Summary of Study ST003123

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 PR001941. The data can be accessed directly via it's Project DOI: 10.21228/M8TM76 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.

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Study IDST003123
Study TitleOMM1.3 uveal melanoma cells fed with [U-13C6] glucose and treated with Fasnall for 4 h
Study TypeIntracellular metabolomics, [U-13C6] glucose tracing
Study SummaryOMM1.3 uveal melanoma cells fed with [U-13C6] glucose and treated with Fasnall for 4 h. Cells were grown in RPMI-1640 with 10% dialyzed FBS.
Institute
Wistar Institute
DepartmentMolecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
LaboratorySchug's Lab
Last NameMukha
First NameDzmitry
Address3601 Spruce St, Philadelphia, PA 19104, USA
Emaildmukha@wistar.org
Phone2154956903
Submit Date2023-12-11
Num Groups7
Total Subjects21
PublicationsTBD
Raw Data AvailableYes
Raw Data File Type(s)mzXML, raw(Thermo)
Analysis Type DetailLC-MS
Release Date2024-03-29
Release Version1
Dzmitry Mukha Dzmitry Mukha
https://dx.doi.org/10.21228/M8TM76
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR001941
Project DOI:doi: 10.21228/M8TM76
Project Title:The shutdown of NADH oxidation via Respiratory Complex I mimics fatty acid biosynthesis inhibition
Project Type:LC-MS Quantitative Analysis
Project Summary:Proliferating cancer cells actively utilize anabolic processes for biomass production, including de novo biosynthesis of amino acids, nucleotides, and fatty acids. The key enzyme of the fatty acid biosynthesis pathway, fatty acid synthase (FASN), is widely recognized as a promising therapeutic target in cancer and other health conditions. Here, we establish a metabolic signature of FASN inhibition using a panel of pharmacological inhibitors (GSK2194069, TVB-2640, TVB-3166, C75, cerulenin, and Fasnall). We find that the activity of some commonly used FASN inhibitors is inconsistent with the metabolic signature of FASN inhibition (accumulation of malonate, succinate, malonyl coenzyme A, succinyl coenzyme A, and other metabolic perturbations). Moreover, we show that one of these putative FASN inhibitors, Fasnall, is a respiratory Complex I inhibitor that mimics FASN inhibition through NADH accumulation and consequent depletion of the tricarboxylic acid cycle metabolites. We demonstrate that Fasnall impairs tumor growth in several oxidative phosphorylation-dependent cancer models, including combination therapy-resistant melanoma patient-derived xenografts. Fasnall administration does not reproduce neurological side effects in mice reported for other Complex I inhibitors. Our results have significant implications for understanding the FASN role in human health and disease and provide evidence of therapeutic potential for Complex I inhibitors with fast systemic clearance. Our findings also highlight the continuing need for validation of small molecule inhibitors to distinguish high-quality chemical probes and to expand the understanding of their application.
Institute:Wistar Institute
Department:Molecular and Cellular Oncogenesis Program, Ellen and Ronald Caplan Cancer Center
Laboratory:Schug's Lab
Last Name:Mukha
First Name:Dzmitry
Address:3601 Spruce St., Philadelphia, Pennsylvania 19104, USA
Email:dmukha@wistar.org
Phone:+12154956903
Funding Source:This work was supported by grants from the National Institutes of Health (NIH) National Cancer Institute (NCI) DP2 CA249950-01 (Z.T.S.), NIH NCI P01 CA114046 (Z.T.S.), Melanoma Research Foundation 717173 (Z.T.S.), and Susan G. Komen CCR19608782 (Z.T.S.).
Publications:Submission Pending
Contributors:Dzmitry Mukha, Jena Dessain, Seamus O’Connor, Katherine Pniewski, Fabrizio Bertolazzi, Jeet Patel, Mary Mullins, Zachary T. Schug

Subject:

Subject ID:SU003240
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Genotype Strain:626A>C mutation in GNAQ resulting in Q209P (PMC3288394)
Age Or Age Range:NA
Weight Or Weight Range:NA
Height Or Height Range:NA
Cell Biosource Or Supplier:Dr. Andrew Aplin's lab, Thomas Jefferson University (Philadelphia, PA, USA)
Cell Strain Details:OMM1.3, uveal melanoma, derived from a metastatic lesion
Subject Comments:The cell line is also known as OMM2.3
Cell Primary Immortalized:No
Cell Passage Number:NA
Species Group:Mammals

