{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST003012","ANALYSIS_ID":"AN004945","VERSION":"1","CREATED_ON":"December 15, 2023, 8:45 am"},

"PROJECT":{"PROJECT_TITLE":"A High-Fat Eucaloric Diet Induces Reprometabolic Syndrome of Obesity in Normal Weight Women","PROJECT_SUMMARY":"Objective: We examined the effects of one month of a eucaloric, high-fat (48% of calories) diet (HFD) on gonadotropin secretion in normal weight women to interrogate the role of free fatty acids and insulin in mediating the relative hypogonadotropic hypogonadism of obesity. Methods: Eighteen eumenorrheic women (BMI 18-25 kg/m2) were studied in the early follicular phase of the menstrual cycle before and after exposure to a HFD with frequent blood sampling for LH and FSH, followed by an assessment of pituitary sensitivity to GnRH. Mass spectrometrybased plasma metabolomic analysis was also performed. Paired testing and time series analysis were performed as appropriate. Results: Mean endogenous LH (unstimulated) was significantly decreased after the HFD (4.3 ±1.0 vs 3.8 ± 1.0, P<0.01); mean unstimulated FSH was not changed. Both LH (10.1 ± 1.0 vs 7.2 ± 1.0, P<0.01), and FSH (9.5 ± 1.0 vs 8.8 ± 1.0, P<0.01) response to 75 ng/kg of GnRH were reduced after the HFD. Mean LH pulse amplitude and LH interpulse interval were unaffected by the dietary exposure. Eucaloric HFD exposure did not cause weight change. Plasma metabolomics confirmed adherence with elevation of fasting free fatty acids (especially long-chain mono-, poly- and highly-unsaturated fatty acids) by the last day of the HFD. Conclusion: One-month exposure to a HFD successfully induced key reproductive and metabolic features of Reprometabolic Syndrome in normal weight women. Dietary factors may underly the gonadotrope compromise seen in obesity related subfertility and that therapeutic dietary interventions, independent of weight loss, may be possible.","INSTITUTE":"University of Colorado Denver","LABORATORY":"Lab of Angelo D''Alessandro in collaboration with lab of Nanette Santoro","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"STUDY":{"STUDY_TITLE":"A High-Fat Eucaloric Diet Induces Reprometabolic Syndrome of Obesity in Normal Weight Women - metabolomics","STUDY_SUMMARY":"Objective: We examined the effects of one month of a eucaloric, high-fat (48% of calories) diet (HFD) on gonadotropin secretion in normal weight women to interrogate the role of free fatty acids and insulin in mediating the relative hypogonadotropic hypogonadism of obesity. Methods: Eighteen eumenorrheic women (BMI 18-25 kg/m2) were studied in the early follicular phase of the menstrual cycle before and after exposure to a HFD with frequent blood sampling for LH and FSH, followed by an assessment of pituitary sensitivity to GnRH. Mass spectrometrybased plasma metabolomic analysis was also performed. Paired testing and time series analysis were performed as appropriate. Results: Mean endogenous LH (unstimulated) was significantly decreased after the HFD (4.3 ±1.0 vs 3.8 ± 1.0, P<0.01); mean unstimulated FSH was not changed. Both LH (10.1 ± 1.0 vs 7.2 ± 1.0, P<0.01), and FSH (9.5 ± 1.0 vs 8.8 ± 1.0, P<0.01) response to 75 ng/kg of GnRH were reduced after the HFD. Mean LH pulse amplitude and LH interpulse interval were unaffected by the dietary exposure. Eucaloric HFD exposure did not cause weight change. Plasma metabolomics confirmed adherence with elevation of fasting free fatty acids (especially long-chain mono-, poly- and highly-unsaturated fatty acids) by the last day of the HFD. Conclusion: One-month exposure to a HFD successfully induced key reproductive and metabolic features of Reprometabolic Syndrome in normal weight women. Dietary factors may underly the gonadotrope compromise seen in obesity related subfertility and that therapeutic dietary interventions, independent of weight loss, may be possible.","INSTITUTE":"University of Colorado Denver","LAST_NAME":"Haines","FIRST_NAME":"Julie","ADDRESS":"12801 E 17th Ave, Room 1303, Aurora, Colorado, 80045, USA","EMAIL":"julie.haines@cuanschutz.edu","PHONE":"3037243339"},

