#METABOLOMICS WORKBENCH Metadata - 2013-14 Lengyel 18 hour NIH WCMC Pilot - Newman DATATRACK_ID:143 STUDY_ID:ST000096 ANALYSIS_ID:AN000153 VERSION 1 CREATED_ON 02-08-2024 #PROJECT PR:PROJECT_TITLE A study of changes in lipid metabolism of ovarian cancer cells co-cultured with PR:PROJECT_TITLE adipocytes PR:PROJECT_TYPE timecourse study PR:PROJECT_SUMMARY A study of changes in lipid metabolism of ovarian cancer cells co-cultured with PR:PROJECT_SUMMARY adipocytestimecourse studyThis West Coast Metabolomics Center pilot and PR:PROJECT_SUMMARY feasibility project was granted to Ernst Lengyel  (University of Chicago). The PR:PROJECT_SUMMARY biology of ovarian cancer (OvCa) is clearly distinct from that of most PR:PROJECT_SUMMARY epithelial tumors, in that hematogenous metastases are rare, and ovarian tumors PR:PROJECT_SUMMARY remain confined to the peritoneal cavity. The omentum, a large pad of fat tissue PR:PROJECT_SUMMARY (20x13x3cm) covering the bowel, is the most common site of OvCa metastasis. It PR:PROJECT_SUMMARY consists primarily of adipocytes, which become the principal microenvironment PR:PROJECT_SUMMARY for the OvCa cells. The underlying hypothesis for this application is that, in PR:PROJECT_SUMMARY the presence of adipocytes, the metabolism of OvCa cells is reprogramed and PR:PROJECT_SUMMARY shifts towards lipid utilization, which provides energy that facilitates tumor PR:PROJECT_SUMMARY growth and metastasis. Preliminary results suggest that primary human omental PR:PROJECT_SUMMARY adipocytes secrete cytokines which promote the metastasis of OvCa cells to the PR:PROJECT_SUMMARY omentum and their subsequent invasion. Once metastasis has occurred, OvCa cells PR:PROJECT_SUMMARY induce lipolysis in omental adipocytes, and use the energy derived from these PR:PROJECT_SUMMARY lipids to proliferate. To study the metabolic changes in the tumor PR:PROJECT_SUMMARY microenvironment we have established a 3D organotypic culture of the human PR:PROJECT_SUMMARY omentum using primary human cells established from patient tissue. Metabolic PR:PROJECT_SUMMARY studies will be performed on adipocytes and OvCa cells individually, on PR:PROJECT_SUMMARY conditioned media and on adipocytes and OvCa cells co-cultured in our 3D model, PR:PROJECT_SUMMARY with the goal of arriving at a comprehensive analysis of primary metabolites and PR:PROJECT_SUMMARY lipids in the tumor microenvironment. In the current investigation, untargeted PR:PROJECT_SUMMARY analysis of primary metabolites and complex lipids were conducted on adipocytes PR:PROJECT_SUMMARY and OvCa cells individually, on conditioned media and on adipocytes and OvCa PR:PROJECT_SUMMARY cells co-cultured in our 3D model. Analysis of oxylipins was conducted on PR:PROJECT_SUMMARY conditioned media. To gain better understanding of the dynamic regulation of PR:PROJECT_SUMMARY metabolic pathways we will also perform metabolic flux analysis using labeled PR:PROJECT_SUMMARY cells (13C-glucose, 13C-glutamine) in the 3D culture model. The primary PR:PROJECT_SUMMARY objective of this study is to gain insight into the dynamic interactions between PR:PROJECT_SUMMARY OvCa cells and human adipocytes with the anticipation of elucidating targets of PR:PROJECT_SUMMARY therapeutic intervention. PR:INSTITUTE University of California, Davis PR:DEPARTMENT Genome and Biomedical Sciences Facility PR:LABORATORY WCMC Metabolomics Core PR:LAST_NAME Fiehn PR:FIRST_NAME Oliver PR:ADDRESS 1315 Genome and Biomedical Sciences Facility,451 Health Sciences Drive, Davis, PR:ADDRESS CA 95616 PR:EMAIL ofiehn@ucdavis.edu PR:PHONE (530) 