#METABOLOMICS WORKBENCH Gilles_Perera_Lab_20250320_111111 DATATRACK_ID:5761 STUDY_ID:ST003847 ANALYSIS_ID:AN006322 PROJECT_ID:PR002405
VERSION             	1
CREATED_ON             	April 7, 2025, 7:01 pm
#PROJECT
PR:PROJECT_TITLE                 	Differential acquisition of extracellular lipid correlates with pancreatic
PR:PROJECT_TITLE                 	cancer subtype and metastatic tropism
PR:PROJECT_TYPE                  	Metabolomics
PR:PROJECT_SUMMARY               	Pancreatic ductal adenocarcinoma (PDAC) stands as an aggressive disease, ranking
PR:PROJECT_SUMMARY               	at the third leading cause of cancer-related death with a 5-year overall
PR:PROJECT_SUMMARY               	survival rate of less than 5%. Recent studies have identified two major subtypes
PR:PROJECT_SUMMARY               	of pancreatic cancer, basal and classical, which predict patient prognosis, with
PR:PROJECT_SUMMARY               	basal tumors corresponding to more aggressive disease. While transcriptional
PR:PROJECT_SUMMARY               	signatures define basal and classical tumors and cell lines, little is known
PR:PROJECT_SUMMARY               	regarding the metabolic vulnerabilities linked to the basal versus classical
PR:PROJECT_SUMMARY               	state. Using unbiased computational interrogation to uncover metabolic genes
PR:PROJECT_SUMMARY               	correlating with known markers of classical PDAC, we identified high expression
PR:PROJECT_SUMMARY               	of Proprotein Convertase Subtilisin/Kexin type-9 (PCSK9) – a negative
PR:PROJECT_SUMMARY               	regulator of receptor mediated uptake of cholesterol-containing low-density
PR:PROJECT_SUMMARY               	lipoprotein (LDL) – as correlating with several established markers of
PR:PROJECT_SUMMARY               	classical PDAC. In contrast, PCSK9 expression is suppressed in basal PDAC.
PR:PROJECT_SUMMARY               	Accordingly, basal PDAC cells uptake high levels of LDL, are sensitive to LDL
PR:PROJECT_SUMMARY               	depletion and show higher dependence on cholesterol uptake, relative to
PR:PROJECT_SUMMARY               	classical PDAC. As lipoproteins are produced in the liver, we hypothesized that
PR:PROJECT_SUMMARY               	basal PDAC cells might possess a greater propensity to seed and thrive within
PR:PROJECT_SUMMARY               	liver tissues. Conversely, basal cells demonstrated a robust ability to grow
PR:PROJECT_SUMMARY               	within the liver but faced challenges in invading the lungs whereas classical
PR:PROJECT_SUMMARY               	cells harbour the opposite phenotype. Moreover, we observed that patients with
PR:PROJECT_SUMMARY               	liver metastasis alone had weaker PCSK9 staining at the primary tumor site
PR:PROJECT_SUMMARY               	compared to those with lung metastasis only. This observation suggests a
PR:PROJECT_SUMMARY               	potential influence of cholesterol metabolism in the primary tumor on the
PR:PROJECT_SUMMARY               	tropism of metastatic sites. Modulating PCSK9 levels in pancreatic cancer cells
PR:PROJECT_SUMMARY               	with low expression of this protein resulted in reduced LDL uptake and a shift
PR:PROJECT_SUMMARY               	towards the cholesterol biosynthesis pathway. More importantly, overexpressing
PR:PROJECT_SUMMARY               	PCSK9 not only reduced the liver metastasis burden but also increased lung
PR:PROJECT_SUMMARY               	metastasis in liver-tropic cell lines. Lastly, we found that PCSK9 expression
PR:PROJECT_SUMMARY               	and protein abundance remained low in liver metastatic lesions and high in lung
PR:PROJECT_SUMMARY               	metastasis lesions. This observation suggests that cancer cells maintain
PR:PROJECT_SUMMARY               	distinct cholesterol metabolism profiles in metastatic lesions. Ongoing studies
PR:PROJECT_SUMMARY               	will interrogate the role of cholesterol uptake and utilization in liver
PR:PROJECT_SUMMARY               	metastasis , and uncover unique features and vulnerabilities of the most
PR:PROJECT_SUMMARY               	aggressive variant of PDAC.
