Summary of Study ST003434

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.

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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 IDST003434
Study TitlePlasma concentrations of Fasnall in mice after a bolus of 10 mg/kg administered intraperitoneally.
Study TypePlasma metabolomics
Study SummaryA single dose of 10 mg/kg (drug/body weight) Fasnall was injected intraperitoneally (~50 ul of solution in 1:1 DMSO:PBS), and plasma samples were collected. Blood was collected through a cardiac puncture. EDTA was used as an anticoagulant.
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
Phone+12154956903
Submit Date2024-08-21
Num Groups6
Total Subjects19
PublicationsSubmission Pending
Raw Data AvailableYes
Raw Data File Type(s)mzML, wiff
Analysis Type DetailLC-MS
Release Date2024-09-12
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:SU003561
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090
Age Or Age Range:3 months
Weight Or Weight Range:24-33 g
Gender:Female
Animal Animal Supplier:Wistar Animal Facility
Animal Housing:Wistar Animal Facility
Animal Feed:Standard chow diet

Factors:

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

mb_sample_id local_sample_id Sample source Sample Type Drug Treatment
SA37805909 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1084BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806003 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1081BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806114 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1087BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806205 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1082BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806313 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1086BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806407 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1083BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806511 BT-474 Xeno D3 Acetate 6 hrs Plasma Fasnall 1085BT-474 Xenograft in an NSG mouse Tumor 10 mg/kg Fasnall, 2 g/kg [D3] acetate bolus
SA37806612 BT-474 Xeno D3 Acetate 6 hrs Plasma DMSO 1093BT-474 Xenograft in an NSG mouse Tumor Vehicle (DMSO), 2 g/kg [D3] acetate bolus
SA37806710 BT-474 Xeno D3 Acetate 6 hrs Plasma DMSO 1092BT-474 Xenograft in an NSG mouse Tumor Vehicle (DMSO), 2 g/kg [D3] acetate bolus
SA37806808 BT-474 Xeno D3 Acetate 6 hrs Plasma DMSO 1091BT-474 Xenograft in an NSG mouse Tumor Vehicle (DMSO), 2 g/kg [D3] acetate bolus
SA37806906 BT-474 Xeno D3 Acetate 6 hrs Plasma DMSO 1090BT-474 Xenograft in an NSG mouse Tumor Vehicle (DMSO), 2 g/kg [D3] acetate bolus
SA37807004 BT-474 Xeno D3 Acetate 6 hrs Plasma DMSO 1089BT-474 Xenograft in an NSG mouse Tumor Vehicle (DMSO), 2 g/kg [D3] acetate bolus
SA37807153 Fasnall PK 1900 3 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807245 Fasnall PK 1908 6 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807346 Fasnall PK 1907 6 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807447 Fasnall PK 1906 6 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807548 Fasnall PK 1905 4 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807649 Fasnall PK 1904 4 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807750 Fasnall PK 1903 4 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807851 Fasnall PK 1902 3 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37807952 Fasnall PK 1901 3 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808059 Fasnall PK 1894 1 hrMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808154 Fasnall PK 1899 2 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808255 Fasnall PK 1898 2 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808356 Fasnall PK 1897 2 hrsMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808457 Fasnall PK 1896 1 hrMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808558 Fasnall PK 1895 1 hrMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808660 Fasnall PK 1892 15minMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808761 Fasnall PK 1891 15minMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808862 Fasnall PK 1892 15minMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37808963 Fasnall PK 1891 15minMurine plasma - Pharmacokinetics experiment Plasma 10 mg/kg Fasnall
SA37809039 Plasma Fasnall Std 2.5 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809143 Plasma Fasnall Std 50 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809242 Plasma Fasnall Std 25 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809341 Plasma Fasnall Std 10 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809440 Plasma Fasnall Std 5 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809534 Plasma Fasnall Std 20 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809638 Plasma Fasnall Std 1 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809737 Plasma Fasnall Std 500 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809815 Plasma Fasnall Std 0.2 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37809916 Plasma Fasnall Std 1 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810017 Plasma Fasnall Std 2 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810118 Plasma Fasnall Std 10 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810219 Plasma Fasnall Std 20 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810320 Plasma Fasnall Std 100 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810421 Plasma Fasnall Std 250 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810522 Plasma Fasnall Std 500 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810623 Plasma Fasnall Std 1 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810724 Plasma Fasnall Std 2.5 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810825 Plasma Fasnall Std 5 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37810926 Plasma Fasnall Std 10 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811027 Plasma Fasnall Std 25 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811128 Plasma Fasnall Std 50 ug/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811230 Plasma Fasnall Std 0.2 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811331 Plasma Fasnall Std 1 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811432 Plasma Fasnall Std 2 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811533 Plasma Fasnall Std 10 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811635 Plasma Fasnall Std 100 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811736 Plasma Fasnall Std 250 ng/mlMurine plasma spiked with Fasnall Plasma-based standard NA
SA37811829 BlankNA Blank NA
SA37811944 BlankNA Blank NA
SA37812064 BlankNA Blank NA
SA37812166 BlankNA Blank NA
SA37812201 BlankNA Blank NA
SA37812302 DMEM/F12 no addsNA QC NA
SA37812465 DMEM/F12 no addsNA QC NA
Showing results 1 to 66 of 66

