECT_SUMMARY               	the erythrocyte membrane and is required for glucose and dehydroascorbic acid
PR:PROJECT_SUMMARY               	(Vitamin C precursor) transport. It is widely recognized as a key protein for
PR:PROJECT_SUMMARY               	red cell structure, function, and metabolism. Previous reports highlighted the
PR:PROJECT_SUMMARY               	importance of GLUT1 activity within these uniquely glycolysis-dependent cells,
PR:PROJECT_SUMMARY               	in particular for increasing antioxidant capacity needed to avoid irreversible
PR:PROJECT_SUMMARY               	damage from oxidative stress in humans. However, studies of glucose transporter
PR:PROJECT_SUMMARY               	roles in erythroid cells are complicated by species-specific differences between
PR:PROJECT_SUMMARY               	humans and mice. Here, using CRISPRmediated gene editing of immortalized
PR:PROJECT_SUMMARY               	erythroblasts and adult CD34+ hematopoietic progenitor cells, we generate
PR:PROJECT_SUMMARY               	committed human erythroid cells completely deficient in expression of GLUT1. We
PR:PROJECT_SUMMARY               	show that absence of GLUT1 does not impede human erythroblast proliferation,
PR:PROJECT_SUMMARY               	differentiation, or enucleation. This work demonstrates for the first-time
PR:PROJECT_SUMMARY               	generation of enucleated human reticulocytes lacking GLUT1. The GLUT1-deficient
PR:PROJECT_SUMMARY               	reticulocytes possess no tangible alterations to membrane composition or
PR:PROJECT_SUMMARY               	deformability in reticulocytes. Metabolomic analyses of GLUT1-deficient
PR:PROJECT_SUMMARY               	reticulocytes reveal hallmarks of reduced glucose import, downregulated
PR:PROJECT_SUMMARY               	metabolic processes and upregulated AMPK-signalling, alongside alterations in
PR:PROJECT_SUMMARY               	antioxidant metabolism, resulting in increased osmotic fragility and metabolic
PR:PROJECT_SUMMARY               	shifts indicative of higher oxidant stress. Despite detectable metabolic changes
PR:PROJECT_SUMMARY               	in GLUT1 deficient reticulocytes, the absence of developmental phenotype,
PR:PROJECT_SUMMARY               	detectable proteomic compensation or impaired deformability comprehensively
PR:PROJECT_SUMMARY               	alters our understanding of the role of GLUT1 in red blood cell structure,
PR:PROJECT_SUMMARY               	function and metabolism. It also provides cell biological evidence supporting
PR:PROJECT_SUMMARY               	clinical consensus that reduced GLUT1 expression does not cause anaemia in GLUT1
PR:PROJECT_SUMMARY               	deficiency syndrome.
PR:INSTITUTE                     	University of Colorado
PR:LAST_NAME                     	Stephenson
PR:FIRST_NAME                    	Daniel
PR:ADDRESS                       	Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA
PR:EMAIL                         	daniel.stephenson@cuanschutz.edu
PR:PHONE                         	303-724-3339
#STUDY
ST:STUDY_TITLE                   	Complete absence of GLUT1 does not impair human terminal erythroid
ST:STUDY_TITLE                   	differentiation
ST:STUDY_SUMMARY                 	The Glucose transporter 1 (GLUT1) is one of the most abundant proteins within
ST:STUDY_SUMMARY                 	the erythrocyte membrane and is required for glucose and dehydroascorbic acid
ST:STUDY_SUMMARY                 	(Vitamin C precursor) transport. It is widely recognized as a key protein for
ST:STUDY_SUMMARY                 	red cell structure, function, and metabolism. Previous reports highlighted the
ST:STUDY_SUMMARY                 	importance of GLUT1 activity within these uniquely glycolysis-dependent cells,
ST:STUDY_SUMMARY                 	in particular for increasing antioxidant capacity needed to avoid irreversible
ST:STUDY_SUMMARY                 	damage from oxidative stress in humans. However, studies of glucose transporter
ST:STUDY_SUMMARY                 	roles in erythroid cells are complicated by species-specific differences between
ST:STUDY_SUMMARY                 	humans and mice. Here, using CRISPRmediated gene editing of immortalized
ST:STUDY_SUMMARY                 	erythroblasts and adult CD34+ hematopoietic progenitor cells, we generate
ST:STUDY_SUMMARY                 	committed human erythroid cells completely deficient in expression of GLUT1. We
ST:STUDY_SUMMARY                 	show that absence of GLUT1 does not impede human erythroblast proliferation,
ST:STUDY_SUMMARY                 	differentiation, or enucleation. This work demonstrates for the first-time
ST:STUDY_SUMMARY                 	generation of enucleated human reticulocytes lacking GLUT1. The GLUT1-deficient
ST:STUDY_SUMMARY                 	reticulocytes possess no tangible alterations to membrane composition or
ST:STUDY_SUMMARY                 	deformability in reticulocytes. Metabolomic analyses of GLUT1-deficient
ST:STUDY_SUMMARY                 	reticulocytes reveal hallmarks of reduced glucose import, downregulated
ST:STUDY_SUMMARY                 	metabolic processes and upregulated AMPK-signalling, alongside alterations in
ST:STUDY_SUMMARY                 	antioxidant metabolism, resulting in increased osmotic fragility and metabolic
ST:STUDY_SUMMARY                 	shifts indicative of higher oxidant stress. Despite detectable metabolic changes
ST:STUDY_SUMMARY                 	in GLUT1 deficient reticulocytes, the absence of developmental phenotype,
ST:STUDY_SUMMARY                 	detectable proteomic compensation or impaired deformability comprehensively
ST:STUDY_SUMMARY                 	alters our understanding of the role of GLUT1 in red blood cell structure,
ST:STUDY_SUMMARY                 	function and metabolism. It also provides cell biological evidence supporting
ST:STUDY_SUMMARY                 	clinical consensus that reduced GLUT1 expression does not cause anaemia in GLUT1
ST:STUDY_SUMMARY                 	deficiency syndrome.