PR:INSTITUTE                     	University of California, San Francisco
PR:DEPARTMENT                    	Anatomy
PR:LABORATORY                    	Perera
PR:LAST_NAME                     	Rademaker
PR:FIRST_NAME                    	Gilles
PR:ADDRESS                       	513 Parnassus avenue, San Francisco, CA, 94143, USA
PR:EMAIL                         	gilles.rademaker@ucsf.edu
PR:PHONE                         	4155024290
#STUDY
ST:STUDY_TITLE                   	Differential acquisition of extracellular lipid correlates with pancreatic
ST:STUDY_TITLE                   	cancer subtype and metastatic tropism
ST:STUDY_TYPE                    	Metabolomics
ST:STUDY_SUMMARY                 	Pancreatic ductal adenocarcinoma (PDAC) stands as an aggressive disease, ranking
ST:STUDY_SUMMARY                 	at the third leading cause of cancer-related death with a 5-year overall
ST:STUDY_SUMMARY                 	survival rate of less than 5%. Recent studies have identified two major subtypes
ST:STUDY_SUMMARY                 	of pancreatic cancer, basal and classical, which predict patient prognosis, with
ST:STUDY_SUMMARY                 	basal tumors corresponding to more aggressive disease. While transcriptional
ST:STUDY_SUMMARY                 	signatures define basal and classical tumors and cell lines, little is known
ST:STUDY_SUMMARY                 	regarding the metabolic vulnerabilities linked to the basal versus classical
ST:STUDY_SUMMARY                 	state. Using unbiased computational interrogation to uncover metabolic genes
ST:STUDY_SUMMARY                 	correlating with known markers of classical PDAC, we identified high expression
ST:STUDY_SUMMARY                 	of Proprotein Convertase Subtilisin/Kexin type-9 (PCSK9) – a negative
ST:STUDY_SUMMARY                 	regulator of receptor mediated uptake of cholesterol-containing low-density
ST:STUDY_SUMMARY                 	lipoprotein (LDL) – as correlating with several established markers of
ST:STUDY_SUMMARY                 	classical PDAC. In contrast, PCSK9 expression is suppressed in basal PDAC.
ST:STUDY_SUMMARY                 	Accordingly, basal PDAC cells uptake high levels of LDL, are sensitive to LDL
ST:STUDY_SUMMARY                 	depletion and show higher dependence on cholesterol uptake, relative to
ST:STUDY_SUMMARY                 	classical PDAC. As lipoproteins are produced in the liver, we hypothesized that
ST:STUDY_SUMMARY                 	basal PDAC cells might possess a greater propensity to seed and thrive within
ST:STUDY_SUMMARY                 	liver tissues. Conversely, basal cells demonstrated a robust ability to grow
ST:STUDY_SUMMARY                 	within the liver but faced challenges in invading the lungs whereas classical
ST:STUDY_SUMMARY                 	cells harbour the opposite phenotype. Moreover, we observed that patients with
ST:STUDY_SUMMARY                 	liver metastasis alone had weaker PCSK9 staining at the primary tumor site
ST:STUDY_SUMMARY                 	compared to those with lung metastasis only. This observation suggests a
ST:STUDY_SUMMARY                 	potential influence of cholesterol metabolism in the primary tumor on the
ST:STUDY_SUMMARY                 	tropism of metastatic sites. Modulating PCSK9 levels in pancreatic cancer cells
ST:STUDY_SUMMARY                 	with low expression of this protein resulted in reduced LDL uptake and a shift
ST:STUDY_SUMMARY                 	towards the cholesterol biosynthesis pathway. More importantly, overexpressing
ST:STUDY_SUMMARY                 	PCSK9 not only reduced the liver metastasis burden but also increased lung
ST:STUDY_SUMMARY                 	metastasis in liver-tropic cell lines. Lastly, we found that PCSK9 expression
ST:STUDY_SUMMARY                 	and protein abundance remained low in liver metastatic lesions and high in lung
ST:STUDY_SUMMARY                 	metastasis lesions. This observation suggests that cancer cells maintain
ST:STUDY_SUMMARY                 	distinct cholesterol metabolism profiles in metastatic lesions. Ongoing studies
ST:STUDY_SUMMARY                 	will interrogate the role of cholesterol uptake and utilization in liver
ST:STUDY_SUMMARY                 	metastasis , and uncover unique features and vulnerabilities of the most
ST:STUDY_SUMMARY                 	aggressive variant of PDAC.
