{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST002863","ANALYSIS_ID":"AN004695","VERSION":"1","CREATED_ON":"September 14, 2023, 9:59 am"},

"PROJECT":{"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":"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"},

"STUDY":{"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.","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"},

"SUBJECT":{"SUBJECT_TYPE":"Cultured cells","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"JG-001",
"Factors":{"Treatment":"vehicle (DMSO)"},
"Additional sample data":{"RAW_FILE_NAME":"JG-001.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-002",
"Factors":{"Treatment":"vehicle (DMSO)"},
"Additional sample data":{"RAW_FILE_NAME":"JG-002.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-003",
"Factors":{"Treatment":"vehicle (DMSO)"},
"Additional sample data":{"RAW_FILE_NAME":"JG-003.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-004",
"Factors":{"Treatment":"Enzalutamide 24 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-004.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-005",
"Factors":{"Treatment":"Enzalutamide 24 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-005.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-006",
"Factors":{"Treatment":"Enzalutamide 24 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-006.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-007",
"Factors":{"Treatment":"Enzalutamide 48 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-007.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-008",
"Factors":{"Treatment":"Enzalutamide 48 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-008.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-009",
"Factors":{"Treatment":"Enzalutamide 48 hours"},
"Additional sample data":{"RAW_FILE_NAME":"JG-009.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-010",
"Factors":{"Treatment":"Enzalutamide long-term"},
"Additional sample data":{"RAW_FILE_NAME":"JG-010.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-011",
"Factors":{"Treatment":"Enzalutamide long-term"},
"Additional sample data":{"RAW_FILE_NAME":"JG-011.raw"}
},
{
"Subject ID":"-",
"Sample ID":"JG-012",
"Factors":{"Treatment":"Enzalutamide long-term"},
"Additional sample data":{"RAW_FILE_NAME":"JG-012.raw"}
},
{
"Subject ID":"-",
"Sample ID":"QC-blank1",
"Factors":{"Treatment":"NA"},
"Additional sample data":{"RAW_FILE_NAME":"QC-blank1.raw"}
},
{
"Subject ID":"-",
"Sample ID":"QC-blank2",
"Factors":{"Treatment":"NA"},
"Additional sample data":{"RAW_FILE_NAME":"QC-blank2.raw"}
}
],
"COLLECTION":{"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_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"},

"SAMPLEPREP":{"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."},

"CHROMATOGRAPHY":{"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.","CHROMATOGRAPHY_TYPE":"HILIC","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Luna NH2 (150 x 2mm,3um)","SOLVENT_A":"5 mM NH4AcO pH 9.9","SOLVENT_B":"ACN","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","COLUMN_TEMPERATURE":"35"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"UNSPECIFIED","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)."},

"MS_METABOLITE_DATA":{
"Units":"Abundance",

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}

}