ST:INSTITUTE                     	University of Colorado
ST:LAST_NAME                     	Stephenson
ST:FIRST_NAME                    	Daniel
ST:ADDRESS                       	Research 1 South L18-1303 12801 E. 17th Ave., Aurora, Colorado, 80045, USA
ST:EMAIL                         	daniel.stephenson@cuanschutz.edu
ST:PHONE                         	303-724-3339
#SUBJECT
SU:SUBJECT_TYPE                  	Cultured cells
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#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           	Donor 1	DS1-095-001+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-001+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-002+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-002+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-003+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-003+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-004+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-004+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-005+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-005+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-006+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-006+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-007+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-007+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-008+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-008+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-009+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-009+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-010+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-010+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-011+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-011+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-012+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-012+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-013+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-013+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-014+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-014+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-015+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-015+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-016+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-016+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-017+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-017+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-018+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-018+.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-001-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-001-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-002-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-002-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-003-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-003-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-004-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-004-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-005-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-005-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-006-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-006-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-007-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-007-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-008-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-008-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-009-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-009-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-010-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-010-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-011-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-011-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-012-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-012-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-013-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-013-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-014-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-014-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-015-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-015-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-016-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-016-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-017-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-017-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-018-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-018-.mzXML; Pellet size=10 Million; Analysis=Metabolomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-001L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-001L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-002L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-002L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-003L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-003L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-004L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-004L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-005L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-005L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-006L+	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-006L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-007L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-007L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-008L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-008L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-009L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-009L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-010L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-010L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-011L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-011L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-012L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-012L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-013L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-013L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-014L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-014L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-015L+	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-015L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-016L+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-016L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-017L+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-017L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-018L+	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-018L+.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Positive
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-001L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-001L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 1	DS1-095-002L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-002L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-003L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-003L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 4	DS1-095-004L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-004L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-005L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-005L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor 5	DS1-095-006L-	Sample source:cultured reticulocytes | Assay:04CF29	RAW_FILE_NAME(Raw File Name)=DS1-095-006L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-007L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-007L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-008L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-008L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-009L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-009L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-010L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-010L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-011L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-011L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-012L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-012L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-013L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-013L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-014L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-014L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-015L-	Sample source:cultured reticulocytes | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-015L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor A	DS1-095-016L-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-016L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor C	DS1-095-017L-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-017L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
SUBJECT_SAMPLE_FACTORS           	Donor D	DS1-095-018L-	Sample source:Red blood cells | Assay:01CF19	RAW_FILE_NAME(Raw File Name)=DS1-095-018L-.RAW; Pellet size=10 Million; Analysis=Lipidomics; Polarity=Negative
#COLLECTION
CO:COLLECTION_SUMMARY            	For flow cytometry, cells were fixed (1% paraformaldehyde, 0.0075%
CO:COLLECTION_SUMMARY            	glutaraldehyde) to prevent antibody agglutination, except for assays using
CO:COLLECTION_SUMMARY            	GLUT1.RBD. Samples were labelled with primary antibodies in PBSAG (PBS with 1%
CO:COLLECTION_SUMMARY            	(w/v) Glucose and 0.5%(w/v) |Bovine Serum Albumin (BSA)) supplemented with extra
CO:COLLECTION_SUMMARY            	1%(w/v) BSA for 30min, in the dark. METAFORA’s GLUT1.RBD was incubated at
CO:COLLECTION_SUMMARY            	37°C and all remaining antibodies incubated at 4°C. Cells were washed 2x PBSAG
CO:COLLECTION_SUMMARY            	and, if required, incubated with appropriate secondary antibody under the same
CO:COLLECTION_SUMMARY            	conditions as described for primary. Cells were washed 2x with PBSAG and
CO:COLLECTION_SUMMARY            	analysed on a Miltenyi MACSQuant 10 flow cytometer. Data was analysed using
CO:COLLECTION_SUMMARY            	FlowJo v10.7 (FlowJo). Reticulocytes were identified by gating on the
CO:COLLECTION_SUMMARY            	Hoechst-negative population. 4 Cells were sorted using a BD Influx Cell Sorter
CO:COLLECTION_SUMMARY            	(BD Biosciences). BEL-A CRISPR edited populations were single cell sorted based
CO:COLLECTION_SUMMARY            	on viability (DRAQ7 negativity). Primary GLUT1 KO cultures were sorted on days 6
CO:COLLECTION_SUMMARY            	or 7 of differentiation using DRAQ7 and the eGFP-fused GLUT1.RBD to purify the
CO:COLLECTION_SUMMARY            	negative population with a gate based on non-targeting guide control cells.