ST:INSTITUTE                     	University of California, San Francisco
ST:DEPARTMENT                    	Anatomy
ST:LABORATORY                    	Perera
ST:LAST_NAME                     	Rademaker
ST:FIRST_NAME                    	Gilles
ST:ADDRESS                       	513 Parnassus avenue, San Francisco, CA, 94143, USA
ST:EMAIL                         	gilles.rademaker@ucsf.edu
ST:PHONE                         	4155024290
#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           	-	1	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_01
SUBJECT_SAMPLE_FACTORS           	-	2	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_02
SUBJECT_SAMPLE_FACTORS           	-	3	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_03
SUBJECT_SAMPLE_FACTORS           	-	4	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Replicate=4; RAW_FILE_NAME(Raw File name)=Cell_UCSF_04
SUBJECT_SAMPLE_FACTORS           	-	5	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Replicate=5; RAW_FILE_NAME(Raw File name)=Cell_UCSF_05
SUBJECT_SAMPLE_FACTORS           	-	6	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_06
SUBJECT_SAMPLE_FACTORS           	-	7	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_07
SUBJECT_SAMPLE_FACTORS           	-	8	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_08
SUBJECT_SAMPLE_FACTORS           	-	9	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Replicate=4; RAW_FILE_NAME(Raw File name)=Cell_UCSF_09
SUBJECT_SAMPLE_FACTORS           	-	10	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Replicate=5; RAW_FILE_NAME(Raw File name)=Cell_UCSF_10
SUBJECT_SAMPLE_FACTORS           	-	11	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_11
SUBJECT_SAMPLE_FACTORS           	-	12	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_12
SUBJECT_SAMPLE_FACTORS           	-	13	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_13
SUBJECT_SAMPLE_FACTORS           	-	14	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Replicate=4; RAW_FILE_NAME(Raw File name)=Cell_UCSF_14
SUBJECT_SAMPLE_FACTORS           	-	15	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Replicate=5; RAW_FILE_NAME(Raw File name)=Cell_UCSF_15
SUBJECT_SAMPLE_FACTORS           	-	16	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_16
SUBJECT_SAMPLE_FACTORS           	-	17	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_17
SUBJECT_SAMPLE_FACTORS           	-	18	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_18
SUBJECT_SAMPLE_FACTORS           	-	19	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Replicate=4; RAW_FILE_NAME(Raw File name)=Cell_UCSF_19
SUBJECT_SAMPLE_FACTORS           	-	20	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Replicate=5; RAW_FILE_NAME(Raw File name)=Cell_UCSF_20
SUBJECT_SAMPLE_FACTORS           	-	39	Variant:KP4 | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_39
