#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