754-8258 PR:FUNDING_SOURCE NIH U24DK097154 PR:DOI http://dx.doi.org/10.21228/M82S3K #STUDY ST:STUDY_TITLE A study of changes in lipid metabolism of ovarian cancer cells co-cultured with ST:STUDY_TITLE adipocytes: UPLC-QTRAP MS analysis ST:STUDY_TYPE Timecourse ST:STUDY_SUMMARY The study investigated the interaction between omental adipocytes and OvCa ST:STUDY_SUMMARY cells, as a follow up to preliminary data indicating this leads to reprograming ST:STUDY_SUMMARY of metabolic (especially lipid) profiles in both adipocytes and OvCa cells as ST:STUDY_SUMMARY ovarian cancer cells (OvCa) readily metastasize to the omental fat pad in the ST:STUDY_SUMMARY abdomen and stimulate the release of fatty acids. In order to mimic the ST:STUDY_SUMMARY interaction between OvCa and omental adipocytes during metastasis, a coculture ST:STUDY_SUMMARY system was used that employed OvCa cells and primary human adipocytes isolated ST:STUDY_SUMMARY from omentum. Human primary adipocytes were isolated from omental explants from ST:STUDY_SUMMARY patients undergoing surgery for benign conditions. After surgical removal, ST:STUDY_SUMMARY omental tissue was digested with collagenase I, and primary cultures of ST:STUDY_SUMMARY adipocytes were established, characterized, and incorporated into the ST:STUDY_SUMMARY co-culture. The primary adipocytes were isolated and co-cultured with the OvCa ST:STUDY_SUMMARY cell line Skov3ip1. In this current submission, the the samples will be ST:STUDY_SUMMARY collected at 4, 18 and 24 hour time points post co-culture to determine the time ST:STUDY_SUMMARY dependent effect on lipid mediators, including oxylipins and ceramides. The ST:STUDY_SUMMARY study results included in this DRCC submission were the 18 hour time point data ST:STUDY_SUMMARY for oxylipins and ceramides from targeted metabolomic analysis of lipid ST:STUDY_SUMMARY mediators performed by the Newman lab. ST:INSTITUTE University of California, Davis ST:DEPARTMENT U.S.D.A. Western Human Nutrition Research Center ST:LABORATORY Newman ST:LAST_NAME Newman ST:FIRST_NAME John ST:ADDRESS 430 W. Health Sciences Dr., Davis, CA 95616 ST:EMAIL john.newman@ars.usda.gov ST:PHONE +1-530-752-1009 ST:SUBMIT_DATE 2014-07-24 #SUBJECT SU:SUBJECT_TYPE Human cells SU:SUBJECT_SPECIES Homo sapiens SU:TAXONOMY_ID 9606 SU:CELL_STRAIN_DETAILS SKOV3ip1 SU:SUBJECT_COMMENTS p80, p85, p86, p89 SU:CELL_PASSAGE_NUMBER p80, p85, p86, p89 SU:CELL_COUNTS p80, p85, p86, p89 SU:SPECIES_GROUP Human #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Additional sample data SUBJECT_SAMPLE_FACTORS Patient_1218 S29A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p94; Patient=1218; JWN ID=LNG-22; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-22.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-22.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-22.mzML SUBJECT_SAMPLE_FACTORS Patient_1222 S30A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1222; JWN ID=LNG-16; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-16.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-16.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-16.mzML SUBJECT_SAMPLE_FACTORS Patient_1223 S31A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1223; JWN ID=LNG-12; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-12.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-12.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-12.mzML SUBJECT_SAMPLE_FACTORS Patient_1229 S32A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p98; Patient=1229; JWN ID=LNG-17; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-17.