Summary of Study ST004018

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 PR002501. The data can be accessed directly via it's Project DOI: 10.21228/M8C55X 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 IDST004018
Study TitleEvaluation of SNAT1-dependent MeAIB uptake using GCMS
Study SummaryHY15549 cell lines were engineered to stably express a human SNAT1 cDNA or an empty vector (EV, controls) by lentiviral transduction. Either EV or +SNAT1 cell lines were plated and evaluated for MeAIB uptake by culturing cells with a gradient from 0.125-4 mM for 30-minutes in PBS for 30-minutes prior to GCMS analysis. SNAT1 exhibited saturable uptake kinetics with an apparent Km of ~2.91 mM.
Institute
University of British Columbia
DepartmentBiochemistry & Molecular Biology
LaboratoryParker laboratory
Last NameParker
First NameSeth
Address950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Emailseth.parker@bcchr.ca
Phone6048753121
Submit Date2025-06-19
Raw Data AvailableYes
Raw Data File Type(s)cdf
Analysis Type DetailGC-MS
Release Date2025-07-28
Release Version1
Seth Parker Seth Parker
https://dx.doi.org/10.21228/M8C55X
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR002501
Project DOI:doi: 10.21228/M8C55X
Project Title:Metabolomics analysis of SNAT2-deficient cells: implications for the discovery of selective transporter inhibitors
Project Type:Manuscript
Project Summary:Amino acid uptake by the solute carrier family of transporter proteins is critical to support cell metabolism, and inhibition of transporter activity represents a tractable strategy to restrict nutrient availability to cancer cells. A small molecule inhibitor of the sodium-coupled neutral amino acid transporter 2 (SNAT2), 3-(N-methyl(4-methylphenyl)sulfonamido)-N-(2-trifluoromethylbenzyl)thiophene-2-carboxamide (MMTC/57E), was recently identified and was shown to inhibit cell proliferation when combined with glucose transport inhibitors in breast and pancreatic cancer cell lines. In this study, we use mass spectrometry-based metabolomics and establish cell-based assays for the SNAT2 transporter. We show that SNAT2 knockout cells have significant defects in amino acid availability. Using our established assays, we fail to observe that MMTC/57E inhibits SNAT2 activity likely due to its poor solubility.
Institute:University of British Columbia
Department:Biochemistry & Molecular Biology
Laboratory:Parker laboratory
Last Name:Parker
First Name:Seth
Address:950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada
Email:seth.parker@bcchr.ca
Phone:6048753121

Subject:

Subject ID:SU004156
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Gender:Female

Factors:

Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Factor Doxycycline (y/n) Sample source MeAIB concentration (mM)
SA462043HYiSN2_EV_nodox_1aEV n HY15549 Pancreatic Cancer Cells 0.15625
SA462044HYiSN2_EV_nodox_1bEV n HY15549 Pancreatic Cancer Cells 0.15625
SA462045HYiSN2_EV_nodox_1cEV n HY15549 Pancreatic Cancer Cells 0.15625
SA462046HYiSN2_EV_nodox_2aEV n HY15549 Pancreatic Cancer Cells 0.3125
SA462047HYiSN2_EV_nodox_2cEV n HY15549 Pancreatic Cancer Cells 0.3125
SA462048HYiSN2_EV_nodox_2bEV n HY15549 Pancreatic Cancer Cells 0.3125
SA462049HYiSN2_EV_nodox_3cEV n HY15549 Pancreatic Cancer Cells 0.625
SA462050HYiSN2_EV_nodox_3bEV n HY15549 Pancreatic Cancer Cells 0.625
SA462051HYiSN2_EV_nodox_3aEV n HY15549 Pancreatic Cancer Cells 0.625
SA462052HYiSN2_EV_nodox_4cEV n HY15549 Pancreatic Cancer Cells 1.25
SA462053HYiSN2_EV_nodox_4aEV n HY15549 Pancreatic Cancer Cells 1.25
SA462054HYiSN2_EV_nodox_4bEV n HY15549 Pancreatic Cancer Cells 1.25
SA462055HYiSN2_EV_nodox_5cEV n HY15549 Pancreatic Cancer Cells 2.5
SA462056HYiSN2_EV_nodox_5bEV n HY15549 Pancreatic Cancer Cells 2.5
SA462057HYiSN2_EV_nodox_5aEV n HY15549 Pancreatic Cancer Cells 2.5
SA462058HYiSN2_EV_nodox_6bEV n HY15549 Pancreatic Cancer Cells 5
SA462059HYiSN2_EV_nodox_6aEV n HY15549 Pancreatic Cancer Cells 5
SA462060HYiSN2_EV_nodox_6cEV n HY15549 Pancreatic Cancer Cells 5
SA462061HYiSN2_EV_dox_1bEV y HY15549 Pancreatic Cancer Cells 0.15625
SA462062HYiSN2_EV_dox_1aEV y HY15549 Pancreatic Cancer Cells 0.15625
SA462063HYiSN2_EV_dox_2cEV y HY15549 Pancreatic Cancer Cells 0.3125
SA462064HYiSN2_EV_dox_2bEV y HY15549 Pancreatic Cancer Cells 0.3125
SA462065HYiSN2_EV_dox_2aEV y HY15549 Pancreatic Cancer Cells 0.3125
SA462066HYiSN2_EV_dox_3bEV y HY15549 Pancreatic Cancer Cells 0.625
SA462067HYiSN2_EV_dox_3cEV y HY15549 Pancreatic Cancer Cells 0.625
SA462068HYiSN2_EV_dox_3aEV y HY15549 Pancreatic Cancer Cells 0.625
SA462069HYiSN2_EV_dox_4cEV y HY15549 Pancreatic Cancer Cells 1.25
SA462070HYiSN2_EV_dox_4bEV y HY15549 Pancreatic Cancer Cells 1.25
SA462071HYiSN2_EV_dox_4aEV y HY15549 Pancreatic Cancer Cells 1.25
SA462072HYiSN2_EV_dox_5cEV y HY15549 Pancreatic Cancer Cells 2.5
SA462073HYiSN2_EV_dox_5bEV y HY15549 Pancreatic Cancer Cells 2.5
SA462074HYiSN2_EV_dox_5aEV y HY15549 Pancreatic Cancer Cells 2.5
SA462075HYiSN2_EV_dox_6cEV y HY15549 Pancreatic Cancer Cells 5
SA462076HYiSN2_EV_dox_6bEV y HY15549 Pancreatic Cancer Cells 5
SA462077HYiSN2_EV_dox_6aEV y HY15549 Pancreatic Cancer Cells 5
SA462078HYiSN2_SN1_nodox_1cSNAT1 n HY15549 Pancreatic Cancer Cells 0.15625
SA462079HYiSN2_SN1_nodox_1aSNAT1 n HY15549 Pancreatic Cancer Cells 0.15625
SA462080HYiSN2_SN1_nodox_1bSNAT1 n HY15549 Pancreatic Cancer Cells 0.15625
SA462081HYiSN2_SN1_nodox_2cSNAT1 n HY15549 Pancreatic Cancer Cells 0.3125
SA462082HYiSN2_SN1_nodox_2bSNAT1 n HY15549 Pancreatic Cancer Cells 0.3125
SA462083HYiSN2_SN1_nodox_2aSNAT1 n HY15549 Pancreatic Cancer Cells 0.3125
SA462084HYiSN2_SN1_nodox_3cSNAT1 n HY15549 Pancreatic Cancer Cells 0.625
SA462085HYiSN2_SN1_nodox_3bSNAT1 n HY15549 Pancreatic Cancer Cells 0.625
SA462086HYiSN2_SN1_nodox_3aSNAT1 n HY15549 Pancreatic Cancer Cells 0.625
SA462087HYiSN2_SN1_nodox_4cSNAT1 n HY15549 Pancreatic Cancer Cells 1.25
SA462088HYiSN2_SN1_nodox_4bSNAT1 n HY15549 Pancreatic Cancer Cells 1.25
SA462089HYiSN2_SN1_nodox_4aSNAT1 n HY15549 Pancreatic Cancer Cells 1.25
SA462090HYiSN2_SN1_nodox_5cSNAT1 n HY15549 Pancreatic Cancer Cells 2.5
SA462091HYiSN2_SN1_nodox_5bSNAT1 n HY15549 Pancreatic Cancer Cells 2.5
SA462092HYiSN2_SN1_nodox_5aSNAT1 n HY15549 Pancreatic Cancer Cells 2.5
SA462093HYiSN2_SN1_nodox_6aSNAT1 n HY15549 Pancreatic Cancer Cells 5
SA462094HYiSN2_SN1_nodox_6cSNAT1 n HY15549 Pancreatic Cancer Cells 5
SA462095HYiSN2_SN1_nodox_6bSNAT1 n HY15549 Pancreatic Cancer Cells 5
SA462096HYiSN2_SN1_dox_1bSNAT1 y HY15549 Pancreatic Cancer Cells 0.15625
SA462097HYiSN2_SN1_dox_1aSNAT1 y HY15549 Pancreatic Cancer Cells 0.15625
SA462098HYiSN2_SN1_dox_1cSNAT1 y HY15549 Pancreatic Cancer Cells 0.15625
SA462099HYiSN2_SN1_dox_2aSNAT1 y HY15549 Pancreatic Cancer Cells 0.3125
SA462100HYiSN2_SN1_dox_2bSNAT1 y HY15549 Pancreatic Cancer Cells 0.3125
SA462101HYiSN2_SN1_dox_2cSNAT1 y HY15549 Pancreatic Cancer Cells 0.3125
SA462102HYiSN2_SN1_dox_3cSNAT1 y HY15549 Pancreatic Cancer Cells 0.625
SA462103HYiSN2_SN1_dox_3bSNAT1 y HY15549 Pancreatic Cancer Cells 0.625
SA462104HYiSN2_SN1_dox_3aSNAT1 y HY15549 Pancreatic Cancer Cells 0.625
SA462105HYiSN2_SN1_dox_4cSNAT1 y HY15549 Pancreatic Cancer Cells 1.25
SA462106HYiSN2_SN1_dox_4aSNAT1 y HY15549 Pancreatic Cancer Cells 1.25
SA462107HYiSN2_SN1_dox_4bSNAT1 y HY15549 Pancreatic Cancer Cells 1.25
SA462108HYiSN2_SN1_dox_5aSNAT1 y HY15549 Pancreatic Cancer Cells 2.5
SA462109HYiSN2_SN1_dox_5bSNAT1 y HY15549 Pancreatic Cancer Cells 2.5
SA462110HYiSN2_SN1_dox_5cSNAT1 y HY15549 Pancreatic Cancer Cells 2.5
SA462111HYiSN2_SN1_dox_6bSNAT1 y HY15549 Pancreatic Cancer Cells 5
SA462112HYiSN2_SN1_dox_6aSNAT1 y HY15549 Pancreatic Cancer Cells 5
SA462113HYiSN2_SN1_dox_6cSNAT1 y HY15549 Pancreatic Cancer Cells 5
Showing results 1 to 71 of 71

