#METABOLOMICS WORKBENCH manikana_20251010_075853 DATATRACK_ID:6548 STUDY_ID:ST004277 ANALYSIS_ID:AN007116 PROJECT_ID:PR002703
VERSION             	1
CREATED_ON             	October 13, 2025, 2:22 pm
#PROJECT
PR:PROJECT_TITLE                 	Limitations in PPAR⍺-dependent mitochondrial programming restrain the
PR:PROJECT_TITLE                 	differentiation of human stem cell-derived β cells
PR:PROJECT_SUMMARY               	Pluripotent stem cell (SC)-derived islets offer hope as a renewable source for
PR:PROJECT_SUMMARY               	β cell replacement for type 1 diabetes (T1D), yet functional and metabolic
PR:PROJECT_SUMMARY               	immaturity may limit their long-term therapeutic potential. Here, we show that
PR:PROJECT_SUMMARY               	limitations in mitochondrial transcriptional programming impede the formation of
PR:PROJECT_SUMMARY               	SC-derived β (SC-β) cells. Utilizing transcriptomic profiling, assessments of
PR:PROJECT_SUMMARY               	chromatin accessibility, mitochondrial phenotyping, and lipidomics analyses, we
PR:PROJECT_SUMMARY               	observed that SC-β cells exhibit reduced oxidative and mitochondrial fatty acid
PR:PROJECT_SUMMARY               	metabolism compared to primary human islets that are related to limitations in
PR:PROJECT_SUMMARY               	key mitochondrial transcriptional networks. Surprisingly, we find that
PR:PROJECT_SUMMARY               	reductions in glucose-stimulated mitochondrial respiration in SC-islets were not
PR:PROJECT_SUMMARY               	associated with alterations in mitochondrial mass, structure, or genome
PR:PROJECT_SUMMARY               	integrity. In contrast, SC-islets show limited expression of targets of PPAR⍺,
PR:PROJECT_SUMMARY               	which regulate mitochondrial programming, yet whose functions in β cell
PR:PROJECT_SUMMARY               	differentiation are unknown. Importantly, treatment with WY14643, a potent
PR:PROJECT_SUMMARY               	PPAR⍺ agonist, induced expression of mitochondrial targets, improved insulin
PR:PROJECT_SUMMARY               	secretion, and increased the formation of SC-β cells both in vitro and
PR:PROJECT_SUMMARY               	following transplantation. Thus, PPAR⍺-dependent mitochondrial programming
PR:PROJECT_SUMMARY               	promotes the differentiation of SC-β cells and may be a promising target to
PR:PROJECT_SUMMARY               	improve β cell replacement efforts for T1D.
PR:INSTITUTE                     	University of Michigan
PR:DEPARTMENT                    	Division of Metabolism, Endocrinology and Diabetes and Department of Internal
PR:DEPARTMENT                    	Medicine
PR:LABORATORY                    	Scott A. Soleimanpour
PR:LAST_NAME                     	Arnipalli
PR:FIRST_NAME                    	Manikanta
PR:ADDRESS                       	3815-301B Green Brier apt, Ann Arbor, michigan, 48105, USA
PR:EMAIL                         	manikana@umich.edu
PR:PHONE                         	734-272-8779
#STUDY
ST:STUDY_TITLE                   	Limitations in PPAR⍺-dependent mitochondrial programming restrain the
ST:STUDY_TITLE                   	differentiation of human stem cell-derived β cells
ST:STUDY_SUMMARY                 	Pluripotent stem cell (SC)-derived islets offer hope as a renewable source for
ST:STUDY_SUMMARY                 	β cell replacement for type 1 diabetes (T1D), yet functional and metabolic
ST:STUDY_SUMMARY                 	immaturity may limit their long-term therapeutic potential. Here, we show that
ST:STUDY_SUMMARY                 	limitations in mitochondrial transcriptional programming impede the formation of
ST:STUDY_SUMMARY                 	SC-derived β (SC-β) cells. Utilizing transcriptomic profiling, assessments of
ST:STUDY_SUMMARY                 	chromatin accessibility, mitochondrial phenotyping, and lipidomics analyses, we
ST:STUDY_SUMMARY                 	observed that SC-β cells exhibit reduced oxidative and mitochondrial fatty acid
ST:STUDY_SUMMARY                 	metabolism compared to primary human islets that are related to limitations in
ST:STUDY_SUMMARY                 	key mitochondrial transcriptional networks. Surprisingly, we find that
ST:STUDY_SUMMARY                 	reductions in glucose-stimulated mitochondrial respiration in SC-islets were not
ST:STUDY_SUMMARY                 	associated with alterations in mitochondrial mass, structure, or genome
ST:STUDY_SUMMARY                 	integrity. In contrast, SC-islets show limited expression of targets of PPAR⍺,
ST:STUDY_SUMMARY                 	which regulate mitochondrial programming, yet whose functions in β cell
ST:STUDY_SUMMARY                 	differentiation are unknown. Importantly, treatment with WY14643, a potent
ST:STUDY_SUMMARY                 	PPAR⍺ agonist, induced expression of mitochondrial targets, improved insulin
ST:STUDY_SUMMARY                 	secretion, and increased the formation of SC-β cells both in vitro and
ST:STUDY_SUMMARY                 	following transplantation. Thus, PPAR⍺-dependent mitochondrial programming
