Summary of Study ST003152
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.
| Study ID | ST003152 |
| Study Title | Metabolomics analysis of murine xenograft tumors derived from human breast cancer cell line MCF7 1 h after isotopic glucose bolus |
| Study Type | Intracellular metabolomics, [U-13C6] glucose tracing |
| Study Summary | Metabolomics analysis of murine xenograft tumors derived from human breast cancer cell line MCF7 1 h after isotopic glucose bolus. The NSG female mice were ~55 days after grafting tumors. [U-13C6] D-Glucose bolus was delivered intraperitoneally with a 10 mg/kg dose of Fasnall. |
| 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, PA 19104, USA |
| dmukha@wistar.org | |
| Phone | 2154956903 |
| Submit Date | 2024-03-09 |
| Num Groups | 2 |
| Total Subjects | 12 |
| Num Females | 12 |
| Publications | Submission Pending |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2024-04-02 |
| Release Version | 1 |
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: | SU003269 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | NOD Scid Gamma (NSG) |
| Age Or Age Range: | ~4.5 months |
| Weight Or Weight Range: | 24.3 +/- 1.9 g (mean +/- SD) |
| Gender: | Female |
| Animal Housing: | Wistar Animal Facility |
| Species Group: | Rodents (order Rodentia) |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Isotopic Tracer | Drug Treatment |
|---|---|---|---|---|
| SA340980 | 80MeOH_Blank_20230918155328 | NA | NA | NA |
| SA340981 | 80MeOH_Blank_20230918220912 | NA | NA | NA |
| SA340982 | 135standard_091823_1 | NA | NA | NA |
| SA340983 | 135standard_091823_2 | NA | NA | NA |
| SA340984 | 80MeOH_Blank | NA | NA | NA |
| SA340985 | 80MeOH_Blank_20230918223604 | NA | NA | NA |
| SA340986 | 80MeOH_Blank_20230918145946 | NA | NA | NA |
| SA340987 | 04_Fasnall_MCF7_Tum_Gluc_1h_F_1070 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340988 | 03_Fasnall_MCF7_Tum_Gluc_1h_F_1069 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340989 | 05_Fasnall_MCF7_Tum_Gluc_1h_F_1071 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340990 | 06_Fasnall_MCF7_Tum_Gluc_1h_F_1072 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340991 | 01_Fasnall_MCF7_Tum_Gluc_1h_F_1066 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340992 | 07_Fasnall_MCF7_Tum_Gluc_1h_F_1073 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340993 | 02_Fasnall_MCF7_Tum_Gluc_1h_F_1068 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | 10 mg/kg Fasnall |
| SA340994 | 10_Fasnall_MCF7_Tum_Gluc_1h_D_1077 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | Vehicle |
| SA340995 | 08_Fasnall_MCF7_Tum_Gluc_1h_D_1074 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | Vehicle |
| SA340996 | 09_Fasnall_MCF7_Tum_Gluc_1h_D_1075 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | Vehicle |
| SA340997 | 11_Fasnall_MCF7_Tum_Gluc_1h_D_1079 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | Vehicle |
| SA340998 | 12_Fasnall_MCF7_Tum_Gluc_1h_D_1080 | Xenograft tumors | 1 g/kg [U-13C6] D-glucose in PBS | Vehicle |
| Showing results 1 to 19 of 19 |
Collection:
| Collection ID: | CO003262 |
| Collection Summary: | Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. Each sample was weighed while frozen. Frozen samples were ground at the temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added to each tube at a ratio of 1 ml per 50 mg of tissue. |
| Collection Protocol Filename: | DM_metabolomics_samples.txt |
| Sample Type: | Tumor cells |
| Collection Method: | 80% methanol extraction |
| Volumeoramount Collected: | ~1 ml |
| Storage Conditions: | -80℃ |
| Collection Vials: | 1.5 ml plastic centrifuge tubes |
| Storage Vials: | 1.5 ml plastic centrifuge tubes |
Treatment:
| Treatment ID: | TR003278 |
| Treatment Summary: | Animals received 1 g/kg [U-13C6] D-glucose bolus intraperitoneally with a 10 mg/kg dose of Fasnall. |
| Treatment Compound: | Fasnall |
| Treatment Route: | Intraperitoneal injection |
| Treatment Dose: | 10 mg/kg |
| Treatment Dosevolume: | 50 ul |
| Treatment Doseduration: | 1 h |
| Treatment Vehicle: | 1:1 DMSO/PBS |
| Animal Anesthesia: | None |
| Animal Fasting: | None |
| Animal Endp Euthanasia: | CO2 |
| Animal Endp Tissue Coll List: | 50 mg tumor samples |
Sample Preparation:
| Sampleprep ID: | SP003276 |
| Sampleprep Summary: | Tissue metabolite extracts were prepared from ~50 mg snap-frozen tissue samples. Each sample was weighed while frozen. Frozen samples were ground at the temperature of liquid nitrogen by Retch Cryomill. Then, 80% methanol was added to each tube at a ratio of 1 ml per 50 mg of tissue. All samples were centrifuged at 18,000 g and 4 °C for 20 min. The supernatant was then transferred to new Eppendorf tubes and centrifuged again with the same parameters. After centrifugation, the extracts were transferred to glass LC-MS vials. |
| 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 |
Combined analysis:
| Analysis ID | AN005171 | AN005172 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
| Column | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | AU | AU |
Chromatography:
| Chromatography ID: | CH003912 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 40 |
| Flow Gradient: | A linear solvent gradient of a total duration 22.5 min was starting with 0.2 ml/min flow rate of 80% solvent B, 12.5 min – 30%, 15 min – 30%, 15.2 min – 80%, 20 min – 80%, 21 min – flow rate 0.3 ml/min, 22 min – flow rate 0.3 ml/min, 22.1 min – flow rate 0.2 ml/min. |
| Flow Rate: | 0.2 ml/min |
| Solvent A: | 100% Water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004906 |
| Analysis ID: | AN005171 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN. |
| Ion Mode: | NEGATIVE |
| Ion Source Temperature: | 200 |
| Ion Spray Voltage: | 3600 |
| Ionization: | Both |
| Source Temperature: | 200 |
| Automatic Gain Control: | 5e6 |
| Dataformat: | RAW |
| MS ID: | MS004907 |
| Analysis ID: | AN005172 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN. |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 200 |
| Ion Spray Voltage: | 3600 |
| Ionization: | Both |
| Source Temperature: | 200 |
| Automatic Gain Control: | 5e6 |
| Dataformat: | RAW |