CO:SAMPLE_TYPE                   	Cultured cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Filtered reticulocytes (1-2 x105/well) were incubated in decreasing NaCl
TR:TREATMENT_SUMMARY             	concentrations (0.9–0%) for 10min at 37°C. Lysis was stopped by adding 4x
TR:TREATMENT_SUMMARY             	volume of PBSAG. Live cells, considered as having a normal FSC/SSC profile as
TR:TREATMENT_SUMMARY             	defined by the 0.9% NaCl control, were counted by flow cytometry using the
TR:TREATMENT_SUMMARY             	MACSQuant10
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Extraction of metabolites and lipids from red cell pellets was as follows: 36
SP:SAMPLEPREP_SUMMARY            	µL of cold MeOH:MeCN:H2O (5:3:2, v:v:v) was added to all pellets (pellet size
SP:SAMPLEPREP_SUMMARY            	pre-determined to be 10 million prior to extraction). Samples were vortexed at
SP:SAMPLEPREP_SUMMARY            	4°C for 30 minutes. Insoluble material was pelleted by centrifugation (18000g,
SP:SAMPLEPREP_SUMMARY            	10 min) and supernatants were isolated for analysis by UHPLC-MS for metabolomics
SP:SAMPLEPREP_SUMMARY            	and lipidomics.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Metabolomics Positive
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	0.1% Formic Acid in Water
CH:SOLVENT_B                     	0.1% Formic Acid in ACN
CH:FLOW_GRADIENT                 	0 min - 0.45 ml/min - 5% B, 0.5 min - 0.45ml/min - 5% B, 1.1 min - 0.45ml/min -
CH:FLOW_GRADIENT                 	95% B, 2.75 min - 0.45ml/min - 95% B, 3 min - 0.45ml/min - 5% B, 5min -
CH:FLOW_GRADIENT                 	0.45ml/min - 5%B
CH:FLOW_RATE                     	0.45 ml/min
CH:COLUMN_TEMPERATURE            	45
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	El-Maven used for data analysis
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	area
MS_METABOLITE_DATA_START
Samples	DS1-095-001+	DS1-095-002+	DS1-095-003+	DS1-095-004+	DS1-095-005+	DS1-095-006+	DS1-095-007+	DS1-095-008+	DS1-095-009+	DS1-095-010+	DS1-095-011+	DS1-095-012+	DS1-095-013+	DS1-095-014+	DS1-095-015+	DS1-095-016+	DS1-095-017+	DS1-095-018+
Factors	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:04CF29	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:cultured reticulocytes | Assay:01CF19	Sample source:Red blood cells | Assay:01CF19	Sample source:Red blood cells | Assay:01CF19	Sample source:Red blood cells | Assay:01CF19
L-alanine	4.73E+06	4.35E+06	2.75E+06	3.20E+06	4.96E+06	4.77E+06	3.88E+06	3.76E+06	3.26E+06	7.96E+06	9.62E+06	4.75E+06	5.32E+06	3.16E+06	6.33E+06	1.04E+06	9.02E+05	1.14E+06
L-arginine	8.78E+04	9.29E+04	1.00E+05	6.25E+04	2.20E+05	2.55E+05	4.54E+04	6.00E+04	1.09E+05	1.02E+05	1.79E+05	2.26E+05	5.65E+04	4.77E+04	5.12E+05	9.18E+04	5.36E+04	8.62E+04
L-asparagine	3.21E+05	2.12E+05	1.62E+05	1.32E+05	2.00E+05	1.63E+05	1.56E+05	1.34E+05	1.40E+05	3.58E+05	4.24E+05	1.47E+05	1.94E+05	1.37E+05	2.04E+05	4.03E+04	3.14E+04	8.58E+04
L-cysteine	2.36E+04	1.91E+04	2.04E+04	2.31E+04	2.55E+04	1.56E+04	2.07E+04	2.41E+04	2.14E+04	3.03E+04	3.52E+04	4.81E+04	3.68E+04	2.58E+04	2.96E+04	1.91E+04	2.57E+04	1.67E+04
glycine	8.29E+05	6.56E+05	5.84E+05	5.64E+05	9.29E+05	6.01E+05	1.02E+06	8.83E+05	6.85E+05	1.65E+06	2.25E+06	8.52E+05	1.53E+06	9.25E+05	1.78E+06	1.80E+05	1.67E+05	2.83E+05
L-histidine	6.87E+05	4.95E+05	2.23E+05	2.16E+05	7.71E+05	6.56E+05	7.96E+05	7.31E+05	3.98E+05	2.14E+06	1.76E+06	4.31E+05	3.59E+05	2.18E+05	3.48E+05	5.33E+05	3.97E+05	5.96E+05
L-leucine/isoluecine	4.42E+05	4.54E+05	4.35E+05	4.15E+05	3.81E+05	3.85E+05	5.79E+05	4.74E+05	8.98E+05	5.82E+05	6.18E+05	7.47E+05	7.76E+05	8.99E+05	1.06E+06	3.59E+05	3.26E+05	3.32E+05