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-17.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-17.mzML SUBJECT_SAMPLE_FACTORS Patient_1230 S33A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p81; Patient=1230; JWN ID=LNG-24; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-24.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-24.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-24.mzML SUBJECT_SAMPLE_FACTORS Patient_1238 S34A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p100; Patient=1238; JWN ID=LNG-25; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-25.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-25.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-25.mzML SUBJECT_SAMPLE_FACTORS Patient_1237 S35A Sample Type:Adipocyte | Timepoint:18 hours SKOV3ip1 passage=p80; Patient=1237; JWN ID=LNG-15; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-15.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-15.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-15.mzML SUBJECT_SAMPLE_FACTORS Patient_1171 S17A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p80; Patient=#1171; JWN_ID=A4-S17A; RAW_FILE_NAME=Lengyal Pilot Oxys2-A4-S17A.mzML SUBJECT_SAMPLE_FACTORS Patient_1181 S19A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1181; JWN_ID=A7-S19A; RAW_FILE_NAME=Lengyal Pilot Oxys-A7-S19A.mzML SUBJECT_SAMPLE_FACTORS Patient_1182 S20A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1182; JWN_ID=A11-S20A; RAW_FILE_NAME=Lengyal Pilot Oxys2-A11-S20A.mzML SUBJECT_SAMPLE_FACTORS Patient_1193 S23A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1193; JWN_ID=B3-S23A; RAW_FILE_NAME=Lengyal Pilot Oxys2-B3-S23A.mzML SUBJECT_SAMPLE_FACTORS Patient_1194 S24A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p85; Patient=#1194; JWN_ID=B6-S24A; RAW_FILE_NAME=Lengyal Pilot Oxys-B6-S24A.mzML SUBJECT_SAMPLE_FACTORS Patient_1197 S25A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1197; JWN_ID=B9-S25A; RAW_FILE_NAME=Lengyal Pilot Oxys2-B9-S25A.mzML SUBJECT_SAMPLE_FACTORS Patient_1198 S26A Sample Type:Adipocyte | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1198; JWN_ID=B12-S26A; RAW_FILE_NAME=Lengyal Pilot Oxys2-B12-S26A.mzML SUBJECT_SAMPLE_FACTORS Patient_1218 S29C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p94; Patient=1218; JWN ID=LNG-18 LNG-19; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-18.mzML Lengyal 18 hr Pilot Oxys r-LNG-19.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-18.mzML Lengyal 18 hr Pilot Endos-LNG-19.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-18.mzML Lengyal 18 hr Pilot Cer-LNG-19.mzML SUBJECT_SAMPLE_FACTORS Patient_1222 S30C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1222; JWN ID=LNG-14; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-14.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-14.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-14.mzML SUBJECT_SAMPLE_FACTORS Patient_1223 S31C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1223; JWN ID=LNG-21; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-21.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-21.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-21.mzML SUBJECT_SAMPLE_FACTORS Patient_1229 S32C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p98; Patient=1229; JWN ID=LNG-28; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-28.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-28.