SUBJECT_SAMPLE_FACTORS           	-	40	Variant:KP4 | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_40
SUBJECT_SAMPLE_FACTORS           	-	41	Variant:KP4 | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_41
SUBJECT_SAMPLE_FACTORS           	-	48	Variant:HPAC | Sample source:Pancreatic cancer cells	Replicate=1; RAW_FILE_NAME(Raw File name)=Cell_UCSF_48
SUBJECT_SAMPLE_FACTORS           	-	49	Variant:HPAC | Sample source:Pancreatic cancer cells	Replicate=2; RAW_FILE_NAME(Raw File name)=Cell_UCSF_49
SUBJECT_SAMPLE_FACTORS           	-	50	Variant:HPAC | Sample source:Pancreatic cancer cells	Replicate=3; RAW_FILE_NAME(Raw File name)=Cell_UCSF_50
SUBJECT_SAMPLE_FACTORS           	-	51	Variant:STD_1 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S02
SUBJECT_SAMPLE_FACTORS           	-	52	Variant:STD_2 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S03
SUBJECT_SAMPLE_FACTORS           	-	53	Variant:STD_3 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S04
SUBJECT_SAMPLE_FACTORS           	-	54	Variant:STD_4 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S05
SUBJECT_SAMPLE_FACTORS           	-	55	Variant:STD_5 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S06
SUBJECT_SAMPLE_FACTORS           	-	56	Variant:STD_6 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S07
SUBJECT_SAMPLE_FACTORS           	-	57	Variant:STD_7 | Sample source:STANDARD	Replicate=1; RAW_FILE_NAME(Raw File name)=00_STD_2x7DHC8t5_7DHDes4t5_S08
#COLLECTION
CO:COLLECTION_SUMMARY            	HPAC and KP4 pancreatic cancer cell lines were obtained from the American Type
CO:COLLECTION_SUMMARY            	Culture Collection (ATCC) or DSMZ. All cell lines were cultured in DMEM media
CO:COLLECTION_SUMMARY            	(Gibco) supplemented with 10% FBS (Atlanta biologicals), 1%
CO:COLLECTION_SUMMARY            	Pencillin/Streptomycin (Gibco) and 15mM HEPES (Gibco) and grown in a humidified
CO:COLLECTION_SUMMARY            	chamber at 37°C, 5% CO2. Cells were trypsinized using TrypLE (Gibco). Routine
CO:COLLECTION_SUMMARY            	mycoplasma testing was performed using the Mycoplasma PCR detection kit (Abm;
CO:COLLECTION_SUMMARY            	Cat. Number G238) at least once a month and the cell lines were authenticated by
CO:COLLECTION_SUMMARY            	STR fingerprinting. Cell lines are passaged for a maximum of 15 passages upon
CO:COLLECTION_SUMMARY            	thawing prior to replacement. To perform the experiments cells were washed with
CO:COLLECTION_SUMMARY            	PBS, detached and washed again two times with PBS before pellet freezing. KP4
CO:COLLECTION_SUMMARY            	WT, D374Y and R46L are expressing different human PCSK9 variants: WT, D374Y
CO:COLLECTION_SUMMARY            	(active variant), and R46L (inactive variant). KP4 control cells express a
CO:COLLECTION_SUMMARY            	control vector and KP4 WT express WT PCSK9. KP4 and HPAC are parental cells.