Collection:

Collection ID:CO003554
Collection Summary:Plasma metabolomics samples were prepared from whole blood mixed with EDTA to 10 mM final concentration. To separate plasma, blood samples were centrifuged at 2,000 g and 4 °C for 10 min. Then, 50 µl of plasma were mixed with 450 µl of 100% methanol to reach 90% final methanol concentration. 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. Regardless of the type of metabolite extracts, 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.
Collection Protocol Filename:DM_metabolomics_samples.txt
Sample Type:Blood (plasma)
Collection Method:90% methanol extraction
Storage Conditions:-80℃
Collection Vials:1.5 ml plastic centrifuge tubes
Storage Vials:1.5 ml plastic centrifuge tubes

Treatment:

Treatment ID:TR003570
Treatment Summary:A single dose of 10 mg/kg (drug/body weight) Fasnall was injected intraperitoneally (~50 ul of solution in 1:1 DMSO:PBS), and plasma samples were collected. Blood was collected through a cardiac puncture. EDTA was used as an anticoagulant.
Treatment:Drug bolus
Treatment Compound:Fasnall
Treatment Route:Intraperitoneal injection
Treatment Dose:10 mg/kg
Treatment Dosevolume:~50 ul
Treatment Doseduration:Single dose, 6 h monitoring
Treatment Vehicle:1:1 DMSO:PBS
Animal Endp Euthanasia:CO2
Animal Endp Tissue Coll List:Blood

Sample Preparation:

Sampleprep ID:SP003568
Sampleprep Summary:Plasma metabolomics samples were prepared from whole blood mixed with EDTA to 10 mM final concentration. To separate plasma, blood samples were centrifuged at 2,000 g and 4 °C for 10 min. Then, 50 µl of plasma were mixed with 450 µl of 100% methanol to reach 90% final methanol concentration. 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. Regardless of the type of metabolite extracts, 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
Processing Storage Conditions:4℃
Extraction Method:90% methanol
Extract Enrichment:None
Extract Cleanup:None
Extract Storage:-80℃
Sample Resuspension:None
Sample Derivatization:None
Sample Spiking:None
Subcellular Location:Intracellular metabolites and medium metabolites

Combined analysis:

Analysis ID AN005641 AN005642
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Shimadzu 20AD Shimadzu 20AD
Column Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um) Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um)
MS Type ESI ESI
MS instrument type Triple quadrupole Triple quadrupole
MS instrument name ABI Sciex 5500 QTrap ABI Sciex 5500 QTrap
Ion Mode NEGATIVE POSITIVE
Units Counts per second (cps) Counts per second (cps)

Chromatography:

Chromatography ID:CH004283
Instrument Name:Shimadzu 20AD
Column Name:Merck SeQuant ZIC-HILIC (150 x 2.1mm,5um)
Column Pressure:900-3000 psi
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
Injection Temperature:4
Sample Injection:1-5 ul
Solvent A:100% Water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate
Solvent B:100% Acetonitrile
Chromatography Type:HILIC

MS:

MS ID:MS005365
Analysis ID:AN005641
Instrument Name:ABI Sciex 5500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Data were analyzed with SCIEX Multiquant 3.0.3.
Ion Mode:NEGATIVE
Capillary Temperature:500 °C
Capillary Voltage:-4500
Dry Gas Flow:70
Dry Gas Temp:500 °C
Ion Source Temperature:500 °C
Ion Spray Voltage:-4500
Mass Accuracy:UNIT
Source Temperature:500 °C
Spray Voltage:-4500
Desolvation Gas Flow:70
Desolvation Temperature:500 °C
  
MS ID:MS005366
Analysis ID:AN005642
Instrument Name:ABI Sciex 5500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:Data were analyzed with SCIEX Multiquant 3.0.3.
Ion Mode:POSITIVE
Capillary Temperature:500 °C
Capillary Voltage:4500
Dry Gas Flow:70
Dry Gas Temp:500 °C
Ion Source Temperature:500 °C
Ion Spray Voltage:4500
Mass Accuracy:UNIT
Source Temperature:500 °C
Spray Voltage:4500
Desolvation Gas Flow:70
Desolvation Temperature:500 °C
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