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-28.mzML SUBJECT_SAMPLE_FACTORS Patient_1230 S33C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p81; Patient=1230; JWN ID=LNG-9 LNG-10; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-9.mzML Lengyal 18 hr Pilot Oxys r-LNG-10.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-9.mzML Lengyal 18 hr Pilot Endos-LNG-10.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-9.mzML Lengyal 18 hr Pilot Cer-LNG-10.mzML SUBJECT_SAMPLE_FACTORS Patient_1238 S34C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p100; Patient=1238; JWN ID=LNG-6; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-6.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-6.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-6.mzML SUBJECT_SAMPLE_FACTORS Patient_1237 S35C Sample Type:Co-culture | Timepoint:18 hours SKOV3ip1 passage=p80; Patient=1237; JWN ID=LNG-27; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-27.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-27.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-27.mzML SUBJECT_SAMPLE_FACTORS Patient_1171 S17C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p80; Patient=#1171; JWN_ID=A6-S17C; RAW_FILE_NAME=Lengyal Pilot Oxys2-A6-S17C.mzML SUBJECT_SAMPLE_FACTORS Patient_1181 S19C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1181; JWN_ID=A9-S19C-1; RAW_FILE_NAME=Lengyal Pilot Oxys-A9-S19C-1.mzML SUBJECT_SAMPLE_FACTORS Patient_1182 S20C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1182; JWN_ID=A13-S20C-1; RAW_FILE_NAME=Lengyal Pilot Oxys2-A13-S20-C-1.mzML SUBJECT_SAMPLE_FACTORS Patient_1193 S23C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1193; JWN_ID=B5-S23C; RAW_FILE_NAME=Lengyal Pilot Oxys2-B5-S23C.mzML SUBJECT_SAMPLE_FACTORS Patient_1194 S24C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p85; Patient=#1194; JWN_ID=B8-S24C; RAW_FILE_NAME=Lengyal Pilot Oxys-B8-S24C.mzML SUBJECT_SAMPLE_FACTORS Patient_1197 S25C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1197; JWN_ID=B11-S25C; RAW_FILE_NAME=Lengyal Pilot Oxys2-B11-S25C.mzML SUBJECT_SAMPLE_FACTORS Patient_1198 S26C Sample Type:Co-culture | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1198; JWN_ID=B14-S26C-1; RAW_FILE_NAME=Lengyal Pilot Oxys2-B14-S26C-1.mzML SUBJECT_SAMPLE_FACTORS Patient_1218 S29B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p94; Patient=1218; JWN ID=LNG-20; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-20.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-20.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-20.mzML SUBJECT_SAMPLE_FACTORS Patient_1222 S30B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1222; JWN ID=LNG-23; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-23.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-23.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-23.mzML SUBJECT_SAMPLE_FACTORS Patient_1223 S31B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p96; Patient=1223; JWN ID=LNG-13; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-13.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-13.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-13.mzML SUBJECT_SAMPLE_FACTORS Patient_1229 S32B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p98; Patient=1229; JWN ID=LNG-26; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-26.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-26.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-26.mzML SUBJECT_SAMPLE_FACTORS Patient_1230 S33B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p81; Patient=1230; JWN ID=LNG-11; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-11.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-11.