ST:STUDY_SUMMARY                 	promotes the differentiation of SC-β cells and may be a promising target to
ST:STUDY_SUMMARY                 	improve β cell replacement efforts for T1D.
ST:INSTITUTE                     	University of Michigan
ST:DEPARTMENT                    	Division of Metabolism, Endocrinology and Diabetes and Department of Internal
ST:DEPARTMENT                    	Medicine
ST:LABORATORY                    	Scott A. Soleimanpour
ST:LAST_NAME                     	Arnipalli
ST:FIRST_NAME                    	Manikanta
ST:ADDRESS                       	3815-301B Green Brier apt, Ann Arbor, michigan, 48105, USA
ST:EMAIL                         	manikana@umich.edu
ST:PHONE                         	734-272-8779
#SUBJECT
SU:SUBJECT_TYPE                  	Human
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           	-	10-04-22 Human islets 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-04-22 Human islets 5.5mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-04-22 Human islets 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-04-22 Human islets 20mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-07-22 Human islets 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-07-22 Human islets 5.5mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-07-22 Human islets 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-07-22 Human islets 20mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-15-22 Human islets 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-15-22 Human islets 5.5mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-15-22 Human islets 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-15-22 Human islets 20mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-28-22 Alberta Human islets 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-28-22 Alberta Human islets 5.5mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-28-22 Alberta Human islets 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-28-22 Alberta Human islets 20mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-28-22 Prodo Human islets 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-28-22 Prodo Human islets 5.5mM .d
SUBJECT_SAMPLE_FACTORS           	-	10-28-22 Prodo Human islets 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-28-22 Prodo Human islets 20mM .d
SUBJECT_SAMPLE_FACTORS           	-	AS_A_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=AS_A .d
SUBJECT_SAMPLE_FACTORS           	-	AS_B_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=AS_B .d
SUBJECT_SAMPLE_FACTORS           	-	Blank_01_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_01 .d
SUBJECT_SAMPLE_FACTORS           	-	Blank_02_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_02 .d
SUBJECT_SAMPLE_FACTORS           	-	Blank_04_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_04 .d
SUBJECT_SAMPLE_FACTORS           	-	pool_01_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_01 .d
SUBJECT_SAMPLE_FACTORS           	-	pool_02_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_02 .d
SUBJECT_SAMPLE_FACTORS           	-	pool_03_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_03 .d
SUBJECT_SAMPLE_FACTORS           	-	pool_04_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_04 .d
SUBJECT_SAMPLE_FACTORS           	-	pool_05_human	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_05 .d
SUBJECT_SAMPLE_FACTORS           	-	7/22/22 S1 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=7-22-22 S1 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	7/24/22 S2 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=7-24-22 S2 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	7/26/22 S3 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=7-26-22 S3 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	7/30/22 S4 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=7-30-22 S4 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	8/06/22 S5 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=8-06-22 S5 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	8/15/22 S6 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=8-15-22 S6 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/10/22 S1 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-10-22 S1 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/12/22 S2 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-12-22 S2 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/14/22 S1 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-14-22 S1 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/14/22 S3 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-14-22 S3 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/16/22 S2 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-16-22 S2 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/18/22 S3 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-18-22 S3 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/18/22 S4 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-18-22 S4 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/22/22 S4 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-22-22 S4 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/25/22 S5 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-25-22 S5 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	9/29/22 S5 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=9-29-22 S5 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	10/04/22 S6 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-04-22 S6 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	10/08/22 S6 metabolomics 20mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	RAW_FILE_NAME(Raw file name)=10-08-22 S6 metabolomics 20mM.d
