Summary of Study ST003734

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002320. The data can be accessed directly via it's Project DOI: 10.21228/M8R83M This work is supported by NIH grant, U2C- DK119886.

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This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST003734
Study TitleAltered Omega-6/Omega-3 PUFA Ratios and Phospholipid Profiles in CFTR-Mutant PANC-1 Cells Reveal Novel Links Between CFTR Function and Lipid Metabolism
Study SummaryCFTR mutations have been implicated in pancreatitis and pancreatic cancer. CFTR is known to affect lipid metabolism and immune regulation. In this study, we employed untargeted lipidomics with iterative tandem mass spectrometry (MS/MS) to characterize the differences in PANC-1 cells with different CFTR states (overexpression, empty vector, and mutations). Our study found that three PANC-1 cells carrying CFTR mutations (p.L88X, p.E681V, p.C1355Y) showed a significantly higher ratio of FA 20:4 (omega-6 PUFA) and FA 22:6 (omega-3 PUFA) compared to the CFTR wild-type (CFTR OE) PANC-1 cells. Among the arachidonoyl species, the compounds PC16:0_20:4 and PC18:0_20:4 showed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PS18:0_20:4 exhibited a difference only between CFTR p.L88X PANC-1 cells and CFTR OE. The compounds PI16:0_20:4 and PI18:0_20:4 showed differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE. In the docosahexaenoyl species, the compounds PC18:0_22:6 and PI18:0_22:6 displayed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PC16:0_22:6 exhibited differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE.
Institute
Changhai Hospital
Last NameJin
First NameGang
Address168 Changhai Road, Yangpu District, Shanghai, China, 200433
Emailjingang@smmu.edu.cn
Phone+86 18621670329
Submit Date2025-02-10
Raw Data AvailableYes
Raw Data File Type(s)mzML, raw(Thermo)
Analysis Type DetailLC-MS
Release Date2025-02-19
Release Version1
Gang Jin Gang Jin
https://dx.doi.org/10.21228/M8R83M
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002320
Project DOI:doi: 10.21228/M8R83M
Project Title:CFTR Mutations Regulate Lipid Homeostasis and Inflammatory Signaling in Pancreatic Cancer
Project Summary:CFTR mutations have been implicated in pancreatitis and pancreatic cancer. CFTR is known to affect lipid metabolism and immune regulation. In this study, we employed untargeted lipidomics with iterative tandem mass spectrometry (MS/MS) to characterize the differences in PANC-1 cells with different CFTR states (overexpression, empty vector, and mutations). Our study found that three PANC-1 cells carrying CFTR mutations (p.L88X, p.E681V, p.C1355Y) showed a significantly higher ratio of FA 20:4 (omega-6 PUFA) and FA 22:6 (omega-3 PUFA) compared to the CFTR wild-type (CFTR OE) PANC-1 cells. Among the arachidonoyl species, the compounds PC16:0_20:4 and PC18:0_20:4 showed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PS18:0_20:4 exhibited a difference only between CFTR p.L88X PANC-1 cells and CFTR OE. The compounds PI16:0_20:4 and PI18:0_20:4 showed differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE. In the docosahexaenoyl species, the compounds PC18:0_22:6 and PI18:0_22:6 displayed higher normalized intensities in CFTR-mutated PANC-1 cells compared to CFTR OE. The compound PC16:0_22:6 exhibited differences comparing CFTR p.L88X and p.C1355Y cells with CFTR OE.
Institute:Changhai Hospital
Last Name:Jin
First Name:Gang
Address:168 Changhai Road, Yangpu District, Shanghai, China, 200433
Email:jingang@smmu.edu.cn
Phone:+86 18621670329

Subject:

Subject ID:SU003866
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id CFTR state
SA407476NEG_C1355Y_2C1355Y
SA407477POS_C1355Y_1C1355Y
SA407478POS_C1355Y_2C1355Y
SA407479NEG_C1355Y_4C1355Y
SA407480NEG_C1355Y_3C1355Y
SA407481NEG_C1355Y_1C1355Y
SA407482POS_C1355Y_3C1355Y
SA407483POS_C1355Y_4C1355Y
SA407484POS_E681V_1E681V
SA407485NEG_E681V_1E681V
SA407486NEG_E681V_2E681V
SA407487NEG_E681V_3E681V
SA407488NEG_E681V_4E681V
SA407489POS_E681V_2E681V
SA407490POS_E681V_3E681V
SA407491POS_E681V_4E681V
SA407492POS_EV_2EV
SA407493POS_EV_1EV
SA407494NEG_EV_4EV
SA407495NEG_EV_1EV
SA407496NEG_EV_2EV
SA407497NEG_EV_3EV
SA407498POS_EV_4EV
SA407499POS_EV_3EV
SA407500NEG_L88X_1L88X
SA407501NEG_L88X_4L88X
SA407502POS_L88X_4L88X
SA407503POS_L88X_3L88X
SA407504POS_L88X_2L88X
SA407505POS_L88X_1L88X
SA407506NEG_L88X_3L88X
SA407507NEG_L88X_2L88X
SA407508NEG_OE_1OE
SA407509POS_OE_4OE
SA407510POS_OE_3OE
SA407511NEG_OE_3OE
SA407512NEG_OE_4OE
SA407513POS_OE_1OE
SA407514POS_OE_2OE
SA407515NEG_OE_2OE
SA407516NEG_QC_5QC
SA407517NEG_QC_4QC
SA407518NEG_QC_3QC
SA407519NEG_QC_2QC
SA407520NEG_QC_6QC
SA407521POS_QC_1QC
SA407522POS_QC_2QC
SA407523POS_QC_3QC
SA407524POS_QC_4QC
SA407525POS_QC_5QC
SA407526POS_QC_6QC
SA407527NEG_QC_1QC
Showing results 1 to 52 of 52