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-11.mzML SUBJECT_SAMPLE_FACTORS Patient_1238 S34B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p100; Patient=1238; JWN ID=LNG-7; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-7.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-7.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-7.mzML SUBJECT_SAMPLE_FACTORS Patient_1237 S35B Sample Type:Control | Timepoint:18 hours SKOV3ip1 passage=p80; Patient=1237; JWN ID=LNG-8; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-8.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-8.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-8.mzML SUBJECT_SAMPLE_FACTORS Patient_1171 S17B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p80; Patient=#1171; JWN_ID=A5-S17B; RAW_FILE_NAME=Lengyal Pilot Oxys-A5-S17B.mzML SUBJECT_SAMPLE_FACTORS Patient_1181 S19B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1181; JWN_ID=A8-S19B; RAW_FILE_NAME=Lengyal Pilot Oxys2-A8-S19B.mzML SUBJECT_SAMPLE_FACTORS Patient_1182 S20B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p86; Patient=#1182; JWN_ID=A12-S20B; RAW_FILE_NAME=Lengyal Pilot Oxys2-A12-S20B.mzML SUBJECT_SAMPLE_FACTORS Patient_1193 S23B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1193; JWN_ID=B4-S23B; RAW_FILE_NAME=Lengyal Pilot Oxys-B4-S23B.mzML SUBJECT_SAMPLE_FACTORS Patient_1194 S24B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p85; Patient=#1194; JWN_ID=B7-S24B; RAW_FILE_NAME=Lengyal Pilot Oxys2-B7-S24B.mzML SUBJECT_SAMPLE_FACTORS Patient_1197 S25B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1197; JWN_ID=B10-S25B; RAW_FILE_NAME=Lengyal Pilot Oxys2-B10-S25B.mzML SUBJECT_SAMPLE_FACTORS Patient_1198 S26B Sample Type:Control | Timepoint:4 hours SKOV3ip1 passage=p89; Patient=#1198; JWN_ID=B13-S26B; RAW_FILE_NAME=Lengyal Pilot Oxys2-B13-S26B.mzML SUBJECT_SAMPLE_FACTORS - Media blank Sample Type:Media blank | Timepoint:4 hours SKOV3ip1 passage=Media blank; Patient=Media blank; JWN_ID=Media blank; RAW_FILE_NAME=Lengyal Pilot Oxys-A3-Blank Med.mzML SUBJECT_SAMPLE_FACTORS - Blank media Sample Type:Media blank | Timepoint:NA SKOV3ip1 passage=-; Patient=-; JWN ID=LNG-3 LNG-4 LNG-5; RAW_FILE_NAME=(Oxylipin data)Lengyal 18 hr Pilot Oxys r-LNG-3.mzML Lengyal 18 hr Pilot Oxys r-LNG-4.mzML Lengyal 18 hr Pilot Oxys r-LNG-5.mzML; File name (Ceramide data)=Lengyal 18 hr Pilot Endos-LNG-3.mzML Lengyal 18 hr Pilot Endos-LNG-4.mzML Lengyal 18 hr Pilot Endos-LNG-5.mzML; File name (Endocannabinoid data)=Lengyal 18 hr Pilot Cer-LNG-3.mzML Lengyal 18 hr Pilot Cer-LNG-4.mzML Lengyal 18 hr Pilot Cer-LNG-5.mzML #COLLECTION CO:COLLECTION_SUMMARY Samples were collected at 18 hours post co-culture CO:COLLECTION_PROTOCOL_FILENAME Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf CO:SAMPLE_TYPE Media CO:STORAGE_CONDITIONS -80 C #TREATMENT TR:TREATMENT_SUMMARY The study investigated the interaction between omental adipocytes and OvCa TR:TREATMENT_SUMMARY cells, as a follow up to preliminary data indicating this leads to reprograming TR:TREATMENT_SUMMARY of metabolic (especially lipid) profiles in both adipocytes and OvCa cells as TR:TREATMENT_SUMMARY ovarian cancer cells (OvCa) readily metastasize to the omental fat pad in the TR:TREATMENT_SUMMARY abdomen and stimulate the release of fatty acids. In order to mimic the TR:TREATMENT_SUMMARY interaction between OvCa and omental adipocytes during metastasis, a coculture TR:TREATMENT_SUMMARY system was used that employed OvCa cells and primary human adipocytes isolated TR:TREATMENT_SUMMARY from omentum. Human primary adipocytes were isolated from