SUBJECT_SAMPLE_FACTORS           	-	AS_A_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=AS_A.d
SUBJECT_SAMPLE_FACTORS           	-	AS_B_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=AS_B.d
SUBJECT_SAMPLE_FACTORS           	-	bank_04_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=bank_04.d
SUBJECT_SAMPLE_FACTORS           	-	Blank_01_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_01.d
SUBJECT_SAMPLE_FACTORS           	-	Blank_02_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_02.d
SUBJECT_SAMPLE_FACTORS           	-	Blank_03_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=Blank_03.d
SUBJECT_SAMPLE_FACTORS           	-	pool_01_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_01.d
SUBJECT_SAMPLE_FACTORS           	-	pool_02_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_02.d
SUBJECT_SAMPLE_FACTORS           	-	pool_03_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_03.d
SUBJECT_SAMPLE_FACTORS           	-	pool_04_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_04.d
SUBJECT_SAMPLE_FACTORS           	-	pool_05_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_05.d
SUBJECT_SAMPLE_FACTORS           	-	pool_06_mouse_high	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	RAW_FILE_NAME(Raw file name)=pool_06.d
SUBJECT_SAMPLE_FACTORS           	-	7-22-22 S1 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=7-22-22 S1 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	7-24-22 S2 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=7-24-22 S2 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	7-26-22 S3 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=7-26-22 S3 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	7-30-22 S4 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=7-30-22 S4 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	8-06-22 S5 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=8-06-22 S5 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	8-15-22 S6 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=8-15-22 S6 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-10-22 S1 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-10-22 S1 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-12-22 S2 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-12-22 S2 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-14-22 S1 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-14-22 S1 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-14-22 S3 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-14-22 S3 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-16-22 S2 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-16-22 S2 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-18-22 S3 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-18-22 S3 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-18-22 S4 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-18-22 S4 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-22-22 S4 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-22-22 S4 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-25-22 S5 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-25-22 S5 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	9-29-22 S5 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=9-29-22 S5 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	10-04-22 S6 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-04-22 S6 metabolomics 5.5mM.d
SUBJECT_SAMPLE_FACTORS           	-	10-08-22 S6 metabolomics 5.5mM	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	RAW_FILE_NAME(Raw file name)=10-08-22 S6 metabolomics 5.5mM.d
#COLLECTION
CO:COLLECTION_SUMMARY            	Cells were cultured according to Hogrebe et al4. On the day of sample
CO:COLLECTION_SUMMARY            	collection, samples were incubated in MCDB 131 with 10.5 g BSA, 5.2 mL GlutaMAX,
CO:COLLECTION_SUMMARY            	5.2 mL P/S, 5 mg heparin, 5.2 mL MEM nonessential amino acids (Corning,
CO:COLLECTION_SUMMARY            	20–025-CI), 84 μg ZnSO4 (MilliporeSigma, 10883), 523 μL Trace Elements A
CO:COLLECTION_SUMMARY            	(Corning, 25–021-CI), and 523 μL Trace Elements B (Corning, 25–022-CI) and
CO:COLLECTION_SUMMARY            	either 5.5mM or 20mM glucose for 3 hr. Media was removed, and the cells were
CO:COLLECTION_SUMMARY            	washed 1X with ice-cold 150 mM ammonium acetate in LC-MS grade water. Cells were
CO:COLLECTION_SUMMARY            	harvested with 200 μL of ice-cold methanol and frozen at -80°C until sample
CO:COLLECTION_SUMMARY            	preparation.
