{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000956","ANALYSIS_ID":"AN001568","VERSION":"1","CREATED_ON":"April 19, 2018, 12:26 pm"},

"PROJECT":{"PROJECT_TITLE":"Determine metabolomics signatures important for the ability of Streptococus gallolyticus to promote colon cancer cell proliferation","PROJECT_SUMMARY":"It is now known that intestinal microbiota influences the development of colorectal cancer (CRC). This microbe-CRC connection suggests a potential paradigm shift in the way CRC is detected, treated and managed. Knowledge of specific microbial components involved in the development of CRC is critical to moving this field forward. Among the bacterial species known to associate with CRC, Streptococcus gallolyticus subsp. gallolyticus (Sg), previously known as S. bovis biotype I, stands out as having a strong and well-documented clinical association supported by numerous case reports and surveys over the past several decades. We and others also found that Sg is present in a substantial percentage of CRC patients (up to ~ 74%). We further demonstrated that Sg actively promotes colon tumor growth. These exciting discoveries underscore the importance of Sg in CRC with respect to both function and clinical relevance. Further investigation into the molecular details of the Sg-CRC relationship should have a high priority. Going forward, the key question is how Sg promotes colon tumor development. Data from our lab led us to hypothesize that Sg produces certain metabolites that contribute to its ability to promote cell proliferation. We propose to identify the metabolites important for promoting colon cancer cell proliferation. Our approach is based on two recent findings. We discovered that there are variations among Sg strains in the ability to stimulate host cell proliferation. We also observed that the ability of Sg to promote cell proliferation is bacterial growth phase regulated. Thus by comparing the metabolomics profiles of different Sg strains, and Sg strains from different growth phase co-cultured with colon cancer cells, respectively, we will identify metabolomics signatures that correlate with the ability of Sg to promote cell proliferation. These metabolites will then be investigated in more detail in future studies. In addition, DNA methylation pattern in cells treated with Sg, negative control bacteria and media only will also be compared.","INSTITUTE":"Texas A&M Health Scicence Center, Institute of Biosciences and Technology","DEPARTMENT":"Center for Infectious and Inflammatory Diseases","LAST_NAME":"Xu","FIRST_NAME":"Yi","ADDRESS":"2121 W. Holcombe Blvd., Houston, TX 77030","EMAIL":"yxu@ibt.tamhsc.edu","PHONE":"NA"},

"STUDY":{"STUDY_TITLE":"Determine metabolomics signatures important for the ability of Streptococus gallolyticus to promote colon cancer cell proliferation","STUDY_SUMMARY":"HT29 cells were treated with Sg TX20005 stationary phase, Sg TX20005 exponential phase (negative control), and Sg TX20008 stationary phase (negative control), and media only (baseline control). The cells were washed and aliquoted. One aliquot was pelleted and stored at -80C for metabolomics analysis. DNA was also extracted from the other aliquot for DNA methylation studies.","INSTITUTE":"University of Florida","DEPARTMENT":"SECIM","LAST_NAME":"Xu","FIRST_NAME":"Yi","ADDRESS":"2121 W. Holcombe Blvd., Houston, TX 77030","EMAIL":"yxu@ibt.tamhsc.edu","PHONE":"NA","NUM_GROUPS":"4","TOTAL_SUBJECTS":"40","STUDY_COMMENTS":"SECIM pilot and feasibility, NIH U24 DK097209"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","GENOTYPE_STRAIN":"HT29 cell line (colorectal cancer)"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"HT29-1",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-2",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-3",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-4",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-5",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-6",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-7",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-8",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-9",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-10",
"Factors":{"Treatment":"Control"},
"Additional sample data":{"Label":"No bacteria","Description":"media only (baseline control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-1",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-2",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-3",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-4",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-5",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-6",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-7",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-8",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-9",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX08-10",
"Factors":{"Treatment":"Condition-1"},
"Additional sample data":{"Label":"TX20008-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-1",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-2",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-3",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-4",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-5",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-6",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-7",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-8",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-9",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-S-10",
"Factors":{"Treatment":"Condition-2"},
"Additional sample data":{"Label":"TX20005-S","Description":"stationary phase"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-1",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-2",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-3",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-4",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-5",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-6",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-7",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-8",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-9",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
},
{
"Subject ID":"-",
"Sample ID":"HT29-TX05-E-10",
"Factors":{"Treatment":"Condition-3"},
"Additional sample data":{"Label":"TX20005-E","Description":"exponential phase (negative control)"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"HT29 cells were treated with Sg TX20005 stationary phase, Sg TX20005 exponential phase (negative control), and Sg TX20008 stationary phase (negative control), and media only (baseline control). The cells were washed and aliquoted. One aliquot was pelleted and stored at -80C for metabolomics analysis. DNA was also extracted from the other aliquot for DNA methylation studies.","SAMPLE_TYPE":"new"},

