#METABOLOMICS WORKBENCH Metadata - 2013-14 Lengyel 18 hour NIH WCMC Pilot - Newman DATATRACK_ID:143 STUDY_ID:ST000096 ANALYSIS_ID:AN000154 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 Endocannabinoid 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 Endocannabinoid 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 S29C S30C S31C S32C S33C S34C S35C S29B S30B S31B S32B S33B S34B S35B 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: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: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:Media blank | Timepoint:NA 11(12)-EpETre EA 15-HETE EA 1-AG b 11.0000 8.2000 14.0000 3.4900 7.0000 7.8000 6.4400 4.5500 4.4800 12.8000 2.8500 10.7000 11.2000 2.7200 1.1400 3.2900 5.6000 3.8200 7.9200 8.2000 8.2800 8.3500 1-LG b 69.6000 67.6000 102.0000 68.4000 54.0000 50.8000 32.6000 35.0000 60.8000 73.2000 36.2000 118.0000 42.4000 19.4000 20.0000 1.2800 11.5000 19.2000 13.2000 11.7000 12.9000 9.8000 1-OG b 143.0000 137.0000 155.0000 94.4000 119.0000 82.8000 68.0000 84.0000 112.0000 152.0000 107.0000 196.0000 120.0000 45.2000 42.8000 17.6000 24.8000 41.6000 23.6000 26.9000 22.1000 25.6000 2-AG b 3.9600 0.8160 5.5600 0.9440 2.4900 4.8400 3.2100 12.6000 5.2800 31.3000 5.3200 6.8400 15.0000 2.2000 2.0600 1.3000 2.9600 3.9600 2.5900 3.9900 3.2800 1.7400 2-LG b 24.4000 30.9000 39.8000 12.0000 25.8000 27.4000 20.2000 63.4000 53.6000 111.0000 23.7000 48.8000 44.4000 16.0000 9.0800 5.7600 6.2400 5.4400 8.5600 2.7400 5.4000 7.1000 2-OG b 33.7000 24.4000 45.2000 15.4000 30.0000 24.6000 17.2000 125.0000 55.6000 161.0000 66.0000 75.2000 88.8000 22.8000 25.8000 19.1000 11.1000 41.6000 26.8000 14.6000 6.7200 7.1800 AEA 0.1460 0.4400 0.2090 0.0780 0.0448 0.0964 0.0652 0.0542 0.0696 0.1170 0.0216 0.0386 0.0512 0.0370 0.0311 0.0284 0.0420 0.0311 0.0504 0.0321 0.0392 0.0453 aLEA 0.0190 0.0552 0.1000 0.0222 0.0131 0.0277 0.0291 0.0130 0.0024 0.0500 0.0028 0.0043 0.0073 0.0056 0.0058 0.0040 0.0014 0.0040 0.0042 0.0032 0.0040 0.0140 DEA DHEA 0.0119 0.0744 0.0123 0.0227 0.0066 0.0130 0.0660 0.0181 0.0162 0.0246 0.0220 0.0088 0.0150 0.0130 0.0070 0.0043 0.0057 0.0040 0.0063 0.0167 0.0144 0.0128 Dihomo GLA EA LEA 0.4040 1.0800 0.8800 0.3000 0.1350 0.4000 0.1580 0.1590 0.1200 0.6960 0.0652 0.0528 0.0788 0.0524 0.0428 0.0302 0.0207 0.0448 0.0143 0.0250 0.0158 0.0338 OEA 0.3930 0.5840 0.6160 0.3870 0.1620 0.3070 0.1480 0.1600 0.0648 0.6160 0.1240 0.0630 0.1090 0.0692 0.0512 0.0312 0.0388 0.0632 0.0364 0.0408 0.0382 0.0418 PEA 0.7600 1.2700 1.2400 0.8120 0.5680 0.8840 0.5240 0.5020 0.4400 1.2800 0.4720 0.2640 0.4960 0.3170 0.3790 0.3800 0.3740 0.4240 0.3980 0.3450 0.3170 0.3940 PGD2 EA PGE2 1G PGE2 EA PGF2a 1G PGF2a EA SEA 0.8600 1.1700 0.4080 0.3800 0.2940 0.5240 0.3500 0.3300 0.3330 0.5160 0.2980 0.1590 0.1910 0.1580 0.2040 0.1740 0.1650 0.2100 0.2770 0.2070 0.1660 0.2480 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 11(12)-EpETre EA 16061183 HMDB13652 15-HETE EA 44256595 1-AG b HMDB11578 1-LG b HMDB11568 1-OG b HMDB11567 2-AG b HMDB04666 2-LG b HMDB11538 2-OG b HMDB11537 AEA 5281969 C11695 HMDB04080 aLEA 5283449 HMDB13624 DEA 2186 HMDB13626 DHEA 5881 C01227 HMDB13627 Dihomo GLA EA 5282272 C13828 HMDB13625 LEA 5283446 HMDB12252 OEA 5283454 HMDB02088 PEA 1015 C16512 HMDB02100 PGD2 EA 5283120 HMDB13629 PGE2 1G 52193688 HMDB13043 PGE2 EA 5283119 PGF2a 1G 24778485 PGF2a EA 53481911 HMDB13628 SEA 27902 HMDB13078 METABOLITES_END #END