omental explants from TR:TREATMENT_SUMMARY patients undergoing surgery for benign conditions. After surgical removal, TR:TREATMENT_SUMMARY omental tissue was digested with collagenase I, and primary cultures of TR:TREATMENT_SUMMARY adipocytes were established, characterized, and incorporated into the TR:TREATMENT_SUMMARY co-culture. The primary adipocytes were isolated and co-cultured with the OvCa TR:TREATMENT_SUMMARY cell line Skov3ip1. In this current submission, the the samples will be TR:TREATMENT_SUMMARY collected at 4, 18 and 24 hour time points post co-culture to determine the time TR:TREATMENT_SUMMARY dependent effect on lipid mediators, including oxylipins and ceramides. The TR:TREATMENT_SUMMARY study results included in this DRCC submission were the 18 hour time point data TR:TREATMENT_SUMMARY for oxylipins and ceramides. TR:TREATMENT_PROTOCOL_FILENAME Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf TR:CELL_STORAGE -80 C #SAMPLEPREP SP:SAMPLEPREP_SUMMARY See sample prep protocol file SP:SAMPLEPREP_PROTOCOL_FILENAME Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Enpdfannabinoid_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf;Newman_Lab_Ceramide_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf SP:PROCESSING_STORAGE_CONDITIONS - 20 °C SP:EXTRACTION_METHOD SPE SP:EXTRACT_CONCENTRATION_DILUTION 250 µL SP:EXTRACT_CLEANUP SPE SP:EXTRACT_STORAGE - 20 °C SP:SAMPLE_RESUSPENSION 100 µL SP:SAMPLE_SPIKING See sample prep protocol file #CHROMATOGRAPHY CH:CHROMATOGRAPHY_SUMMARY Oxylipin analysis CH:METHODS_FILENAME Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_18hr.pdf;Newman_Lab_Oxylipin_Extraction_&_Analysis_Protocol_Lengyel_WCMC_P&F_4hr.pdf CH:COLUMN_TEMPERATURE 60 C CH:FLOW_GRADIENT See protocol/methods file CH:FLOW_RATE 0.25 CH:INTERNAL_STANDARD See protocol/methods file CH:RETENTION_TIME See protocol/methods file CH:SAMPLE_INJECTION 5 L CH:SOLVENT_A 100% water; 0.1% acetic acid CH:SOLVENT_B 90% acetonitrile/ 10% isopropanol CH:ANALYTICAL_TIME 16 min CH:WEAK_WASH_SOLVENT_NAME 20% methanol, 10% isopropanol CH:WEAK_WASH_VOLUME 600 L CH:STRONG_WASH_SOLVENT_NAME 50:50 Acetonitrile:Methanol CH:STRONG_WASH_VOLUME 600 L CH:SAMPLE_LOOP_SIZE 17 L #ANALYSIS AN:LABORATORY_NAME Newman AN:ANALYSIS_TYPE MS AN:ANALYSIS_COMMENTS Ceramide analysis AN:ACQUISITION_DATE 41732 AN:ACQUISITION_TIME 15 min AN:SOFTWARE_VERSION AB Sciex Analyst version 1.6.2 AN:OPERATOR_NAME Michael La Frano AN:DETECTOR_TYPE API 4000 QTrap (AB Sciex, Framingham, MA, USA) AN:DATA_FORMAT .mzML #MS MS:INSTRUMENT_NAME ABI Sciex API 4000 QTrap MS:INSTRUMENT_TYPE Triple quadrupole MS:MS_TYPE ESI MS:MS_COMMENTS Ceramide analysis MS:ION_MODE POSITIVE MS:ION_SOURCE_TEMPERATURE See protocol/methods file MS:ION_SPRAY_VOLTAGE See protocol/methods file #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS nM MS_METABOLITE_DATA_START Samples S29A S30A S31A S32A S33A S34A S35A S17A S19A S20A S23A S24A S25A S26A S29C S30C S31C S32C S33C S34C S35C S17C S19C S20C S23C S24C S25C S26C S29B S30B S31B S32B S33B S34B S35B S17B S19B S20B S23B S24B S25B S26B Media blank Blank media Factors Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:18 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Adipocyte | Timepoint:4 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:18 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Co-culture | Timepoint:4 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:18 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Control | Timepoint:4 hours Sample Type:Media blank | Timepoint:4 hours Sample Type:Media blank | Timepoint:NA 18:0 