L-lysine	6.50E+05	4.05E+05	3.25E+05	2.31E+05	6.36E+05	5.07E+05	7.62E+05	7.59E+05	5.47E+05	1.70E+06	1.80E+06	5.29E+05	6.52E+05	5.65E+05	6.49E+05	4.56E+05	3.23E+05	5.00E+05
L-methionine	5.70E+04	4.82E+04	4.42E+04	3.56E+04	5.07E+04	5.09E+04	7.09E+04	4.62E+04	6.32E+04	9.96E+04	6.34E+04	3.74E+04	8.59E+04	4.92E+04	8.53E+04	1.05E+04	1.61E+04	4.94E+04
L-phenylalanine	2.71E+05	1.87E+05	2.35E+05	2.58E+05	1.76E+05	2.38E+05	2.75E+05	2.63E+05	4.25E+05	3.37E+05	2.79E+05	3.98E+05	3.29E+05	4.38E+05	4.96E+05	1.63E+05	1.75E+05	1.40E+05
L-proline	5.52E+06	6.53E+06	3.12E+06	4.74E+06	5.22E+06	5.82E+06	4.54E+06	3.45E+06	5.00E+06	1.06E+07	1.29E+07	8.09E+06	6.94E+06	3.85E+06	1.30E+07	2.72E+06	2.23E+06	3.88E+06
L-serine	3.49E+05	2.81E+05	1.59E+05	1.32E+05	3.15E+05	1.93E+05	4.81E+05	2.62E+05	3.59E+05	7.11E+05	6.93E+05	2.59E+05	7.04E+05	2.28E+05	7.23E+05	7.90E+04	6.85E+04	1.20E+05
L-threonine	6.99E+05	8.47E+05	3.28E+05	4.84E+05	7.30E+05	7.77E+05	7.99E+05	5.25E+05	7.12E+05	1.64E+06	1.67E+06	1.11E+06	1.16E+06	5.76E+05	1.78E+06	1.28E+05	1.17E+05	1.61E+05
L-tryptophan	5.51E+05	9.18E+05	5.66E+05	1.27E+06	9.15E+05	1.45E+06	8.88E+05	5.24E+05	6.06E+05	2.28E+06	1.43E+06	1.07E+06	7.83E+05	5.48E+05	1.12E+06	7.65E+05	8.22E+05	5.58E+05
L-tyrosine	1.52E+05	2.70E+05	1.55E+05	2.40E+05	3.05E+05	4.54E+05	2.30E+05	1.78E+05	1.81E+05	8.65E+05	4.28E+05	1.43E+05	2.38E+05	1.24E+05	3.32E+05	2.02E+05	1.76E+05	1.94E+05
L-valine	5.73E+06	4.97E+06	2.50E+06	2.15E+06	4.76E+06	4.56E+06	5.60E+06	7.54E+06	5.23E+06	2.98E+06	4.93E+06	2.09E+06	3.63E+06	4.80E+06	3.98E+06	3.39E+07	4.63E+07	3.90E+07
L-cystine	4.65E+03	8.39E+03	8.97E+03	7.00E+03	1.32E+03	9.84E+03	1.51E+03	8.20E+02	1.02E+03	1.92E+03	9.43E+02	1.94E+03	3.06E+03	1.43E+03	5.00E+03	6.30E+04	1.80E+04	4.93E+04
Adenine	7.33E+04	6.98E+04	8.65E+04	8.54E+04	5.68E+04	5.99E+04	4.43E+04	7.70E+04	6.80E+04	9.88E+04	1.18E+05	4.86E+04	9.60E+04	8.80E+04	1.90E+05	1.53E+06	1.22E+06	5.24E+05
Guanine	2.68E+03	5.35E+03	6.27E+03	5.26E+03	4.79E+03	8.40E+03	5.80E+03	3.79E+03	8.84E+03	9.53E+03	7.92E+03	1.52E+04	9.16E+03	1.34E+04	5.37E+04	3.71E+03	5.14E+03	5.09E+03
Cytosine	8.67E+04	4.90E+04	4.99E+04	2.02E+04	6.27E+04	3.50E+04	6.11E+04	6.94E+04	3.31E+04	1.26E+05	1.63E+05	7.73E+04	1.27E+05	1.24E+05	1.09E+05	2.07E+04	1.25E+04	3.65E+04
Thymine	4.52E+04	3.67E+04	2.92E+04	1.71E+04	4.91E+04	4.76E+04	8.41E+04	1.26E+05	9.62E+04	5.41E+04	9.13E+04	6.97E+04	6.07E+04	5.55E+04	5.95E+04	2.04E+04	2.33E+04	3.24E+04
Hypoxanthine	2.59E+05	1.32E+05	1.63E+05	1.24E+05	1.01E+05	8.02E+04	1.60E+05	1.35E+05	1.20E+05	2.28E+05	1.57E+05	6.71E+04	1.84E+05	8.96E+04	2.95E+05	1.74E+05	1.48E+05	2.10E+05
Xanthine	2.60E+04	3.03E+04	3.03E+04	2.42E+04	3.19E+04	2.94E+04	4.80E+04	3.76E+04	4.53E+04	2.43E+04	3.82E+04	2.80E+04	2.98E+04	3.65E+04	2.58E+04	2.94E+04	3.38E+04	3.36E+04
5-Hydroxyisourate	3.41E+06	1.63E+06	1.02E+06	1.10E+06	2.00E+06	1.28E+06	1.29E+06	1.63E+06	6.26E+05	6.55E+06	6.39E+06	9.54E+05	2.66E+06	1.12E+06	1.64E+06	5.29E+04	5.76E+04	1.92E+04
Allantoate	1.45E+05	1.16E+05	1.02E+05	8.42E+04	1.94E+05	1.86E+05	1.28E+05	1.31E+05	1.62E+05	2.32E+05	2.82E+05	1.89E+05	1.59E+05	1.74E+05	1.39E+05	3.57E+05	3.80E+05	2.45E+05
3',5'-Cyclic AMP	2.64E+05	2.92E+04	4.42E+04	1.85E+04	3.12E+05	2.72E+04	2.75E+05	1.93E+05	4.43E+04	3.31E+05	2.20E+05	1.77E+04	1.38E+05	4.06E+04	3.81E+04	0.00E+00	0.00E+00	1.34E+04
5-6-Dihydrothymine	8.38E+06	5.69E+06	5.65E+06	5.59E+06	9.35E+06	6.13E+06	1.03E+07	1.30E+07	1.34E+07	4.15E+06	4.80E+06	3.23E+06	5.00E+06	4.27E+06	6.94E+06	5.52E+05	1.35E+06	3.56E+05