CO:SAMPLE_TYPE                   	Pancreatic cancer cells
#TREATMENT
TR:TREATMENT_SUMMARY             	Cell pellets were collected and suspended in 500 μL of
TR:TREATMENT_SUMMARY             	water-methanol-chloroform (1 :5 :2) in Eppendorf tubes. The samples were
TR:TREATMENT_SUMMARY             	homogenized on a MM 400 mill mixer with the aid of two 3-mm metal balls and at a
TR:TREATMENT_SUMMARY             	shaking frequency of 30Hz for 3min.The homogenization step was repeated two more
TR:TREATMENT_SUMMARY             	times. The samples were then ultra-sonicated in an ice-water bath for 2 min and
TR:TREATMENT_SUMMARY             	subsequently centrifuged at 21,000 g and 5°C for 10 min. The clear supernatants
TR:TREATMENT_SUMMARY             	were collected for the following LC-MS analysis and the precipitated pellets
TR:TREATMENT_SUMMARY             	were used for protein assay by UV-VIS spectroscopy using a standardized Bradford
TR:TREATMENT_SUMMARY             	procedure. No specific treatment was performed before the speficic mass
TR:TREATMENT_SUMMARY             	spectrometry processing protocol.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Cell pellets were collected and suspended in 500 μL of
SP:SAMPLEPREP_SUMMARY            	water-methanol-chloroform (1 :5 :2) in Eppendorf tubes. The samples were
SP:SAMPLEPREP_SUMMARY            	homogenized on a MM 400 mill mixer with the aid of two 3-mm metal balls and at a
SP:SAMPLEPREP_SUMMARY            	shaking frequency of 30Hz for 3min.The homogenization step was repeated two more
SP:SAMPLEPREP_SUMMARY            	times. The samples were then ultra-sonicated in an ice-water bath for 2 min and
SP:SAMPLEPREP_SUMMARY            	subsequently centrifuged at 21,000 g and 5°C for 10 min. The clear supernatants
SP:SAMPLEPREP_SUMMARY            	were collected for the following LC-MS analysis and the precipitated pellets
SP:SAMPLEPREP_SUMMARY            	were used for protein assay by UV-VIS spectroscopy using a standardized Bradford
SP:SAMPLEPREP_SUMMARY            	procedure.Specifically, Serially diluted standard solutions of
SP:SAMPLEPREP_SUMMARY            	7-dehydrocholesterol and 7-dehydrodesmosterol were prepared in acetonitrile. 
SP:SAMPLEPREP_SUMMARY            	The concentration range of each compound ranged from 0.00001µM to 1 µM with
SP:SAMPLEPREP_SUMMARY            	STD1 being the highest concentration and STD7 the lowest.   Next, 100 μL of
SP:SAMPLEPREP_SUMMARY            	the clear supernatant extractant from each sample was mixed with 300 μL of 50%
SP:SAMPLEPREP_SUMMARY            	methanol and 150 μL of chloroform. After vortex-mixing for 30 s and centrifugal
SP:SAMPLEPREP_SUMMARY            	clarification at 10,000 g for 2 min, the lower organic phase was collected and
SP:SAMPLEPREP_SUMMARY            	dried under a gentle nitrogen gas flow. The residue was dissolved in 100 µL of
SP:SAMPLEPREP_SUMMARY            	acetonitrile.  To 50 µL of the sample solution or each standard solution, 450
SP:SAMPLEPREP_SUMMARY            	µL of 10-mM 4-phenyl-1,2,4-triazoline-3,5-dione solution in acetonitrile was
SP:SAMPLEPREP_SUMMARY            	added. The mixtures were incubated at room temperature for 2 h on a Thermomixer
SP:SAMPLEPREP_SUMMARY            	at a shaking frequency of 600 rpm. 10 µL aliquots of the resultant solutions
SP:SAMPLEPREP_SUMMARY            	were injected into an Agilent Eclipse C18 (2.1*50 mm, 1.8 µm) column to run
SP:SAMPLEPREP_SUMMARY            	LC-MRM/MS on an Agilent 1290 UHPLC system coupled to a Sciex 7500 QQQ mass
SP:SAMPLEPREP_SUMMARY            	spectrometer operated in the positive-ion detection mode, using 0.1% formic acid
SP:SAMPLEPREP_SUMMARY            	in water (A) and 0.1% formic acid in acetonitrile (B) as the mobile phase for
SP:SAMPLEPREP_SUMMARY            	binary-solvent gradient elution under optimized LC separation and MRM/MS
SP:SAMPLEPREP_SUMMARY            	detection conditions
SP:SAMPLE_DERIVATIZATION         	Dansyl chloride
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Agilent Eclipse C18 (2.1*50 mm, 1.8 µm) column The gradient began at 65% B at 0
CH:CHROMATOGRAPHY_SUMMARY        	min, shifted to 50% B over 9 min, 100% B from 9 min to 9.1 min, then stayed at
CH:CHROMATOGRAPHY_SUMMARY        	100% B for 0.9 min before ramping down to 65% B in 0.1 min and stayed at 65% B
CH:CHROMATOGRAPHY_SUMMARY        	for 3.9 min, at 0.26 mL/min and 50 C.
