{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001903","ANALYSIS_ID":"AN003815","VERSION":"1","CREATED_ON":"November 8, 2022, 11:35 am"},
"PROJECT":{"PROJECT_TITLE":"Effect of ketogenic diet on the plasma and tumor metabolome of melanoma-bearing mice","PROJECT_TYPE":"Metabolomics analysis","PROJECT_SUMMARY":"Growing evidence supports the use of low-carbohydrate/high-fat ketogenic diets (KDs) together with standard therapies to improve cancer treatment outcomes. However, conflicting data exist regarding the efficacy of KDs in melanoma, the deadliest skin cancer. Here, we show that two different KD formulations effectively reduced tumor growth in immunocompromised mice bearing genetically and metabolically heterogeneous human melanoma xenografts. Furthermore, the KDs exerted a metastasis-reducing effect in an immunocompetent syngeneic melanoma mouse model. Ketone bodies did not directly influence melanoma cell proliferation; therefore, we performed an in-depth metabolomics analysis using the MxP® Quant 500 kit combined with a acylcarnitine assay (biocrates life sciences ag)to elucidate potential antitumor mechanisms in vivo. Targeted analysis of plasma and tumor metabolomes revealed distinct changes in amino acid metabolism induced by the KDs. Moreover, the KDs increased sphingomyelin synthesis and hydroxylation of certain lipids. Thus, KDs simultaneously affect multiple metabolic pathways to create an unfavorable environment for melanoma cell proliferation, supporting their potential as a complementary nutritional approach to melanoma therapy.","INSTITUTE":"University Hospital of the Paracelsus Medical University Salzburg","LAST_NAME":"Weber","FIRST_NAME":"Daniela","ADDRESS":"Müllner Hauptstraße 48, Salzburg, Salzburg, 5020, Austria","EMAIL":"d.weber@salk.at","PHONE":"0043 57255 26274"},
"STUDY":{"STUDY_TITLE":"Effect of ketogenic diet on the plasma and tumor metabolome of melanoma-bearing mice","STUDY_SUMMARY":"Growing evidence supports the use of low-carbohydrate/high-fat ketogenic diets (KDs) together with standard therapies to improve cancer treatment outcomes. However, conflicting data exist regarding the efficacy of KDs in melanoma, the deadliest skin cancer. Here, we show that two different KD formulations effectively reduced tumor growth in immunocompromised mice bearing genetically and metabolically heterogeneous human melanoma xenografts. Furthermore, the KDs exerted a metastasis-reducing effect in an immunocompetent syngeneic melanoma mouse model. Ketone bodies did not directly influence melanoma cell proliferation; therefore, we performed an in-depth metabolomics analysis using the MxP® Quant 500 kit combined with a acylcarnitine assay (biocrates life sciences ag)to elucidate potential anti-tumor mechanisms in vivo. Targeted analysis of plasma and tumor metabolomes revealed distinct changes in amino acid metabolism induced by the KDs. Moreover, the KDs increased sphingomyelin synthesis and hydroxylation of certain lipids. Thus, KDs simultaneously affect multiple metabolic pathways to create an unfavorable environment for melanoma cell proliferation, supporting their potential as a complementary nutritional approach to melanoma therapy.","INSTITUTE":"University Hospital of the Paracelsus Medical University Salzburg","LAST_NAME":"Weber","FIRST_NAME":"Daniela","ADDRESS":"Müllner Hauptstraße 48, 5020 Salzburg, Austria","EMAIL":"d.weber@salk.at","STUDY_TYPE":"Metabolomics analysis","PHONE":"0043 57255 26274"},
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
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{
"Subject ID":"-",
"Sample ID":"tumor # 309",
"Factors":{"Sample description":"WM3000 LCT-MCT","Tissue":"tumor"},
"Additional sample data":{"RAW_FILE_NAME":"1020503613"}
},
{
"Subject ID":"-",
"Sample ID":"tumor # 310",
"Factors":{"Sample description":"WM3000 LCT-MCT","Tissue":"tumor"},
"Additional sample data":{"RAW_FILE_NAME":"1020503628"}
},
{