Pyridoxal	4.72E+04	6.71E+04	5.99E+04	8.50E+04	9.72E+04	1.19E+05	3.06E+04	5.23E+04	7.96E+04	4.02E+04	4.08E+04	3.63E+04	5.23E+04	3.94E+04	4.68E+04	4.60E+04	1.93E+04	1.57E+04
Pyridoxamine 5'-phosphate	1.29E+04	2.36E+04	1.14E+04	2.49E+04	1.65E+04	2.67E+04	1.36E+04	1.13E+04	9.52E+03	3.03E+04	2.93E+04	1.49E+04	1.13E+04	8.09E+03	1.19E+04	1.08E+04	1.61E+04	1.06E+04
Nicotinamide	7.58E+05	5.71E+05	4.54E+05	3.12E+05	4.75E+05	4.41E+05	5.04E+05	5.07E+05	4.42E+05	6.95E+05	6.36E+05	2.89E+05	5.63E+05	4.73E+05	6.16E+05	2.53E+06	3.24E+06	1.49E+06
NAD+	6.60E+05	2.59E+05	3.21E+05	1.99E+05	3.98E+05	1.09E+05	2.62E+05	2.70E+05	1.93E+05	6.06E+05	6.52E+05	1.74E+05	5.51E+05	2.01E+05	5.83E+05	6.19E+04	3.83E+04	1.35E+05
Mannitol/Sorbitol/Glucitol/Iditol	2.01E+04	3.20E+04	1.28E+04	2.38E+04	2.10E+04	5.28E+04	2.14E+04	2.97E+04	1.46E+04	7.44E+04	2.42E+04	2.34E+04	1.90E+04	1.47E+04	1.05E+04	1.78E+05	1.67E+05	2.45E+05
D-Rhamnose	4.43E+03	1.35E+04	5.20E+03	5.64E+03	6.78E+03	1.99E+04	1.31E+04	1.96E+04	2.72E+04	8.05E+03	9.40E+03	2.39E+04	2.66E+03	8.76E+03	1.01E+04	1.50E+05	1.46E+05	2.60E+05
Cys-Gly	7.11E+05	5.45E+05	6.28E+05	5.84E+05	8.44E+05	4.73E+05	8.00E+05	9.48E+05	9.04E+05	1.57E+06	1.08E+06	7.87E+05	1.50E+06	8.90E+05	1.53E+06	7.16E+05	5.35E+05	6.95E+05
gamma-L-Glutamyl-L-cysteine	3.74E+04	3.00E+04	2.32E+04	2.76E+04	3.06E+04	1.61E+04	2.95E+04	3.78E+04	2.32E+04	4.10E+04	4.35E+04	1.12E+04	4.73E+04	2.92E+04	4.99E+04	3.72E+04	3.22E+04	8.04E+04
gamma-Glutamyl-Se-methylselenocysteine	3.82E+04	3.43E+04	2.00E+04	3.31E+04	4.57E+04	5.26E+04	3.15E+04	3.60E+04	5.98E+04	4.94E+04	6.37E+04	6.92E+04	4.65E+04	6.67E+04	4.71E+04	1.47E+05	1.27E+05	9.07E+04
S-Adenosyl-L-homocysteine	1.48E+04	8.93E+03	7.18E+03	6.04E+03	6.73E+03	3.06E+03	9.66E+03	9.57E+03	3.51E+03	1.71E+03	1.23E+04	0.00E+00	0.00E+00	1.57E+04	1.07E+04	3.07E+05	3.98E+05	3.29E+05
S-Adenosyl-L-methionine	1.17E+04	1.97E+04	1.39E+04	2.30E+04	1.62E+04	2.32E+04	2.73E+04	3.05E+04	1.48E+03	3.63E+04	3.81E+04	0.00E+00	1.99E+04	2.85E+04	4.43E+04	6.44E+04	2.13E+04	1.01E+05
5-10-Methenyltetrahydrofolate	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	1.23E+05	2.62E+05	4.66E+04
Ornithine	1.72E+05	1.23E+05	1.01E+05	8.24E+04	1.94E+05	1.35E+05	1.51E+05	1.90E+05	1.89E+05	3.89E+05	4.37E+05	1.32E+05	1.71E+05	1.59E+05	1.51E+05	2.99E+05	2.41E+05	2.70E+05
L-Citrulline	4.01E+03	2.20E+03	7.40E+03	0.00E+00	2.59E+03	1.47E+04	0.00E+00	3.30E+03	1.56E+04	1.94E+03	0.00E+00	1.29E+04	5.16E+03	8.80E+03	2.56E+04	1.74E+05	1.70E+05	1.82E+05
Putrescine	5.15E+04	6.42E+04	4.96E+04	7.54E+04	3.44E+04	4.80E+04	8.15E+04	4.81E+04	8.29E+04	1.14E+05	1.14E+05	8.27E+04	1.25E+05	1.08E+05	1.13E+05	3.06E+04	3.21E+04	3.14E+04
Spermidine	7.99E+06	9.45E+06	6.42E+06	9.75E+06	4.56E+06	7.58E+06	9.46E+06	6.61E+06	1.42E+07	1.09E+07	9.83E+06	1.28E+07	1.51E+07	1.20E+07	1.54E+07	4.36E+06	3.97E+06	3.64E+06
Spermine	2.33E+07	3.67E+07	2.75E+07	6.56E+07	3.07E+07	5.62E+07	3.12E+07	3.78E+07	3.17E+07	2.68E+07	3.15E+07	3.10E+07	4.05E+07	5.16E+07	2.77E+07	1.83E+06	3.80E+06	1.94E+06
alpha-D-Glucosamine 1-phosphate	2.90E+05	3.32E+05	1.48E+05	2.41E+05	2.88E+05	2.35E+05	1.46E+05	2.18E+05	4.64E+04	3.38E+05	5.48E+05	8.58E+04	3.26E+05	2.14E+05	1.68E+05	9.86E+04	1.12E+05	1.40E+05
Carnosine	1.20E+05	8.53E+04	8.04E+04	8.59E+04	1.65E+05	1.13E+05	2.72E+05	2.22E+05	1.76E+05	3.83E+05	5.84E+05	1.67E+05	1.95E+05	1.35E+05	1.01E+05	9.36E+04	5.83E+04	3.67E+04
Creatine	1.69E+07	1.73E+07	1.22E+07	1.03E+07	1.95E+07	1.63E+07	1.98E+07	2.60E+07	2.60E+07	6.65E+07	9.01E+07	5.80E+07	6.34E+07	6.34E+07	7.52E+07	6.47E+07	4.70E+07	7.02E+07
Creatinine	1.59E+05	1.27E+05	1.04E+05	1.06E+05	2.41E+05	1.67E+05	2.07E+05	2.10E+05	2.18E+05	6.35E+05	6.46E+05	4.92E+05	5.68E+05	5.66E+05	6.18E+05	1.64E+06	2.37E+06	1.35E+06