Sphingosine-1P 18:1 Sphingosine 5.2000 9.4800 4.8800 3.3300 7.3200 15.7000 12.6000 1.0640 2.6400 2.7600 2.4560 3.2560 2.1840 2.8360 10.1000 5.4800 6.2800 5.9200 4.0200 6.4800 8.7600 4.2000 3.1860 2.1900 1.9800 3.5320 3.2520 1.6700 2.8800 3.5100 2.6600 3.1500 3.3000 2.0900 2.1400 1.2440 2.2840 0.8280 0.9560 1.7120 0.8120 1.2000 0.5680 3.2600 18:1 Sphingosine-1P 0.2970 0.1730 1.2200 0.3530 0.1820 0.3060 0.2580 0.5115 0.5560 0.5280 0.1600 0.5880 1.1500 0.0852 0.4160 0.4880 0.7480 0.4560 0.9320 0.9160 0.6520 2.1867 C14 Ceramide 2.7400 1.1200 1.9800 1.1800 2.0800 4.5200 1.6100 0.4160 1.1960 0.7840 0.7280 1.7720 0.8280 3.0200 4.4400 1.8900 4.4000 2.3700 2.5900 4.1600 2.3700 0.8080 0.9480 0.5760 1.0760 1.6400 2.0320 1.7100 0.3040 0.4200 0.2250 0.2720 0.2680 0.2360 0.1680 0.0968 0.1088 0.0744 0.0972 0.0632 0.0588 0.1120 0.0251 0.1530 C16 Ceramide 46.8000 26.5000 32.7000 13.4000 35.2000 114.0000 31.8000 6.7200 20.3200 11.7600 17.0400 40.8000 17.9200 37.4800 90.4000 46.4000 90.8000 35.2000 41.8000 94.8000 44.0000 13.2400 16.2400 7.4400 19.5600 36.2000 43.6000 25.4200 8.8400 13.5000 5.4000 7.2400 7.7200 6.0000 5.2400 1.8040 1.7920 1.5200 1.9040 1.0440 0.9320 2.4120 0.6960 3.0400 C18:1 Ceramide 1.5500 1.5000 1.3900 0.8760 1.0800 4.0800 1.8700 0.3356 1.5120 1.0640 0.9440 2.6560 0.5520 1.7080 2.6100 1.9900 4.3600 1.9300 1.2500 2.9800 2.3200 0.6200 1.2400 0.5600 1.0000 2.8640 1.2720 1.0860 0.4200 0.5960 0.2850 0.4080 0.2880 0.2880 0.1190 0.0612 0.0102 0.0616 0.0824 0.0305 0.0246 0.0688 0.0336 0.1490 C18 Ceramide 4.2400 3.5700 4.1200 1.0000 2.7500 13.2000 3.8200 0.9480 5.0000 2.3080 1.9720 6.7200 1.9000 5.7200 9.2600 6.0800 12.1000 5.0000 3.9900 11.3000 5.0800 1.4960 4.0500 2.0980 2.8040 7.8800 4.8800 4.0160 1.2600 2.0500 0.4440 1.1200 1.1400 0.9200 0.7280 0.2556 0.2472 0.2768 0.2992 0.3560 0.3248 0.5800 0.1536 0.4180 C18 dihydroceramide 0.1040 0.1000 0.0860 1.0700 0.1572 0.6400 0.2844 0.1328 0.7960 0.3072 0.7360 0.6040 0.2010 0.7120 0.0376 0.1210 1.1700 0.1480 0.2216 0.3896 0.1614 0.2728 0.8040 0.7440 0.5120 0.0319 0.0956 0.0536 0.0936 0.0820 0.0940 0.0516 0.0488 C20 Ceramide 3.6000 3.2700 2.9700 1.4000 2.6100 13.0000 5.1200 1.1920 4.0400 2.2640 2.7240 3.4800 2.0200 4.2000 6.3600 3.6800 6.9200 2.6000 3.0800 8.2800 4.9600 1.6200 3.3380 1.6800 3.1640 5.2000 5.0400 2.7440 0.1740 0.4160 0.1290 0.4200 0.1190 0.1860 0.1240 0.1168 0.1416 0.1648 0.1020 0.1552 0.1252 0.3256 0.0704 0.0819 C24 Ceramide 16.5000 11.1000 6.7200 5.5200 16.4000 34.1000 18.8000 2.3840 6.3200 9.7600 5.5600 11.7600 9.0400 8.9200 40.8000 15.1000 21.0000 19.5000 20.4000 22.6000 20.6000 7.7200 6.2800 4.6000 8.9600 13.7600 22.3600 8.7400 2.7900 4.2000 1.4600 3.3500 1.7100 1.9800 1.0600 0.4400 0.4120 0.5000 0.4200 0.4800 0.4640 0.7480 0.1820 C24 dihydroceramide 0.2010 0.1250 0.0370 0.1610 0.0028 0.4480 0.1460 0.6100 0.0896 0.1620 0.3280 0.1350 0.1720 0.1560 0.0528 0.0049 0.1360 0.0221 0.0532 0.0673 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name pubchem_id inchi_key kegg_id other_id other_id_type ri ri_type moverz_quant 18:0 Sphingosine-1P 644260 C01120 HMDB01383 UCDavis_Newman_ID 18:1 Sphingosine 42608348 C00319 HMDB00252 UCDavis_Newman_ID 18:1 Sphingosine-1P 5283560 C06124 HMDB00277 UCDavis_Newman_ID C14 Ceramide 5282310 C13916 C16 Ceramide 5283564 HMDB04949 UCDavis_Newman_ID C18:1 Ceramide 5283563 HMDB04948 UCDavis_Newman_ID C18 Ceramide 5283565 C00195 HMDB04950 UCDavis_Newman_ID C18 dihydroceramide 5283573 C20 Ceramide 5283566 HMDB04951 UCDavis_Newman_ID C24 Ceramide 5283571 HMDB04956 UCDavis_Newman_ID C24 dihydroceramide 5283577 HMDB11768 UCDavis_Newman_ID METABOLITES_END #END