Factors:

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

mb_sample_id local_sample_id Type of glucose Fasnall Concentration
SA338137Blank-80MeOH_20211219013554NA NA
SA338138Blank-80MeOH_20211218154318NA NA
SA338139m21-L327_SO_05U-13C6 0 nM
SA338140m21-L327_SO_06U-13C6 0 nM
SA338141m21-L327_SO_04U-13C6 0 nM
SA338145m21-L327_SO_11U-13C6 100 nM
SA338146m21-L327_SO_12U-13C6 100 nM
SA338147m21-L327_SO_10U-13C6 100 nM
SA338142m21-L327_SO_17U-13C6 1 uM
SA338143m21-L327_SO_16U-13C6 1 uM
SA338144m21-L327_SO_18U-13C6 1 uM
SA338154m21-L327_SO_13U-13C6 500 nM
SA338155m21-L327_SO_15U-13C6 500 nM
SA338156m21-L327_SO_14U-13C6 500 nM
SA338151m21-L327_SO_09U-13C6 50 nM
SA338152m21-L327_SO_07U-13C6 50 nM
SA338153m21-L327_SO_08U-13C6 50 nM
SA338148m21-L327_SO_19U-13C6 5 uM
SA338149m21-L327_SO_20U-13C6 5 uM
SA338150m21-L327_SO_21U-13C6 5 uM
SA338157m21-L327_SO_01unlabeled 0 nM
SA338158m21-L327_SO_03unlabeled 0 nM
SA338159m21-L327_SO_02unlabeled 0 nM
Showing results 1 to 23 of 23

Collection:

Collection ID:CO003233
Collection Summary:One PBS washing, 80% methanol extraction.
Collection Protocol Filename:DM_metabolomics_samples.txt
Sample Type:Cultured cells
Collection Method:80% methanol extraction
Volumeoramount Collected:500 ul
Storage Conditions:-80℃
Collection Vials:1.7 ml plastic centrifuge tubes
Storage Vials:1.7 ml plastic centrifuge tubes

Treatment:

Treatment ID:TR003249
Treatment Summary:Cells were grown in RPMI-1640 supplemented with [U-13C6] glucose and treated with various Fasnall concentrations.
Treatment Compound:Fasnall
Treatment Dose:50 nM - 5 uM
Treatment Doseduration:4 h
Treatment Vehicle:DMSO
Cell Growth Container:6-well plates
Cell Media:RPMI-1640
Cell Envir Cond:37C, 5% CO2
Cell Pct Confluence:~70%

Sample Preparation:

Sampleprep ID:SP003247
Sampleprep Summary:Metabolites were extracted with ice-cold 80% methanol.
Sampleprep Protocol Filename:DM_metabolomics_samples.txt
Extraction Method:80% methanol
Extract Enrichment:None
Extract Cleanup:None
Extract Storage:-80℃
Sample Resuspension:None
Sample Derivatization:None
Sample Spiking:None
Subcellular Location:Intracellular metabolites

Chromatography:

Chromatography ID:CH003872
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um)
Column Temperature:40
Flow Gradient:0-12.5 min, 80-30% B; 12.5-15 min, 30% B; 15-15.2 min, 30-80% B; 15.2-22.5 min, 80% B
Flow Rate:0-20 min, 0.2 ml/min; 20-21 min 0.2-0.3 ml/min; 21-22 min, 0.3 ml/min; 22-22.1 min, 0.2 ml/min; 22.1-22.5 min, 0.2 ml/min
Solvent A:100% water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate
Solvent B:100% acetonitrile
Chromatography Type:HILIC

Analysis:

Analysis ID:AN005117
Laboratory Name:Wistar Institute Proteomics & Metabolomic Core
Analysis Type:MS
Operator Name:Nicole Gorman
Detector Type:Orbitrap
Chromatography ID:CH003872
Num Factors:8
Num Metabolites:189
Rt Units:Minutes
Units:AU
  
Analysis ID:AN005118
Laboratory Name:Wistar Institute Proteomics & Metabolomic Core
Analysis Type:MS
Operator Name:Nicole Gorman
Detector Type:Orbitrap
Chromatography ID:CH003872
Num Factors:8
Num Metabolites:206
Rt Units:Minutes
Units:AU
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