4-Acetamidobutanoate	5.68E+03	8.81E+03	9.36E+03	1.22E+04	1.51E+04	9.74E+03	7.05E+03	4.56E+03	0.00E+00	7.02E+03	1.55E+04	6.44E+03	0.00E+00	5.74E+03	1.90E+04	4.29E+05	9.31E+04	1.27E+05
N-Acetyl-L-ornithine	2.07E+03	6.23E+03	1.07E+04	3.07E+03	9.67E+03	4.04E+03	4.90E+03	0.00E+00	6.72E+03	6.96E+03	6.46E+03	1.86E+03	1.41E+03	2.60E+03	1.30E+04	7.75E+04	6.74E+04	3.45E+04
Guanidinoacetate	6.94E+03	1.41E+04	1.02E+04	3.87E+03	8.87E+03	7.95E+03	0.00E+00	1.20E+04	5.08E+03	6.98E+03	9.22E+03	6.90E+03	0.00E+00	0.00E+00	4.11E+03	5.01E+04	3.71E+04	1.45E+04
Indole-3-acetate	3.03E+05	2.23E+05	2.97E+05	2.15E+05	3.99E+05	2.99E+05	3.51E+05	3.26E+05	2.79E+05	3.67E+05	4.49E+05	1.77E+05	3.42E+05	3.07E+05	2.59E+05	1.01E+03	1.59E+03	1.20E+03
Indolepyruvate	2.16E+04	6.91E+04	2.72E+04	4.20E+04	3.40E+04	9.24E+04	2.56E+04	0.00E+00	1.27E+03	8.21E+04	3.37E+04	1.81E+04	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00	0.00E+00
6-Hydroxykynurenic acid	1.24E+04	1.12E+04	2.26E+04	1.26E+04	9.89E+03	4.10E+04	1.56E+04	1.59E+04	1.55E+04	2.03E+04	2.20E+04	7.41E+03	1.09E+04	1.60E+04	2.38E+05	1.01E+04	1.14E+04	1.08E+04
L-Tryptophanamide	2.42E+04	3.06E+04	1.64E+04	1.74E+04	7.86E+03	1.59E+04	1.66E+04	1.18E+04	3.11E+04	3.00E+04	2.54E+04	2.98E+04	2.62E+04	1.64E+04	4.26E+04	5.61E+03	3.03E+03	5.85E+03
Picolinic acid	5.35E+04	6.29E+04	8.45E+04	6.13E+04	9.38E+04	6.84E+04	1.41E+05	1.01E+05	1.17E+05	8.07E+04	1.15E+05	1.04E+05	7.75E+04	1.39E+05	6.95E+04	8.15E+04	1.26E+05	9.66E+04
g-Oxalo-crotonate	1.18E+03	1.62E+03	1.31E+03	1.68E+03	6.45E+02	1.06E+03	0.00E+00	0.00E+00	6.46E+03	0.00E+00	0.00E+00	2.08E+03	2.23E+03	1.00E+03	1.05E+04	3.96E+04	4.77E+04	4.99E+04
Dopamine	2.30E+03	3.41E+03	4.44E+03	2.02E+03	2.39E+03	3.56E+03	2.39E+03	4.03E+03	6.13E+03	3.00E+03	7.73E+03	1.96E+03	5.07E+03	2.05E+03	8.41E+03	2.80E+03	0.00E+00	0.00E+00
Choline	1.30E+07	1.09E+07	4.20E+07	2.63E+07	7.56E+06	6.13E+06	3.24E+07	9.42E+06	1.44E+07	6.30E+07	3.67E+07	5.71E+07	8.03E+07	2.41E+07	3.64E+07	8.90E+06	6.21E+06	9.79E+06
Acetylcholine	1.38E+07	8.08E+06	6.07E+06	3.68E+06	1.37E+07	5.57E+06	8.44E+06	1.30E+07	1.17E+07	1.20E+07	2.02E+07	1.42E+07	1.83E+07	1.78E+07	1.80E+07	2.22E+07	1.38E+07	1.77E+07
L-Carnitine	3.46E+06	5.85E+06	1.37E+06	4.37E+06	2.05E+06	5.13E+06	4.25E+06	6.78E+06	7.07E+06	8.96E+06	1.37E+07	1.68E+07	1.30E+07	1.78E+07	2.09E+07	1.67E+07	2.45E+07	2.25E+07
AC(2:0)	6.69E+06	1.83E+07	4.01E+06	1.26E+07	4.60E+06	1.48E+07	9.84E+06	1.22E+07	7.39E+06	5.95E+07	4.65E+07	3.15E+07	5.16E+07	4.60E+07	2.85E+07	2.33E+07	2.30E+07	2.04E+07
AC(3:0)	2.99E+05	4.41E+05	1.62E+05	3.89E+05	1.27E+05	2.84E+05	3.16E+05	5.32E+05	5.36E+05	1.87E+06	2.50E+06	2.47E+06	1.84E+06	2.77E+06	1.47E+06	1.37E+06	1.16E+06	2.11E+06
AC(4:0)	6.16E+04	1.59E+05	3.05E+04	8.98E+04	3.78E+04	7.02E+04	9.27E+04	6.43E+04	1.78E+05	2.88E+05	3.04E+05	1.68E+06	2.14E+05	1.77E+05	1.27E+06	1.12E+05	1.40E+05	1.02E+05
AC(4-OH)	2.43E+04	3.60E+04	8.51E+03	3.48E+04	1.55E+04	3.12E+04	1.74E+04	1.43E+04	9.72E+03	2.41E+05	1.46E+05	1.53E+05	1.76E+05	2.20E+05	1.77E+05	2.32E+05	3.43E+05	1.85E+05
AC(4-DC)	1.60E+04	2.79E+04	3.55E+03	3.74E+04	9.44E+03	1.62E+04	6.81E+03	1.68E+04	8.10E+03	7.81E+04	5.00E+04	4.69E+04	3.57E+04	5.82E+04	5.39E+04	2.41E+06	2.66E+06	2.38E+06
AC(5:0)	7.38E+04	1.34E+05	4.40E+04	9.33E+04	6.19E+04	1.18E+05	2.02E+05	1.54E+05	1.09E+05	4.91E+05	6.20E+05	4.08E+05	2.94E+05	2.30E+05	2.39E+05	1.35E+05	2.01E+05	1.65E+05
AC(5-OH)	3.52E+04	3.96E+04	1.25E+04	3.18E+04	1.81E+04	4.63E+04	4.69E+04	4.99E+04	2.81E+04	8.72E+04	8.35E+04	6.14E+04	6.02E+04	7.56E+04	4.59E+04	4.59E+05	5.56E+05	1.99E+06