CO:SAMPLE_TYPE                   	Pancreas
#TREATMENT
TR:TREATMENT_SUMMARY             	Treatment not applied.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	For extraction, 200 μL of cold water was added to the cells and the cells were
SP:SAMPLEPREP_SUMMARY            	scraped. The resulting homogenate was sonicated on ice for 10 sec. 400 μL of
SP:SAMPLEPREP_SUMMARY            	chloroform was added. Samples were centrifuged at 17,000 x g for 10 min and the
SP:SAMPLEPREP_SUMMARY            	resulting top layer was collected and taken to dryness under nitrogen. Samples
SP:SAMPLEPREP_SUMMARY            	were reconstituted in 30 μL of 2:1 ACN: water, filtered and 5 μL were injected
SP:SAMPLEPREP_SUMMARY            	for analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Metabolites were separated on an InfinityLab Poroshell120 HILIC-Z, 2.7 μm, 2.1
CH:CHROMATOGRAPHY_SUMMARY        	× 150 mm column (Agilent Technologies, CA, USA). Mobile phase A was composed of
CH:CHROMATOGRAPHY_SUMMARY        	90% 10 mM ammonium acetate pH 9.0 with ammonia, 10% acetonitrile (ACN) with 2.5
CH:CHROMATOGRAPHY_SUMMARY        	uM InfinityLab Deactivator Additive (Agilent Technologies, CA, USA) and mobile
CH:CHROMATOGRAPHY_SUMMARY        	phase B was composed of 15% 10 mM ammonium acetate pH 9.0 with ammonia, 85%
CH:CHROMATOGRAPHY_SUMMARY        	acetonitrile with 2.5 uM InfinityLab Deactivator Additive. The flow rate was
CH:CHROMATOGRAPHY_SUMMARY        	0.25 mL/min, and the gradient was as follows: 0-2 min at 95 % B, 2-5 min at 95%
CH:CHROMATOGRAPHY_SUMMARY        	B, 5-5.5 min at 86%, 5.5-8.5 min at 86% B, 8.5-9 min at 84% B, 9-14 min at 84%
CH:CHROMATOGRAPHY_SUMMARY        	B, 14-17 min at 80% B, 17-23 min at 60% B, 23-26 min at 60% B, 26-27 min at 95%
CH:CHROMATOGRAPHY_SUMMARY        	B, and 27-35 min at 95% B. Column compartment temperature was kept at 25 °C.
CH:CHROMATOGRAPHY_SUMMARY        	Data were acquired in negative mode.