Collection:

Collection ID:CO004149
Collection Summary:Cells were washed twice with room temperature (20°C) PBS before adding 2 mL of PBS containing 0.16-5 mM of MeAIB. After a 30-minute incubation at 20°C, the transport buffer was aspirated and the cells were washed three times with ice cold 0.9% NaCl. To extract metabolites, 1 mL of ice cold 80% methanol containing 2.5 nmol of 13C3-alanine was added to each well. The plates were shaken at ~500 rpm for 15 minutes at room temperature and 900 µL was transferred to a new tube and dried using a SpeedVac for GCMS analysis.
Sample Type:Tumor cells

Treatment:

Treatment ID:TR004165
Treatment Summary:No treatment.

Sample Preparation:

Sampleprep ID:SP004162
Sampleprep Summary:Dried samples were derivatized for GCMS analysis. 25 µL of methoxyamine hydrochloride (20 mg/mL, prepared in pyridine) was added to each tube, which was vortexed and incubated at 37°C for 60 minutes. Following the incubation, 25 µL of MTBSTFA + 1% TBDMSCl was added to each tube, which was vortexed and centrifuged at 21,300 x g for 15-minutes at 20°C. 40 µL of the resulting supernatant was transferred to a plastic sample vial for GCMS analysis.

Chromatography:

Chromatography ID:CH005033
Chromatography Summary:Agilent DB35-MS (30m x 0.25mm, 0.25µm) was interfaced with an inert column (5m x 0.15mm, 0 µm) to the MSD. Initial oven temperature maintained at 75°C for 3 minutes, ramped to 255°C at 7.5°C/min, held for 10 minutes, ramped to 320°C at 10°C/min, held for 5 minutes. The inlet was maintained at 270°C in splitless mode; an injection volume of 1 µL was used for all samples.
Instrument Name:Agilent 8890
Column Name:Agilent DB-35MS (30m x 0.25mm, 0.25µm) interfaced with an inert column (5m x 0.15mm, 0 µm)
Column Temperature:75°C
Flow Gradient:100% Helium
Flow Rate:1 mL/min
Solvent A:UHP Helium
Solvent B:None
Chromatography Type:GC

Analysis:

Analysis ID:AN006624
Analysis Type:MS
Chromatography ID:CH005033
Num Factors:24
Num Metabolites:4
Units:ion counts
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