Summary of Study ST004022
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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
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.
| Study ID | ST004022 |
| Study Title | Evaluating competitive inhibition of SNAT2-dependent MeAIB uptake by SNAT2 knockout or treatment with L-alanine or 57E. |
| Study Summary | MeAIB is a competitive inhibitor and transported substrate of SNAT2. In this study, we quantified cellular uptake of MeAIB and how L-alanine or 57E treatment might inhibit its uptake in HY15549 cells. |
| 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 |
| seth.parker@bcchr.ca | |
| Phone | 6048753121 |
| Submit Date | 2025-06-19 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf |
| Analysis Type Detail | GC-MS |
| Release Date | 2025-07-28 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
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: | SU004160 |
| 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 | Treatment | Doxycycline (y/n) | Sample source | Final DMSO (%) | MeAIB concentration (mM) |
|---|---|---|---|---|---|---|
| SA462546 | HY15549_iSNAT2_wDox_57E20_M500_r2 | 57E (20 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462547 | wDox_57E20_M500_1 | 57E (20 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462548 | HY15549_iSNAT2_wDox_57E20_M500_r3 | 57E (20 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462549 | HY15549_iSNAT2_5DMSO_wDox_57E20_M500_4 | 57E (20 microM) | yes | HY15549 Pancreatic Cancer Cells | 5 | 0.5 |
| SA462550 | HY15549_iSNAT2_wDox_57E40_M500_r2 | 57E (40 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462551 | HY15549_iSNAT2_wDox_57E40_M500_r3 | 57E (40 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462552 | wDox_57E40_M500_1 | 57E (40 microM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462553 | HY15549_iSNAT2_wDox_Ala20_M500_r3 | L-Alanine (20 mM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462554 | HY15549_iSNAT2_wDox_Ala20_M500_r2 | L-Alanine (20 mM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462555 | wDox_Ala20mM_M500_1 | L-Alanine (20 mM) | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462556 | HY15549_iSNAT2_5DMSO_wDox_Ala20_M500_4 | L-Alanine (20 mM) | yes | HY15549 Pancreatic Cancer Cells | 5 | 0.5 |
| SA462557 | noDox_veh_M500_1 | vehicle | no | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462558 | noDox_veh_M500_2 | vehicle | no | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462559 | HY15549_iSNAT2_noDox_veh_M500_r3 | vehicle | no | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462560 | HY15549_iSNAT2_noDox_veh_M500_r2 | vehicle | no | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462561 | HY15549_iSNAT2_5DMSO_noDox_veh_M500_4 | vehicle | no | HY15549 Pancreatic Cancer Cells | 5 | 0.5 |
| SA462562 | HY15549_iSNAT2_wDox_veh_M500_r3 | vehicle | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462563 | HY15549_iSNAT2_wDox_veh_M500_r2 | vehicle | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462564 | wDox_veh_M500_1 | vehicle | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462565 | wDox_veh_M500_2 | vehicle | yes | HY15549 Pancreatic Cancer Cells | 0.1 | 0.5 |
| SA462566 | HY15549_iSNAT2_5DMSO_wDox_veh_M500_4 | vehicle | yes | HY15549 Pancreatic Cancer Cells | 5 | 0.5 |
| Showing results 1 to 21 of 21 |
Collection:
| Collection ID: | CO004153 |
| Collection Summary: | Cells were washed twice with room temperature (20°C) PBS before adding 2 mL of PBS containing 0.5 mM MeAIB and 20 mM of L-alanine, 20 µM of 57E, or 40 µM of 57E. A separate well was plated without doxycycline for 24-hours to withdraw SNAT2 expression (SNAT2-KO). 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 nM 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: | TR004169 |
| Treatment Summary: | No treatment. |
Sample Preparation:
| Sampleprep ID: | SP004166 |
| 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: | CH005037 |
| Chromatography Summary: | Agilent DB35-MS (30 m x 0.25 mm, 0.25 µm) was interfaced with an inert column (5 m x 0.15 mm, 0 µm) to the MSD. Initial oven temperature maintained at 75°C for 3 minutes, ramped to 255°C at 7.5C/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: | AN006628 |
| Analysis Type: | MS |
| Chromatography ID: | CH005037 |
| Num Factors: | 9 |
| Num Metabolites: | 4 |
| Units: | ion counts |
