Summary of Study ST002863

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	ST002863
Study Title	Metabolic profiling and glucose tracing in naive and Enzalutamide-treated 16D prostate cancer cells
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
Email	AGoldstein@mednet.ucla.edu
Phone	3102061402
Submit Date	2023-09-11
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:	SU002975
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
SA309436	JG-004	Enzalutamide 24 hours
SA309437	JG-006	Enzalutamide 24 hours
SA309438	JG-005	Enzalutamide 24 hours
SA309439	JG-009	Enzalutamide 48 hours
SA309440	JG-007	Enzalutamide 48 hours
SA309441	JG-008	Enzalutamide 48 hours
SA309442	JG-012	Enzalutamide long-term
SA309443	JG-010	Enzalutamide long-term
SA309444	JG-011	Enzalutamide long-term
SA309445	QC-blank2	NA
SA309446	QC-blank1	NA
SA309447	JG-002	vehicle (DMSO)
SA309448	JG-003	vehicle (DMSO)
SA309449	JG-001	vehicle (DMSO)

Showing results 1 to 14 of 14

Showing results 1 to 14 of 14

Collection:

Collection ID:	CO002968
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:	TR002984
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 and LNCaP cells were cultured in RPMI base media (Gibco) + 10% FBS (v/v) + 100 units/mL penicillin, and 100μg/mL streptomycin. Enzalutamide treatment was performed by adding 10μM Enzalutamide (Selleck Chemicals, S1250) every 48 hours.
Treatment Compound:	Enzalutamide
Treatment Vehicle:	DMSO

Sample Preparation:

Sampleprep ID:	SP002981
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.

Sampleprep ID:

SP002981

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	AN004695
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:	CH003535
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:	MS004442
Analysis ID:	AN004695
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