AC(8:0)	3.91E+04	5.74E+04	1.51E+04	5.72E+04	3.89E+04	9.48E+04	1.21E+05	1.27E+05	1.41E+05	2.85E+05	4.67E+05	6.36E+05	2.11E+05	1.98E+05	1.86E+05	4.05E+04	3.26E+04	2.63E+04
AC(10:0)	8.35E+03	1.25E+04	1.52E+03	7.84E+03	8.24E+03	1.48E+04	2.28E+04	2.00E+04	1.55E+04	5.00E+04	1.03E+05	1.12E+05	4.76E+04	4.61E+04	3.21E+04	9.55E+03	1.29E+04	7.88E+03
AC(12:0)	9.78E+03	1.81E+04	2.55E+03	1.28E+04	8.01E+03	1.71E+04	2.10E+04	2.80E+04	2.13E+04	1.03E+05	1.87E+05	2.09E+05	8.23E+04	6.78E+04	6.36E+04	4.49E+03	6.99E+03	6.23E+03
AC(14:0)	3.16E+05	3.58E+05	1.19E+05	2.06E+05	1.93E+05	2.22E+05	1.52E+05	1.45E+05	1.20E+05	5.06E+05	5.15E+05	6.71E+05	3.93E+05	2.95E+05	3.23E+05	1.25E+05	1.19E+05	1.48E+05
AC(14:1)	5.60E+03	9.44E+03	2.95E+03	1.10E+04	4.19E+03	1.18E+04	2.21E+04	1.48E+04	1.31E+04	7.71E+04	1.01E+05	1.31E+05	5.09E+04	4.24E+04	5.62E+04	2.60E+04	3.47E+04	2.56E+04
AC(16:0)	2.94E+06	3.04E+06	7.04E+05	1.75E+06	1.45E+06	2.40E+06	1.92E+06	1.94E+06	1.25E+06	5.18E+06	4.61E+06	4.91E+06	3.61E+06	2.75E+06	3.13E+06	5.82E+06	3.18E+06	3.88E+06
AC(16:1)	1.24E+05	1.89E+05	4.29E+04	9.79E+04	8.02E+04	1.31E+05	1.90E+05	1.91E+05	1.18E+05	3.19E+05	4.01E+05	3.75E+05	2.93E+05	2.95E+05	2.59E+05	1.56E+05	1.28E+05	1.63E+05
AC(18:0)	6.12E+05	7.85E+05	1.26E+05	5.96E+05	6.35E+05	4.89E+05	1.27E+06	7.17E+05	7.13E+05	1.97E+06	1.65E+06	1.96E+06	2.97E+06	3.09E+06	1.98E+06	2.33E+06	1.57E+06	2.19E+06
AC(18:1)	9.49E+05	1.48E+06	1.81E+05	8.52E+05	5.66E+05	1.22E+06	1.52E+06	9.45E+05	6.18E+05	1.67E+06	1.35E+06	1.56E+06	1.57E+06	1.21E+06	1.30E+06	7.82E+06	5.04E+06	5.14E+06
AC(18:2)	6.64E+05	9.91E+05	1.40E+05	5.44E+05	3.68E+05	6.66E+05	1.06E+06	7.35E+05	5.08E+05	1.39E+06	1.25E+06	1.52E+06	1.68E+06	1.40E+06	1.23E+06	3.16E+06	1.93E+06	1.76E+06
AC(20:4)	2.83E+04	5.98E+04	7.77E+03	3.99E+04	1.56E+04	5.48E+04	5.09E+04	3.70E+04	1.61E+04	1.69E+05	1.78E+05	1.47E+05	1.56E+05	1.21E+05	1.28E+05	6.50E+05	4.74E+05	7.69E+05
Sphingosine	4.26E+05	3.31E+05	2.49E+05	2.51E+05	3.59E+05	2.77E+05	3.33E+05	3.87E+05	3.63E+05	4.36E+05	9.21E+05	7.24E+05	8.77E+05	6.94E+05	9.14E+05	2.56E+04	3.39E+04	4.10E+04
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	CmpdID	Pathway	m/z	RT	Polarity
L-alanine	C00041	Amino acids	90.056	0.621	[M+H]+
L-arginine	C00062	Amino acids	175.119	0.614	[M+H]+
L-asparagine	C00152	Amino acids	133.061	0.634	[M+H]+
L-cysteine	C00097	Amino acids	122.027	1.711	[M+H]+
glycine	C00037	Amino acids	76.040	0.652	[M+H]+
L-histidine	C00135	Amino acids	156.077	0.605	[M+H]+
L-leucine/isoluecine	C00123	Amino acids	132.102	0.821	[M+H]+
L-lysine	C00047	Amino acids	147.113	0.598	[M+H]+
L-methionine	C00073	Amino acids	150.058	0.664	[M+H]+
L-phenylalanine	C00079	Amino acids	166.086	1.062	[M+H]+
L-proline	C00148	Amino acids	116.071	0.657	[M+H]+
L-serine	C00065	Amino acids	106.050	0.628	[M+H]+
L-threonine	C00188	Amino acids	120.066	0.627	[M+H]+
L-tryptophan	C00078	Amino acids	205.097	1.695	[M+H]+
L-tyrosine	C00082	Amino acids	182.081	0.665	[M+H]+
L-valine	C00183	Amino acids	118.086	0.693	[M+H]+
L-cystine	C00491	Amino acids	241.029	0.696	[M+H]+
Adenine	C00147	Nucleotides	136.062	0.737	[M+H]+
Guanine	C00242	Nucleotides	152.057	0.707	[M+H]+
Cytosine	C00380	Nucleotides	112.051	0.681	[M+H]+
Thymine	C00178	Nucleotides	127.050	0.659	[M+H]+
Hypoxanthine	C00262	Nucleotides	137.046	0.688	[M+H]+
Xanthine	C00385	Nucleotides	153.039	0.563	[M+H]+
5-Hydroxyisourate	C11821	Nucleotides	185.031	0.625	[M+H]+
Allantoate	C00499	Nucleotides	177.061	0.586	[M+H]+
3',5'-Cyclic AMP	C00575	Nucleotides	330.058	0.761	[M+H]+
5-6-Dihydrothymine	C00906	Nucleotides	129.066	0.655	[M+H]+
Pyridoxal	C00250	Nucleotides	168.065	0.674	[M+H]+
Pyridoxamine 5'-phosphate	C00647	Nucleotides	249.061	1.711	[M+H]+