CH:CHROMATOGRAPHY_TYPE           	HILIC
CH:INSTRUMENT_NAME               	Agilent 1290 Infinity II
CH:COLUMN_NAME                   	Agilent Poroshell120 HILIC-Z (150 x 2.1 mm, 2.7 μm)
CH:SOLVENT_A                     	90% Water/10% Acetonitrile; 10 mM ammonium acetate pH 9.0 with ammonia;
CH:SOLVENT_A                     	2.5 μM InfinityLab Deactivator Additive
CH:SOLVENT_B                     	85% Acetonitrile/15% Water; 10 mM Ammonium acetate pH 9.0 with ammonia; 2.5 μM
CH:SOLVENT_B                     	InfinityLab Deactivator Additive
CH:FLOW_GRADIENT                 	0-2 min at 95 % B, 2-5 min at 95% B, 5-5.5 min at 86%, 5.5-8.5 min at 86% B,
CH:FLOW_GRADIENT                 	8.5-9 min at 84% B, 9-14 min at 84% B, 14-17 min at 80% B, 17-23 min at 60% B,
CH:FLOW_GRADIENT                 	23-26 min at 60% B, 26-27 min at 95% B, and 27-35 min at 95% B
CH:FLOW_RATE                     	0.25 mL/min
CH:COLUMN_TEMPERATURE            	25°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Agilent 6546 QTOF
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The gas temperature was set at 225°C, the drying gas flow at 10 L/min, the
MS:MS_COMMENTS                   	nebulizer at 40 psi, the sheath gas temperature at 300°C, and the sheath gas
MS:MS_COMMENTS                   	flow at 12 L/min. The Fragmentor (Agilent) was set at 125 V, the skimmer at 65
MS:MS_COMMENTS                   	V, and VCap at 3,000 V.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	peak area
MS_METABOLITE_DATA_START
Samples	10-04-22 Human islets 5.5mM	10-04-22 Human islets 20mM	10-07-22 Human islets 5.5mM	10-07-22 Human islets 20mM	10-15-22 Human islets 5.5mM	10-15-22 Human islets 20mM	10-28-22 Alberta Human islets 5.5mM	10-28-22 Alberta Human islets 20mM	10-28-22 Prodo Human islets 5.5mM	10-28-22 Prodo Human islets 20mM	AS_A_human	AS_B_human	Blank_01_human	Blank_02_human	Blank_04_human	pool_01_human	pool_02_human	pool_03_human	pool_04_human	pool_05_human	7/22/22 S1 metabolomics 20mM	7/24/22 S2 metabolomics 20mM	7/26/22 S3 metabolomics 20mM	7/30/22 S4 metabolomics 20mM	8/06/22 S5 metabolomics 20mM	8/15/22 S6 metabolomics 20mM	9/10/22 S1 metabolomics 20mM	9/12/22 S2 metabolomics 20mM	9/14/22 S1 metabolomics 20mM	9/14/22 S3 metabolomics 20mM	9/16/22 S2 metabolomics 20mM	9/18/22 S3 metabolomics 20mM	9/18/22 S4 metabolomics 20mM	9/22/22 S4 metabolomics 20mM	9/25/22 S5 metabolomics 20mM	9/29/22 S5 metabolomics 20mM	10/04/22 S6 metabolomics 20mM	10/08/22 S6 metabolomics 20mM	AS_A_mouse_high	AS_B_mouse_high	bank_04_mouse_high	Blank_01_mouse_high	Blank_02_mouse_high	Blank_03_mouse_high	pool_01_mouse_high	pool_02_mouse_high	pool_03_mouse_high	pool_04_mouse_high	pool_05_mouse_high	pool_06_mouse_high	7-22-22 S1 metabolomics 5.5mM	7-24-22 S2 metabolomics 5.5mM	7-26-22 S3 metabolomics 5.5mM	7-30-22 S4 metabolomics 5.5mM	8-06-22 S5 metabolomics 5.5mM	8-15-22 S6 metabolomics 5.5mM	9-10-22 S1 metabolomics 5.5mM	9-12-22 S2 metabolomics 5.5mM	9-14-22 S1 metabolomics 5.5mM	9-14-22 S3 metabolomics 5.5mM	9-16-22 S2 metabolomics 5.5mM	9-18-22 S3 metabolomics 5.5mM	9-18-22 S4 metabolomics 5.5mM	9-22-22 S4 metabolomics 5.5mM	9-25-22 S5 metabolomics 5.5mM	9-29-22 S5 metabolomics 5.5mM	10-04-22 S6 metabolomics 5.5mM	10-08-22 S6 metabolomics 5.5mM
