Summary of Study ST002865
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 PR001786. The data can be accessed directly via it's Project DOI: 10.21228/M8VB1G 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 | ST002865 |
| Study Title | Metabolic profiling, glucose tracing and glutamine tracing in 16D prostate cancer cells treated with vehicle, AR inhibitor Enzalutamide, AR inhibitor Apalutamide, or AR degrader/PROTAC ARCC-4 |
| Study Summary | Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer. In the MS data, M0, M1, M2, M3,... represent isotopologues of each metabolite. |
| Institute | University of California, Los Angeles |
| Department | Molecular, Cell and Developmental Biology |
| Laboratory | Andrew Goldstein |
| Last Name | Goldstein |
| First Name | Andrew |
| Address | 610 Charles E Young Dr East, Goldstein Lab 3141 Terasaki Life Sci Bld, Los Angeles, CA, 90095, USA |
| AGoldstein@mednet.ucla.edu | |
| Phone | 3102061402 |
| Submit Date | 2023-09-12 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2023-09-19 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001786 |
| Project DOI: | doi: 10.21228/M8VB1G |
| Project Title: | MYC is a regulator of androgen receptor inhibition-induced metabolic requirements in prostate cancer |
| Project Summary: | Advanced prostate cancers are treated with therapies targeting the androgen receptor (AR) signaling pathway. While many tumors initially respond to AR inhibition, nearly all develop resistance. It is critical to understand how prostate tumor cells respond to AR inhibition in order to exploit therapy-induced phenotypes prior to the outgrowth of treatment-resistant disease. Here, we comprehensively characterize the effect of AR blockade on prostate cancer metabolism using transcriptomics, metabolomics and bioenergetics approaches. The metabolic response to AR inhibition is defined by reduced glycolysis, robust elongation of mitochondria, and increased reliance on mitochondrial oxidative metabolism. We establish DRP1 activity and MYC signaling as mediators of AR blockade-induced metabolic phenotypes. Rescuing DRP1 phosphorylation after AR inhibition restores mitochondrial fission, while rescuing MYC restores glycolytic activity and prevents sensitivity to complex I inhibition. Our study provides new insight into the regulation of treatment-induced metabolic phenotypes and vulnerabilities in prostate cancer. |
| Institute: | University of California, Los Angeles |
| Department: | Biological Chemistry |
| Laboratory: | Heather Christofk |
| Last Name: | Matulionis |
| First Name: | Nedas |
| Address: | 615 Charles E Young Dr S, BSRB 354-05 |
| Email: | nmatulionis@mednet.ucla.edu |
| Phone: | 3102060163 |
Subject:
| Subject ID: | SU002977 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment |
|---|---|---|
| SA309483 | NN.009 | Apalutamide_C13Glc |
| SA309484 | NN.008 | Apalutamide_C13Glc |
| SA309485 | NN.007 | Apalutamide_C13Glc |
| SA309486 | NN.019 | Apalutamide_C13Gln |
| SA309487 | NN.021 | Apalutamide_C13Gln |
| SA309488 | NN.020 | Apalutamide_C13Gln |
| SA309477 | NN.012 | ARCC4_C13Glc |
| SA309478 | NN.010 | ARCC4_C13Glc |
| SA309479 | NN.011 | ARCC4_C13Glc |
| SA309480 | NN.022 | ARCC4_C13Gln |
| SA309481 | NN.024 | ARCC4_C13Gln |
| SA309482 | NN.023 | ARCC4_C13Gln |
| SA309489 | NN.005 | Enzalutamide_C13Glc |
| SA309490 | NN.004 | Enzalutamide_C13Glc |
| SA309491 | NN.006 | Enzalutamide_C13Glc |
| SA309492 | NN.018 | Enzalutamide_C13Gln |
| SA309493 | NN.017 | Enzalutamide_C13Gln |
| SA309494 | NN.016 | Enzalutamide_C13Gln |
| SA309495 | QC.blank1 | NA |
| SA309496 | QC.blank2 | NA |
| SA309497 | QC.blank3 | NA |
| SA309498 | NN.003 | Vehicle_C13Glc |
| SA309499 | NN.001 | Vehicle_C13Glc |
| SA309500 | NN.002 | Vehicle_C13Glc |
| SA309501 | NN.014 | Vehicle_C13Gln |
| SA309502 | NN.013 | Vehicle_C13Gln |