"TREATMENT":{"TREATMENT_SUMMARY":"HT29 cells were co-cultured with S. streptococcus gallolyticus strains TX20008 (stationary phase), TX20005 (Stationary phase) and TX20005 (Exponential phase). Cell storage: After collection ells were stored at -80 C Cell growth container:10 cm plates Cell percent confluence:80% Cell media: F12/DMEM 1:1 + 10% FBS Cell harvesting: Trypisinization"},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"No details provided. HT29 cells","SAMPLEPREP_PROTOCOL_FILENAME":"GMetabolomics_LCMS_Protocol_092117.pdf;Appendix_A_Internal_Standard_Prep_GLCMS.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Dionex Ultimate 3000","COLUMN_NAME":"ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)","FLOW_RATE":"350ul/min","SOLVENT_A":"0.1% Formic Acid in Water","SOLVENT_B":"Acetonitrile"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"MS_COMMENTS":"-","INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_RESULTS_FILE":"ST000956_AN001568_Results.txt UNITS:peak height"},

"MS_METABOLITE_DATA":{
"Units":"peak height",

"Data":[{"Metabolite":"(R)-MALATE","HT29-1":"851631","HT29-2":"167408","HT29-3":"338413","HT29-4":"186106","HT29-5":"169486","HT29-6":"679833","HT29-7":"516554","HT29-8":"545132","HT29-9":"114488","HT29-10":"326595","HT29-TX08-1":"261687","HT29-TX08-4":"150779","HT29-TX08-2":"101328","HT29-TX08-5":"171118","HT29-TX08-3":"79853","HT29-TX08-6":"125469","HT29-TX08-7":"153357","HT29-TX08-8":"543248","HT29-TX08-9":"590008","HT29-TX08-10":"449426","HT29-TX05-S-3":"820085","HT29-TX05-S-1":"1239402","HT29-TX05-S-2":"313955","HT29-TX05-S-4":"1071899","HT29-TX05-S-6":"534838","HT29-TX05-S-5":"994998","HT29-TX05-S-8":"572417","HT29-TX05-S-9":"885229","HT29-TX05-E-1":"251027","HT29-TX05-S-10":"1063298","HT29-TX05-S-7":"952893","HT29-TX05-E-2":"484745","HT29-TX05-E-3":"316908","HT29-TX05-E-4":"99984","HT29-TX05-E-6":"238462","HT29-TX05-E-7":"107651","HT29-TX05-E-5":"228948","HT29-TX05-E-8":"809546","HT29-TX05-E-9":"761388","HT29-TX05-E-10":"771542"},{"Metabolite":"2-hydroxyglutarate+Na","HT29-1":"17669","HT29-2":"4689","HT29-3":"6039","HT29-4":"10725","HT29-5":"11895","HT29-6":"15187","HT29-7":"13660","HT29-8":"16015","HT29-9":"6307","HT29-10":"6247","HT29-TX08-1":"11303","HT29-TX08-4":"13504","HT29-TX08-2":"5505","HT29-TX08-5":"13124","HT29-TX08-3":"5388","HT29-TX08-6":"9377","HT29-TX08-7":"10440","HT29-TX08-8":"20536","HT29-TX08-9":"17440","HT29-TX08-10":"11672","HT29-TX05-S-3":"17765","HT29-TX05-S-1":"21066","HT29-TX05-S-2":"15874","HT29-TX05-S-4":"21453","HT29-TX05-S-6":"17910","HT29-TX05-S-5":"24468","HT29-TX05-S-8":"18784","HT29-TX05-S-9":"20491","HT29-TX05-E-1":"5887","HT29-TX05-S-10":"20269","HT29-TX05-S-7":"33522","HT29-TX05-E-2":"13156","HT29-TX05-E-3":"12301","HT29-TX05-E-4":"7276","HT29-TX05-E-6":"5520","HT29-TX05-E-7":"10846","HT29-TX05-E-5":"12723","HT29-TX05-E-8":"21692","HT29-TX05-E-9":"25482","HT29-TX05-E-10":"21721"},{"Metabolite":"2-