Nicotinamide	C00153	Nucleotides	123.056	0.754	[M+H]+
NAD+	C00003	Nucleotides	664.114	0.655	[M+H]+
Mannitol/Sorbitol/Glucitol/Iditol	C00392	Other sugars	183.085	0.673	[M+H]+
D-Rhamnose	C01684	Other sugars	165.077	0.634	[M+H]+
Cys-Gly	C01419	GSH homeostasis	179.048	0.692	[M+H]+
gamma-L-Glutamyl-L-cysteine	C00669	Gamma-glutamyls	251.069	0.705	[M+H]+
gamma-Glutamyl-Se-methylselenocysteine	C05695	Gamma-glutamyls	313.031	0.624	[M+H]+
S-Adenosyl-L-homocysteine	C00021	Serine biosynthesis and one-carbon metabolism	385.129	0.653	[M+H]+
S-Adenosyl-L-methionine	C00019	Serine biosynthesis and one-carbon metabolism	399.144	0.617	[M+H]+
5-10-Methenyltetrahydrofolate	C00445	Folate pool (One carbon metabolism)	457.169	0.651	[M+H]+
Ornithine	C01602	Urea cycle	133.097	0.626	[M+H]+
L-Citrulline	C00327	Urea cycle	176.103	0.662	[M+H]+
Putrescine	C00134	Polyamines	89.108	0.559	[M+H]+
Spermidine	C00315	Polyamines	146.165	0.57	[M+H]+
Spermine	C00750	Polyamines	203.223	0.575	[M+H]+
alpha-D-Glucosamine 1-phosphate	C06156	Aminosugars	260.053	0.648	[M+H]+
Carnosine	C00386	Arginine and proline metabolism	227.114	0.593	[M+H]+
Creatine	C00300	Arginine and proline metabolism	132.077	0.639	[M+H]+
Creatinine	C00791	Arginine and proline metabolism	114.066	0.644	[M+H]+
4-Acetamidobutanoate	C02946	Arginine and proline metabolism	146.081	0.651	[M+H]+
N-Acetyl-L-ornithine	C00437	Arginine and proline metabolism	175.108	0.64	[M+H]+
Guanidinoacetate	C00581	Arginine and proline metabolism	118.061	0.669	[M+H]+
Indole-3-acetate	C00954	Indole and Tryptophan	176.071	1.884	[M+H]+
Indolepyruvate	C00331	Indole and Tryptophan	204.063	0.656	[M+H]+
6-Hydroxykynurenic acid	C08480	Indole and Tryptophan	206.045	1.712	[M+H]+
L-Tryptophanamide	C00977	Indole and Tryptophan	204.111	0.549	[M+H]+
Picolinic acid	C10164	Indole and Tryptophan	124.040	0.543	[M+H]+
g-Oxalo-crotonate	C03453	Indole and Tryptophan	159.028	0.677	[M+H]+
Dopamine	C03758	Signaling	154.085	0.666	[M+H]+
Choline	C00114	Glycerophospholipid biosynthesis	104.107	0.623	[M+H]+
Acetylcholine	C01996	Glycerophospholipid biosynthesis	146.117	0.652	[M+H]+
L-Carnitine	C00318	Carnitine and fatty acid metabolsim	162.112	0.627	[M+H]+
AC(2:0)	C02571	Carnitine and fatty acid metabolsim	204.123	0.661	[M+H]+
AC(3:0)	C03017	Carnitine and fatty acid metabolsim	218.139	0.669	[M+H]+
AC(4:0)	C02862	Carnitine and fatty acid metabolsim	232.154	1.685	[M+H]+
AC(4-OH)	HMDB13127	Carnitine and fatty acid metabolsim	248.149	0.662	[M+H]+
AC(4-DC)	HMDB13133	Carnitine and fatty acid metabolsim	262.129	0.666	[M+H]+
AC(5:0)	C20826	Carnitine and fatty acid metabolsim	246.170	1.715	[M+H]+
AC(5-OH)	ac107	Carnitine and fatty acid metabolsim	262.165	0.669	[M+H]+
AC(8:0)	HMDB00791	Carnitine and fatty acid metabolsim	288.217	1.838	[M+H]+
AC(10:0)	HMDB00651	Carnitine and fatty acid metabolsim	316.248	1.889	[M+H]+
AC(12:0)	HMDB02250	Carnitine and fatty acid metabolsim	344.279	1.932	[M+H]+
AC(14:0)	HMDB05066	Carnitine and fatty acid metabolsim	372.310	1.991	[M+H]+
AC(14:1)	HMDB13329	Carnitine and fatty acid metabolsim	370.295	1.96	[M+H]+
AC(16:0)	C02990	Carnitine and fatty acid metabolsim	400.342	2.044	[M+H]+
AC(16:1)	HMDB06317	Carnitine and fatty acid metabolsim	398.327	1.994	[M+H]+
AC(18:0)	HMDB00848	Carnitine and fatty acid metabolsim	428.373	2.112	[M+H]+
AC(18:1)	HMDB06351	Carnitine and fatty acid metabolsim	426.357	2.052	[M+H]+
AC(18:2)	HMDB01064	Carnitine and fatty acid metabolsim	424.342	2.017	[M+H]+
AC(20:4)	ac125	Carnitine and fatty acid metabolsim	448.341	2.005	[M+H]+
Sphingosine	C00319	Sphingolipid biosynthesis	300.289	1.979	[M+H]+
METABOLITES_END
#END