| SA309503 | NN.015 | Vehicle_C13Gln |
| Showing results 1 to 27 of 27 |
Collection:
| Collection ID: | CO002970 |
| Collection Summary: | Media was aspirated and cells were washed with cold 150mM ammonium acetate pH 7.3. Metabolite extractions were performed by adding 500μl of cold 80% methanol containing 2nM Norvaline (Sigma) as an internal standard per well. Cells were removed using a cell scraper before transferring cell suspensions to 1.5ml Eppendorf tubes. Samples were vortexed for 30 seconds and spun at 4°C for 5 minutes at maximum speed to pellet the insoluble fraction before 420 µl of the soluble fraction was transferred to ABC vials (Thermo Fisher Scientific). 80% MeOH was evaporated from the ABC vials using the EZ-2Elite evaporator (Genevac) and samples were stored at -80°C until analysis. |
| Sample Type: | Prostate cancer cells |
Treatment:
| Treatment ID: | TR002986 |
| Treatment Summary: | Tissue culture plates were coated with 0.01% (v/v) Poly-L-Lysine (Sigma, P4832) diluted 1/20 in distilled water and washed with PBS to enhance cell attachment. 16D cells were cultured in RPMI base media (Gibco) + 10% FBS (v/v) + 100 units/mL penicillin, and 100μg/mL streptomycin. Treatment was performed by adding 10μM Enzalutamide, 10uM Apalutamide, or 0.5uM ARCC4 every 48 hours. |
| Treatment Compound: | Enzalutamide |
| Treatment Vehicle: | DMSO |
Sample Preparation:
| Sampleprep ID: | SP002983 |
| Sampleprep Summary: | For metabolite extraction, media was aspirated and cells were washed with cold 150mM ammonium acetate pH 7.3. Metabolite extractions were performed by adding 500μl of cold 80% methanol containing 2nM Norvaline (Sigma) as an internal standard per well. Cells were removed using a cell scraper before transferring cell suspensions to 1.5ml Eppendorf tubes. Samples were vortexed for 30 seconds and spun at 4°C for 5 minutes at maximum speed to pellet the insoluble fraction before 420 µl of the soluble fraction was transferred to ABC vials (Thermo Fisher Scientific). 80% MeOH was evaporated from the ABC vials using the EZ-2Elite evaporator (Genevac) and samples were stored at -80°C until analysis. |
Combined analysis:
| Analysis ID | AN004697 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish |
| Column | Phenomenex Luna NH2 (150 x 2mm,3um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive Orbitrap |
| Ion Mode | UNSPECIFIED |
| Units | Abundance |
Chromatography:
| Chromatography ID: | CH003537 |
| Chromatography Summary: | Dried metabolites were resuspended in 50% ACN:water and 1/10th was loaded onto a Luna 3um NH2 100A (150 × 2.0 mm) column (Phenomenex). The chromatographic separation was performed on a Vanquish Flex (Thermo Fisher Scientific) with mobile phases A (5 mM NH4AcO pH 9.9) and B (ACN) and a flow rate of 200 μl/minute. A linear gradient from 15% A to 95% A over 18 minutes was followed by 9 minutes isocratic flow at 95% A and reequilibration to 15% A. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Phenomenex Luna NH2 (150 x 2mm,3um) |
| Column Temperature: | 35 |
| Flow Gradient: | Linear gradient was as follows: 15% A to 95% A over 18 minutes was followed by 9 minutes isocratic flow at 95% A and reequilibration to 15% A |
| Flow Rate: | 200 ul/minute |
| Solvent A: | 5 mM NH4AcO pH 9.9 |
| Solvent B: | ACN |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004444 |
| Analysis ID: | AN004697 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Metabolites were detection with a Thermo Fisher Scientific Q Exactive mass spectrometer run with polarity switching (+3.5 kV/− 3.5 kV) in full scan mode with an m/z range of 70-975 and 70.000 resolution. TraceFinder 4.1 (Thermo Fisher Scientific) was used to quantify the targeted metabolites by area under the curve using expected retention time and accurate mass measurements (< 5 ppm). |
| Ion Mode: | UNSPECIFIED |