"Subject ID":"-",
"Sample ID":"tumor # 311",
"Factors":{"Sample description":"WM3000 LCT-MCT","Tissue":"tumor"},
"Additional sample data":{"RAW_FILE_NAME":"1020503632"}
},
{
"Subject ID":"-",
"Sample ID":"tumor # 312",
"Factors":{"Sample description":"WM3000 LCT-MCT","Tissue":"tumor"},
"Additional sample data":{"RAW_FILE_NAME":"1020503647"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Human melanoma xenografts were established in mice using A375 and WM47 (BRAF mutant), WM3311 (BRAF/NRAS wild-type) and WM3000 (NRAS mutant) cells. Blood was taken by cardiac puncture, collected in heparin-coated tubes and centrifuged for 3 min at 2000 g. Plasma samples were stored at -80 °C until analysis. Tumors were harvested and snap frozen in liquid nitrogen and stored at -80°C for metabolomics.","SAMPLE_TYPE":"Blood (plasma) and tumor tissue"},
"TREATMENT":{"TREATMENT_SUMMARY":"Human melanoma xenografts were established in mice using A375 and WM47 (BRAF mutant), WM3311 (BRAF/NRAS wild-type) and WM3000 (NRAS mutant) cells. Once the tumor volume reached 100 mm3, mice were equally assigned to control diet (CTRL) and two ketogenic diet (KDs) differing in their fat composition (LCT: long-chain triglyceride-based KD and LCT-MCT: long-chain triglyceride-based KD supplemented with C8 and C10 medium-chain triglycerides, ratio of fat to the sum of carbohydrate and protein of 8:1). Both KDs were administered ad libitum as monotherapy."},
"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"To capture a broad spectrum of metabolites, we used the MxP® Quant 500 kit combined with the in-house acylcarnitine (AC) assay (biocrates life sciences ag). Samples were prepared the same way for both assays. Therefore, 50-80 mg of tumor tissues were transferred to 0.5 ml Precellys® vials followed by the addition of 85:15 ethanol:0.01 M phosphate as a lysis buffer with a volume [µl] of 3 times the tissue weight [mg]. Tissue homogenization was carried out at 4 °C and 5800 rpm using a Precellys® 24 homogenizer coupled to a Cryolys® cooling unit. The homogenates were centrifuged at 10,000 g for 2 min at 2-4 °C. Supernatants were collected and stored at -80 °C until analysis. Plasma samples as well as tissue homogenate supernatant were thawed on ice, then vortexed before use."},
"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Waters Acquity","COLUMN_NAME":"MxP® Quant 500 Kit Column System"},
"ANALYSIS":{"ANALYSIS_TYPE":"MS"},
"MS":{"INSTRUMENT_NAME":"Waters TQ-S MS","INSTRUMENT_TYPE":"Triple quadrupole","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"MS acquisition data were obtained using MRM transition (Q1 and Q3 pair) in positive ion mode. Raw data of the MxP® Quant 500 assay were exported and quantified using MetIDQTM software (Biocrates Life Sciences). Downstream analysis was performed using MetaboAnalyst 5.0."},
"MS_METABOLITE_DATA":{
"Units":"uM",
"Data":[{"Metabolite":"1-Methylhistidine","plasma # 14":"2.936","plasma # 15":"3.754","plasma # 16":"4.667","plasma # 17":"4.316","plasma # 18":"2.795","plasma # 19":"2.491","plasma # 21":"3.287","plasma # 22":"3.064","plasma # 227":"5.499","plasma # 228":"3.206","plasma # 229":"4.504","plasma # 23":"5.942","plasma # 230":"4.007","plasma # 231":"5.057","plasma # 232":"2.819","plasma # 233":"5.513","plasma # 234":"4.449","plasma # 235":"4.988","plasma # 236":"2.902","plasma # 237":"3.33","plasma # 238":"2.888","plasma # 239":"2.542","plasma # 24":"4.012","plasma # 240":"7.447","plasma # 241":"9.824","plasma # 243":"6.453","plasma # 244":"7.378","plasma # 245":"10.418","plasma # 246":"9.257","plasma # 247":"13.029","plasma # 248":"10.736","plasma # 249":"17.962","plasma # 25":"8.585","plasma # 251":"7.807","plasma # 252":"10.307","plasma # 253":"15.199","plasma # 254":"10.542","plasma # 255":"7.516","plasma # 256":"9.271","plasma # 257":"8.415","plasma # 258":"15.613","plasma # 259":"10.929","plasma # 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}
}