Summary of Study ST003436
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 | ST003436 |
Study Title | Pharmacokinetics of Fasnall in NSG mice |
Study Summary | Fasnall concentrations in plasma, heart, liver, and brain of NSG mice were monitored after 10 mg/kg intraperitoneal bolus. |
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 | +12154956903 |
Submit Date | 2024-08-21 |
Num Groups | 27 |
Total Subjects | 82 |
Publications | Submission Pending |
Raw Data Available | Yes |
Raw Data File Type(s) | mzML, wiff |
Analysis Type Detail | LC-MS |
Release Date | 2024-09-12 |
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: | SU003563 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Genotype Strain: | NOD Scid Gamma (NSG) |
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 |
Species Group: | Mammals |
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 | Time post-injection |
---|---|---|---|---|---|
SA378174 | 72 F12 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378175 | 60 F10 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378176 | 61 F09 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378177 | 62 F08 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378178 | 63 F07 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378179 | 64 F06 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378180 | 66 F04 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378181 | 67 F03 Blue 25 uM | FBS-based standard | FBS-based standard | NA | NA |
SA378182 | 68 F02 Blue 50 uM | FBS-based standard | FBS-based standard | NA | NA |
SA378183 | 69 F01 Blue 100 uM Fasnall in dFBS | FBS-based standard | FBS-based standard | NA | NA |
SA378184 | 73 F11 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378185 | 58 F12 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378186 | 74 F10 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378187 | 75 F09 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378188 | 76 F08 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378189 | 77 F07 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378190 | 78 F06 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378191 | 79 F05 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378192 | 80 F04 Red | FBS-based standard | FBS-based standard | NA | NA |
SA378193 | 81 F03 Red 25 uM | FBS-based standard | FBS-based standard | NA | NA |
SA378194 | 82 F02 Red 50 uM | FBS-based standard | FBS-based standard | NA | NA |
SA378195 | 83 F01 Red 100 uM Fasnall in dFBS | FBS-based standard | FBS-based standard | NA | NA |
SA378196 | 59 F11 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378197 | 65 F05 Blue | FBS-based standard | FBS-based standard | NA | NA |
SA378201 | 126 Fasnall PK 1891 15 min Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378202 | 125 Fasnall PK 1892 15 min Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378203 | 124 Fasnall PK 1893 15 min Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378198 | 122 Fasnall PK 1895 1 hr Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378199 | 123 Fasnall PK 1894 1 hr Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378200 | 121 Fasnall PK 1896 1 hr Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378204 | 118 Fasnall PK 1899 2 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378205 | 120 Fasnall PK 1897 2 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378206 | 119 Fasnall PK 1898 2 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378207 | 115 Fasnall PK 1902 3 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378208 | 116 Fasnall PK 1901 3 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378209 | 117 Fasnall PK 1900 3 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378210 | 112 Fasnall PK 1905 4 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378211 | 113 Fasnall PK 1904 4 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378212 | 114 Fasnall PK 1903 4 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378213 | 109 Fasnall PK 1908 6 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378214 | 110 Fasnall PK 1907 6 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378215 | 111 Fasnall PK 1906 6 hrs Brain | Murine brain | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378216 | 108 Fasnall PK Control Brain | Murine brain | Tissue metabolite extract | Control | NA |
SA378217 | 107 Fasnall PK Control Brain | Murine brain | Tissue metabolite extract | Control | NA |
SA378218 | 106 Fasnall PK Control Brain | Murine brain | Tissue metabolite extract | Control | NA |
