{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003184","ANALYSIS_ID":"AN005229","VERSION":"1","CREATED_ON":"May 2, 2024, 6:17 pm"},

"PROJECT":{"PROJECT_TITLE":"A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis","PROJECT_SUMMARY":"The genetic landscape of cancer cells can lead to specific metabolic dependencies for tumor growth. Dietary interventions represent an attractive strategy to restrict the availability of key nutrients to tumors. In this study, we identified that growth of a subset of melanoma was severely restricted by a rationally designed combination therapy of a stearoyl-CoA desaturase (SCD) inhibitor with an isocaloric low-oleic acid diet. Despite its importance in oncogenesis, SCD underwent monoallelic co-deletion along with PTEN on chromosome 10q in about 47.5% of melanoma, and the other SCD allele was methylated, resulting in very low SCD expression. While this SCD deficient subset was refractory to SCD inhibitors, the subset of PTEN wildtype melanoma that retained SCD was sensitive. As dietary oleic acid could potentially blunt the effect of SCD inhibitors, a low-oleic acid custom diet was combined with SCD inhibitor. The combination reduced monounsaturated fatty acids and increased saturated fatty acids, inducing robust apoptosis and growth suppression and inhibiting lung metastasis with minimal toxicity in preclinical mouse models of PTEN wildtype melanoma. When combined with anti-PD1 immunotherapy, the SCD inhibitor improved T cell functionality and further constrained melanoma growth in mice. Collectively, these results suggest that optimizing SCD inhibitors with diets low in oleic acid may offer a viable and efficacious therapeutic approach for improving melanoma treatment.","INSTITUTE":"University of Colorado Anschutz Medical Campus","LABORATORY":"Lab of Angelo D''Alessandro in collaboration with lab of Biplab Dasgupta","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"STUDY":{"STUDY_TITLE":"A multimodal drug-diet-immunotherapy combination restrains melanoma progression and metastasis - tumor lipidomics","STUDY_SUMMARY":"Lipidomics profiling of immunocompetent B16F10 model of melanoma to examine lipid levels in tumors following reduced oleic acid content in diet combined with the stearyl CoA desaturase inhibitor CAY10566.","INSTITUTE":"University of Colorado Anschutz Medical Campus","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"SUBJECT":{"SUBJECT_TYPE":"Mammal","SUBJECT_SPECIES":"Mus musculus","GENDER":"Not applicable"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"DS2-039-001",
"Factors":{"factor":"Vehicle+ low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-001"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-002",
"Factors":{"factor":"Vehicle+ low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-002"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-003",
"Factors":{"factor":"Vehicle+ low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-003"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-004",
"Factors":{"factor":"Vehicle+ low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-004"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-005",
"Factors":{"factor":"Vehicle+ low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-005"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-006",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-006"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-007",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-007"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-008",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-008"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-009",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-009"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-010",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-010"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-011",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-011"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-012",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-012"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-013",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-013"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-014",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-014"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-015",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-015"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-016",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-016"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-017",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-017"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-018",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-018"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-019",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-019"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-020",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"tumor"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-020"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"For immune checkpoint inhibitor (ICI) studies, 1x10^6 B16F10 cells were mixed with matrigel 1:1 and implanted subcutaneously in each flank into C57BL/6 mice. Mice were randomized into groups with combinations of sunflower or coconut oil diet, vehicle or CAY10566, and IgG or anti-PD1 treatments. CAY10566 was prepared and administered as previously described. IgG and anti-PD1 were diluted in PBS to a concentration of 1ug/uL. 100uL of IgG or anti-PD1 were administered intraperitoneally twice weekly, every 3-4 days. Mice were euthanized and tumors were harvested 17 days after tumor implantation/14 days after the start of treatment. Tumor tissues were processed for flow cytometry analysis. For lung metastasis studies, ZsGreen expressing B16F10 cells were established by transducing them with concentrated ZsGreen virus produced by the Viral Vector Core at CCHMC. Cells were then FACS sorted to obtain a pure population of B16F10-ZsGreen cells prior to injection. 2x10^5 B16F10-ZsGreen cells were injected via tail vein into C57BL/6 mice by trained veterinary staff of CCHMC. Mice were randomized into groups with combinations of sunflower or coconut oil diet, and vehicle or CAY10566. CAY10566 was prepared and administered as previously described. Mice were euthanized and lungs were harvested 17 days after tumor cell injection/14 days after the start of treatment. B16F10-ZsGreen metastatic tumor nodes were detected by fluorescence microscopy, and quantification was performed using Image J.","SAMPLE_TYPE":"Tumor cells"},

"TREATMENT":{"TREATMENT_SUMMARY":"All animal procedures were carried out in accordance with the IACUC-approved protocol of Cincinnati Children’s Hospital Medical Center. Animals were monitored daily by Veterinary Services. For subcutaneous tumor implantation, 1x10^6 B16F10 cells were mixed with matrigel 1:1 and implanted subcutaneously into NOD-SCID IL2Rgnull or C57BL/6 mice, respectively. CAY10566 was suspended in 0.5% Methyl Cellulose in water with 0.2% Tween 80 at 9 parts per 1 part DMSO stock. CAY10566 was administered at 30mg/kg via oral gavage, twice daily, with a drug holiday during the weekends."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For tumor analysis, tissue was weighed and placed into 1.5mL Eppendorf Safe-Lock tubes that were pre-filled with 1.0mm glass grinder beads (Biospec Products Inc). Cold methanol was then added to each tube so that all final tissue concentrations were 20 mg/mL. Samples were then homogenized via a “Next Advance” Bullet Blender for 5 minutes and subsequently vortexed for 30 minutes. Following vortex, insoluble material was pelleted by centrifugation (18000g, 10 min) and supernatants were isolated for analysis by UHPLC-MS.","PROCESSING_STORAGE_CONDITIONS":"4℃","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Positive C18 global lipidomics","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)","SOLVENT_A":"75% water/25% acetonitrile; 5 mM ammonium acetate","SOLVENT_B":"90% isopropanol/10% acetonitrile; 5 mM ammonium acetate","FLOW_GRADIENT":"0-3 min 30-100% B at 0.3 mL/min, 3-4.2 min hold at 100% B at 0.3 mL/min, 4.2-4.3 min 100-30% B at 0.4 mL/min, 4.3-4.9 min hold at 30%B and 0.4 mL/min, 4.9-5 min hold at 30%B while lowering flow rate to 0.3 mL/min","FLOW_RATE":"0.3-0.4 ml/min","COLUMN_TEMPERATURE":"45","SAMPLE_INJECTION":"10 uL"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control."},

"MS_METABOLITE_DATA":{
"Units":"peak area",

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}