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Agilent 1290
CH:COLUMN_NAME                   	Agilent ZORBAX Eclipse Plus C18 (50 x 2.1mm,1.8um)
CH:SOLVENT_A                     	100% water; 0.1% formic acid
CH:SOLVENT_B                     	66.67% isopropanol/33.33% acetonitrile; 0.1% formic acid
CH:FLOW_GRADIENT                 	The gradient began at 65% B at 0 min, shifted to 50% B over 9 min, 100% B from 9
CH:FLOW_GRADIENT                 	min to 9.1 min, then stayed at 100% B for 0.9 min before ramping down to 65% B
CH:FLOW_GRADIENT                 	in 0.1 min and stayed at 65% B for 3.9 min
CH:FLOW_RATE                     	0.26 mL/min
CH:COLUMN_TEMPERATURE            	50°C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	ABI Sciex 7500 QQQ
MS:INSTRUMENT_TYPE               	Triple quadrupole
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Concentrations of the compounds detected in the samples were calculated by
MS:MS_COMMENTS                   	interpolating their constructed linear-regression calibration curves with
MS:MS_COMMENTS                   	internal standard calibration or external standard calibration.
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	pmol/mg protein
MS_METABOLITE_DATA_START
Samples	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	39	40	41	48	49	50	51	52	53	54	55	56	57
Factors	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Variant:KP4_Control | Sample source:Pancreatic cancer cells	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Variant:KP4_D374Y | Sample source:Pancreatic cancer cells	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Variant:KP4_WT | Sample source:Pancreatic cancer cells	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Variant:KP4_R46L | Sample source:Pancreatic cancer cells	Variant:KP4 | Sample source:Pancreatic cancer cells	Variant:KP4 | Sample source:Pancreatic cancer cells	Variant:KP4 | Sample source:Pancreatic cancer cells	Variant:HPAC | Sample source:Pancreatic cancer cells	Variant:HPAC | Sample source:Pancreatic cancer cells	Variant:HPAC | Sample source:Pancreatic cancer cells	Variant:STD_1 | Sample source:STANDARD	Variant:STD_2 | Sample source:STANDARD	Variant:STD_3 | Sample source:STANDARD	Variant:STD_4 | Sample source:STANDARD	Variant:STD_5 | Sample source:STANDARD	Variant:STD_6 | Sample source:STANDARD	Variant:STD_7 | Sample source:STANDARD
7-Dehydrocholesterol	747.095569	571.024077	902.953631	1197.426395	1125.49759	7168.896312	9283.805793	3761.739927	2479.290345	2249.193819	375.41794	416.126907	4345.776136	4228.97696	5224.654874	579.850009	526.599358	3631.630486	2466.525991	2089.367108	63.552209	62.835299	51.540777	121.064303	92.550982	86.145937	N/A	N/A	N/A	N/A	N/A	N/A	N/A
7-Dehydrodesmosterol	34.254777	32.470984	6.215894	7.068508	7.068773	580.078633	682.3994	610.10392	28.091375	23.494704	78.009412	96.524283	254.327081	248.822096	332.314958	54.470758	54.373473	106.012093	35.495666	24.725216	8.551942	12.085504	9.968921	14.827474	14.613551	13.66286	N/A	N/A	N/A	N/A	N/A	N/A	N/A
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	Kegg_ID
7-Dehydrocholesterol	C04831
7-Dehydrodesmosterol	C15631
METABOLITES_END
#END