{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003185","ANALYSIS_ID":"AN005232","VERSION":"1","CREATED_ON":"May 3, 2024, 10:13 am"},

"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 - plasma lipidomics","STUDY_SUMMARY":"Lipidomics profiling of immunocompetent B16F10 model of melanoma to examine lipid levels in mouse plasma 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":"DS2-039-021",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-021"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-022",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-022"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-023",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-023"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-024",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-024"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-025",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-025"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-026",
"Factors":{"factor":"vehicle + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-026"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-027",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-027"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-028",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-028"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-029",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-029"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-030",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-030"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-031",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-031"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-032",
"Factors":{"factor":"SCD inhibitor + low oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-032"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-033",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-033"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-034",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-034"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-035",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-035"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-036",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-036"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-037",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-037"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-038",
"Factors":{"factor":"Vehicle + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-038"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-039",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-039"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-040",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-040"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-041",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-041"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-042",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-042"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-043",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-043"}
},
{
"Subject ID":"-",
"Sample ID":"DS2-039-044",
"Factors":{"factor":"SCD inhibitor + high oleic acid diet","Sample source":"plasma"},
"Additional sample data":{"RAW_FILE_NAME(raw file name)":"DS2-039-044"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"After two weeks of treatment, retroorbital whole blood samples were collected following Isoflurane sedation. The samples were obtained using heparinized capillary tubes (Catalog Number: 22-362566). Subsequently, these samples were placed into tubes containing K2E (Catalog Number: 365974), with a bulb used to drain the capillary tube (Catalog Number: 22-362566). The tubes were centrifuged for 15 minutes at 2,000 x g (4°C). The resulting plasma (supernatant) was collected and aliquoted into 1.5ml Eppendorf tubes and stored at -80°C.","SAMPLE_TYPE":"Blood (plasma)"},

"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. Treatment began three days after tumor cell implantation (bilateral tumors in n=4 per group). Mice were euthanized17 days after tumor cell injection/14 days after the start of treatment."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"For plasma analysis, 20 µL of plasma was aliquoted into 1.5 mL Eppendorf tubes followed by 480 µL cold MeOH. Samples were then vortexed for 30 minutes, 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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}

}