Collection:

Collection ID:CO003859
Collection Summary:The human pancreatic cancer cell line PANC-1 was obtained from the National Collection of Authenticated Cell Cultures.Cells were cultured in DMEM medium supplemented with 10% FBS and 1% penicillin/streptomycin at 37°C with 5% CO2. Cells were grown to 80-90% confluence in 10-cm dishes. For sample collection, cells were washed twice with ice-cold PBS, harvested by trypsinization, and collected by centrifugation at 1000g for 5 minutes at 4°C. The cell pellets were immediately processed for metabolite extraction.
Sample Type:Pancreas

Treatment:

Treatment ID:TR003875
Treatment Summary:We used lentiviral vectors (OBiO Technology, Shanghai, China) to construct various recombinant vectors for overexpressing CFTR and its mutant variants. The vectors included CFTR overexpressing vector (CFTR OE), CFTR p.L88X overexpressing vector (CFTR p.L88X mt), CFTR p.E681V overexpressing vector (CFTR p.E681V mt),CFTR p.C1355Y overexpressing vector (CFTR p.C1355Y mt), and an empty plasmid serving as a negative control (EV).To introduce these vectors into PANC-1 cells, we infected cells at 40%‐50% confluence with lentiviral vectors at an appropriate multiplicity of infection (MOI). Following infection, stably transfected cells were selected using puromycin according to standard protocols.PANC-1 cell line (OE, EV, L88X, E681V, C1355Y) was set up with 4 technical replicates.

Sample Preparation:

Sampleprep ID:SP003872
Sampleprep Summary:The lipid extractions from PANC-1 cells (CFTR EV, OE, p.L88X, p.E681V, and p.C1355Y) were dried in a lyophilizer and then the lipids were resuspended in 100 μL isopropanol and methanal (volume 1:1). Waters ACQUITY UPLC coupled to Obritrap-LTQ-Elite (Thermo Fisher, USA) was used for lipidomic analyses. A Waters ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 μm) was used for lipid separation. The mobile phases consisted of 10 mM ammonium formate and 0.1% formic acid dissolved in 6:4 (v/v) ACN/H2O (phase A) and 9:1 (v/v) isopropanol/ACN (phase B). The flow rate was set at 0.3 mL/min, the injection volume was 4 μL, and the column temperature was at 45 °C. The elution gradient: 0-2 minutes, 70%-57% A; 2-2.1 minutes, 57%-45% A; 2.1-12 minutes, 45%-35% A; 12-18 minutes, 35%-15% A; 18-20 minutes, 15%-0% A; 20-25 minutes, 0%-70% A.

Chromatography:

Chromatography ID:CH004652
Instrument Name:Waters Acquity
Column Name:Waters ACQUITY UPLC BEH C18 (100 x 2.1 mm, 1.7 μm)
Column Temperature:40°C
Flow Gradient:0-2 minutes, 70%-57% A; 2-2.1 minutes, 57%-45% A; 2.1-12 minutes, 45%-35% A; 12-18 minutes, 35%-15% A; 18-20 minutes, 15%-0% A; 20-25 minutes, 0%-70% A.
Flow Rate:0.3 mL/min
Solvent A:60% acetonitrile/40% Water; 10mM ammonium formate; 0.1% formic acid
Solvent B:90% isopropanol/10% acetonitrile; 10mM ammonium formate; 0.1% formic acid
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN006126
Analysis Type:MS
Chromatography ID:CH004652
Num Factors:6
Num Metabolites:27
Rt Units:Minutes
Units:Peak area
  
Analysis ID:AN006127
Analysis Type:MS
Chromatography ID:CH004652
Num Factors:6
Num Metabolites:516
Rt Units:Minutes
Units:Peak area
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