Factors	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human | Sample source:Pancreas | Glucose Concentration:High	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human_Control | Sample source:Pancreas | Glucose Concentration:NA	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low	Sample type:Human | Sample source:Pancreas | Glucose Concentration:Low
pyruvate	455784.4795	679676.626	155855.3258	415784.8602	341914.7222	565886.6725	1614991.588	2712169.303	1422524.714	1457275.574	229504.5778	48839.45527				615769.1826	1255280.171	1220800.666	992916.2423	398768.9792	861748.9777	1106577.866	853503.8112	893218.5348	742573.534	788104.23	1150945.686	1155665.847	1166335.407	1328020.909	1101067.001	958352.5112	1133339.082	1009461.439	887031.7048	1025917.897	1241834.409	954150.6447	568294.1146	1969.86325	1641.946907	3431.856712		64658.11758	1004367.775	1187231.963	1441150.509	1255139.534	1272028.496	1242996.418	1176824	1125966	1016052	877673	910850	819878	1612538	1201055	1526654	1443730	1241843	985581	1175706	1312288	1226898	851643	1001114	1197163
pyruvate+3 IS (13C3)	73462.37107	82944.70625	77961.93614	73994.08187	74384.65009	86759.02947	23504.71633	19738.96574	31523.46392	32601.5992						14746.4456	30511.87305	30632.48941	28822.23584	4728.030842	70541.06578	96176.94325	116588.5582	197440.214	52643.78858	98352.05959	49063.16293	90982.16749	90278.74802	139924.7102	56260.20349	116912.9039	132031.814	92192.51639	27371.36946	25109.62124	42003.45212	82151.43212							68248.93004	80900.44562	93953.6342	80674.25841	75769.75416	65266.66996	94110	81433	140874	157348	63688	117148	48180	78677	113638	133383	37901	104341	101219	89291	38936	41178	48191	120906
L-Lactate	1367023.643	1925288.113	466652.8814	779458.6298	254177.0653	690653.6738	1560627.268		347886.1284	170348.0659	233196.4705	2518041.146	3526.634758	43599.20793	80655.23765	1386380.805	1438428.471	1080200.493	1185896.299	218642.1292	7164932.869	7974660.13	8869494.15	9920897.64	7489549.967	4856708.869	7919534.467	4558064.433	3922743.46	8165021.81	4389591.33	8870065.051	10038515.49	9124762.661	5808860.364	9891550.731	5144958.736	9621623.473	435123.2522	3229702.558	24051.85622	37689.38616	37955.41765	84986.40106	8251810.925	5715092.229	7996013.34	9462017.095	6088909.498	4426618.523	7316045	7890982	7887627	9106043	10565875	6348319	8842140	8361339	9458357	9644724	10836322	8369765	8910845	9596567	13414308	7970417	8517764	12021968
L-Lactate+3 IS (13C3)	367865.8035	451008.2633	485412.5222	274400.9966	393145.0121	433036.0581	93180.15389	4986.19029	165878.4788	133975.7646						345932.2353	241580.0569	151711.2295	149351.8556	27946.2125	165507.3501	115640.8244	116273.9416	183417.3906	67145.66639	102442.0938	55366.70482	58166.68173	119897.1219	76820.52085	64044.9666	257485.6274	69681.24136	52582.43822	46548.84262	34790.88175	40266.75336	139176.183							99156.80277	154146.0318	140000.0318	86327.4141	132396.0062	22380.35048	142246	135459	112157	146544	92812	152460	47659	85124	81355	78051	46716	162083	110036	107533	74641	63417	71207	69428
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	RT	KEGG_ID
pyruvate	2.54897526	C00022
pyruvate+3 IS (13C3)	2.54897526
L-Lactate	4.230050995	C00186
L-Lactate+3 IS (13C3)	4.230050995
METABOLITES_END
#END