SA378222 | 57 Fasnall PK 1891 15 min Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378223 | 55 Fasnall PK 1893 15 min Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378224 | 56 Fasnall PK 1892 15 min Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378219 | 53 Fasnall PK 1895 1 hr Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378220 | 52 Fasnall PK 1896 1 hr Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378221 | 54 Fasnall PK 1894 1 hr Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378225 | 49 Fasnall PK 1899 2 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378226 | 50 Fasnall PK 1898 2 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378227 | 51 Fasnall PK 1897 2 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378228 | 47 Fasnall PK 1901 3 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378229 | 48 Fasnall PK 1900 3 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378230 | 46 Fasnall PK 1902 3 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378231 | 44 Fasnall PK 1904 4 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378232 | 43 Fasnall PK 1905 4 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378233 | 45 Fasnall PK 1903 4 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378234 | 42 Fasnall PK 1906 6 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378235 | 41 Fasnall PK 1907 6 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378236 | 40 Fasnall PK 1908 6 hrs Heart | Murine heart | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378237 | 39 Fasnall PK Control Heart | Murine heart | Tissue metabolite extract | Control | NA |
SA378238 | 38 Fasnall PK Control Heart | Murine heart | Tissue metabolite extract | Control | NA |
SA378239 | 37 Fasnall PK Control Heart | Murine heart | Tissue metabolite extract | Control | NA |
SA378243 | 104 Fasnall PK 1892 15 min Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378244 | 103 Fasnall PK 1893 15 min Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378245 | 105 Fasnall PK 1891 15 min Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 15 min |
SA378240 | 101 Fasnall PK 1895 1 hr Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378241 | 100 Fasnall PK 1896 1 hr Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378242 | 102 Fasnall PK 1894 1 hr Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 1 h |
SA378246 | 99 Fasnall PK 1899 2 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378247 | 98 Fasnall PK 1897 2 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378248 | 97 Fasnall PK 1899 2 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 2 h |
SA378249 | 95 Fasnall PK 1901 3 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378250 | 96 Fasnall PK 1900 3 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378251 | 94 Fasnall PK 1902 3 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 3 h |
SA378252 | 91 Fasnall PK 1905 4 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378253 | 93 Fasnall PK 1903 4 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378254 | 92 Fasnall PK 1904 4 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 4 h |
SA378255 | 88 Fasnall PK 1908 6 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378256 | 89 Fasnall PK 1907 6 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378257 | 90 Fasnall PK 1906 6 hrs Liver | Murine liver | Tissue metabolite extract | 10 mg/kg Fasnall | 6 h |
SA378258 | 85 Fasnall PK Control Liver | Murine liver | Tissue metabolite extract | Control | NA |
SA378259 | 86 Fasnall PK Control Liver | Murine liver | Tissue metabolite extract | Control | NA |
SA378260 | 87 Fasnall PK Control Liver | Murine liver | Tissue metabolite extract | Control | NA |
SA378261 | 16 Plasma Fasnall Std 50 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378262 | 15 Plasma Fasnall Std 25 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378263 | 14 Plasma Fasnall Std 10 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378264 | 03 Plasma Fasnall Std 0.2 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378265 | 04 Plasma Fasnall Std 1 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378266 | 05 Plasma Fasnall Std 2 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378267 | 06 Plasma Fasnall Std 10 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378268 | 07 Plasma Fasnall Std 20 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378269 | 09 Plasma Fasnall Std 250 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378270 | 10 Plasma Fasnall Std 500 ng/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378271 | 11 Plasma Fasnall Std 1 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378272 | 12 Plasma Fasnall Std 2.5 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
SA378273 | 13 Plasma Fasnall Std 5 ug/ml | Murine plasma | Plasma-based standard | NA | NA |
Collection:
Collection ID: | CO003556 |
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: | Brain, Heart, Liver, Blood(plasma) |
Collection Method: | 80% methanol extraction for organs, 90% for plasma |
Storage Conditions: | -80℃ |
Collection Vials: | 1.5 ml plastic centrifuge tubes |
Storage Vials: | 1.5 ml plastic centrifuge tubes |
Treatment:
Treatment ID: | TR003572 |
Treatment Summary: | Cells were grown in RPMI-1640 supplemented with 10% dialyzed FBS and treated with various concentrations of dimethylmalonate (DMM) for 24 h. |
Treatment Compound: | Fasnall (1-benzyl-N-{5,6-dimethylthieno[2,3-d]pyrimidin-4-yl}pyrrolidin-3-amine, Enamine EN300-14327544) |
Treatment Vehicle: | 1:1 DMSO:PBS |
Sample Preparation:
Sampleprep ID: | SP003570 |
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: | 80% methanol for organs, 90% for plasma |
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 | AN005645 | AN005646 |
---|---|---|
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: | CH004285 |
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: | MS005369 |
Analysis ID: | AN005645 |
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: | MS005370 |
Analysis ID: | AN005646 |
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 |