Hydroxyphenylalanine;L-TYROSINE","HT29-1":"21800000","HT29-2":"13400000","HT29-3":"14200000","HT29-4":"13100000","HT29-5":"13500000","HT29-6":"20500000","HT29-7":"17800000","HT29-8":"21000000","HT29-9":"10500000","HT29-10":"16600000","HT29-TX08-1":"13500000","HT29-TX08-4":"10600000","HT29-TX08-2":"10500000","HT29-TX08-5":"12600000","HT29-TX08-3":"10900000","HT29-TX08-6":"12900000","HT29-TX08-7":"15800000","HT29-TX08-8":"18000000","HT29-TX08-9":"20200000","HT29-TX08-10":"17200000","HT29-TX05-S-3":"20900000","HT29-TX05-S-1":"27700000","HT29-TX05-S-2":"13900000","HT29-TX05-S-4":"24700000","HT29-TX05-S-6":"13000000","HT29-TX05-S-5":"23800000","HT29-TX05-S-8":"16900000","HT29-TX05-S-9":"23000000","HT29-TX05-E-1":"13500000","HT29-TX05-S-10":"24400000","HT29-TX05-S-7":"20900000","HT29-TX05-E-2":"18400000","HT29-TX05-E-3":"17000000","HT29-TX05-E-4":"10400000","HT29-TX05-E-6":"14100000","HT29-TX05-E-7":"9779960","HT29-TX05-E-5":"14400000","HT29-TX05-E-8":"22900000","HT29-TX05-E-9":"21100000","HT29-TX05-E-10":"21600000"},{"Metabolite":"3'-CMP","HT29-1":"15283","HT29-2":"5926","HT29-3":"22185","HT29-4":"24756","HT29-5":"20342","HT29-6":"14974","HT29-7":"27653","HT29-8":"18122","HT29-9":"33667","HT29-10":"27746","HT29-TX08-1":"38077","HT29-TX08-4":"12148","HT29-TX08-2":"33527","HT29-TX08-5":"27463","HT29-TX08-3":"29931","HT29-TX08-6":"27443","HT29-TX08-7":"43838","HT29-TX08-8":"20594","HT29-TX08-9":"19956","HT29-TX08-10":"20353","HT29-TX05-S-3":"23783","HT29-TX05-S-1":"37813","HT29-TX05-S-2":"104366","HT29-TX05-S-4":"30052","HT29-TX05-S-6":"80055","HT29-TX05-S-5":"44203","HT29-TX05-S-8":"109141","HT29-TX05-S-9":"32344","HT29-TX05-E-1":"35619","HT29-TX05-S-10":"38573","HT29-TX05-S-7":"112265","HT29-TX05-E-2":"28451","HT29-TX05-E-3":"30077","HT29-TX05-E-4":"32758","HT29-TX05-E-6":"50318","HT29-TX05-E-7":"37543","HT29-TX05-E-5":"41664","HT29-TX05-E-8":"27335","HT29-TX05-E-9":"25262","HT29-TX05-E-10":"25852"},{"Metabolite":"3-UREIDOPROPIONATE","HT29-1":"64757","HT29-2":"38885","HT29-3":"69837","HT29-4":"100678","HT29-5":"57879","HT29-6":"80123","HT29-7":"65753","HT29-8":"49583","HT29-9":"98740","HT29-10":"108328","HT29-TX08-1":"76609","HT29-TX08-4":"25555","HT29-TX08-2":"44157","HT29-TX08-5":"41948","HT29-TX08-3":"72422","HT29-TX08-6":"54014","HT29-TX08-7":"107087","HT29-TX08-8":"36056","HT29-TX08-9":"76400","HT29-TX08-10":"42595","HT29-TX05-S-3":"55270","HT29-TX05-S-1":"106602","HT29-TX05-S-2":"59787","HT29-TX05-S-4":"95763","HT29-TX05-S-6":"51515","HT29-TX05-S-5":"93448","HT29-TX05-S-8":"57231","HT29-TX05-S-9":"71382","HT29-TX05-E-1":"94437","HT29-TX05-S-10":"76949","HT29-TX05-S-7":"110182","HT29-TX05-E-2":"61485","HT29-TX05-E-3":"124535","HT29-TX05-E-4":"67536","HT29-TX05-E-6":"118203","HT29-TX05-E-7":"58715","HT29-TX05-E-5":"71576","HT29-TX05-E-8":"77576","HT29-TX05-E-9":"83930","HT29-TX05-E-10":"75897"},{"Metabolite":"4-AMINOBUTANOATE 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