"SUBJECT":{"SUBJECT_TYPE":"Human","SUBJECT_SPECIES":"Homo sapiens","TAXONOMY_ID":"9606","GENDER":"Female"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"203",
"Sample ID":"DS1-27-02",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-02"}
},
{
"Subject ID":"205",
"Sample ID":"DS1-27-05",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-05"}
},
{
"Subject ID":"213",
"Sample ID":"DS1-27-08",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-08"}
},
{
"Subject ID":"215",
"Sample ID":"DS1-27-11",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-11"}
},
{
"Subject ID":"216",
"Sample ID":"DS1-27-14",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-14"}
},
{
"Subject ID":"221",
"Sample ID":"DS1-27-17",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-17"}
},
{
"Subject ID":"226",
"Sample ID":"DS1-27-20",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-20"}
},
{
"Subject ID":"233",
"Sample ID":"DS1-27-26",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-26"}
},
{
"Subject ID":"235",
"Sample ID":"DS1-27-29",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-29"}
},
{
"Subject ID":"246",
"Sample ID":"DS1-27-35",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-35"}
},
{
"Subject ID":"247",
"Sample ID":"DS1-27-38",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-38"}
},
{
"Subject ID":"252",
"Sample ID":"DS1-27-43",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-43"}
},
{
"Subject ID":"258",
"Sample ID":"DS1-27-46",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-46"}
},
{
"Subject ID":"253",
"Sample ID":"DS1-27-49",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-49"}
},
{
"Subject ID":"250",
"Sample ID":"DS1-27-41",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-41"}
},
{
"Subject ID":"260",
"Sample ID":"DS1-27-52",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-52"}
},
{
"Subject ID":"263",
"Sample ID":"DS1-27-54",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-54"}
},
{
"Subject ID":"264",
"Sample ID":"DS1-27-56",
"Factors":{"factor":"On-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-56"}
},
{
"Subject ID":"203",
"Sample ID":"DS1-27-03",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-03"}
},
{
"Subject ID":"205",
"Sample ID":"DS1-27-06",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-06"}
},
{
"Subject ID":"213",
"Sample ID":"DS1-27-09",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-09"}
},
{
"Subject ID":"215",
"Sample ID":"DS1-27-12",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-12"}
},
{
"Subject ID":"216",
"Sample ID":"DS1-27-15",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-15"}
},
{
"Subject ID":"221",
"Sample ID":"DS1-27-18",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-18"}
},
{
"Subject ID":"226",
"Sample ID":"DS1-27-21",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-21"}
},
{
"Subject ID":"233",
"Sample ID":"DS1-27-27",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-27"}
},
{
"Subject ID":"235",
"Sample ID":"DS1-27-30",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-30"}
},
{
"Subject ID":"246",
"Sample ID":"DS1-27-36",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-36"}
},
{
"Subject ID":"247",
"Sample ID":"DS1-27-39",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-39"}
},
{
"Subject ID":"252",
"Sample ID":"DS1-27-44",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-44"}
},
{
"Subject ID":"258",
"Sample ID":"DS1-27-47",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-47"}
},
{
"Subject ID":"253",
"Sample ID":"DS1-27-50",
"Factors":{"factor":"Post-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-50"}
},
{
"Subject ID":"203",
"Sample ID":"DS1-27-01",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-01"}
},
{
"Subject ID":"205",
"Sample ID":"DS1-27-04",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-04"}
},
{
"Subject ID":"213",
"Sample ID":"DS1-27-07",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-07"}
},
{
"Subject ID":"215",
"Sample ID":"DS1-27-10",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-10"}
},
{
"Subject ID":"216",
"Sample ID":"DS1-27-13",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-13"}
},
{
"Subject ID":"221",
"Sample ID":"DS1-27-16",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-16"}
},
{
"Subject ID":"226",
"Sample ID":"DS1-27-19",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-19"}
},
{
"Subject ID":"233",
"Sample ID":"DS1-27-25",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-25"}
},
{
"Subject ID":"235",
"Sample ID":"DS1-27-28",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-28"}
},
{
"Subject ID":"246",
"Sample ID":"DS1-27-34",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-34"}
},
{
"Subject ID":"247",
"Sample ID":"DS1-27-37",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-37"}
},
{
"Subject ID":"252",
"Sample ID":"DS1-27-42",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-42"}
},
{
"Subject ID":"258",
"Sample ID":"DS1-27-45",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-45"}
},
{
"Subject ID":"253",
"Sample ID":"DS1-27-48",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-48"}
},
{
"Subject ID":"250",
"Sample ID":"DS1-27-40",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-40"}
},
{
"Subject ID":"260",
"Sample ID":"DS1-27-51",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-51"}
},
{
"Subject ID":"263",
"Sample ID":"DS1-27-53",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-53"}
},
{
"Subject ID":"264",
"Sample ID":"DS1-27-55",
"Factors":{"factor":"Pre-diet"},
"Additional sample data":{"RAW_FILE_NAME":"DS1-27-55"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Blood samples were collected from consented study subjects and plasma isolated according to standard clinical laboratory protocols.","SAMPLE_TYPE":"Blood (plasma)"},

"TREATMENT":{"TREATMENT_SUMMARY":"This study was a clinical trial (NCT02653092) and was approved by the Colorado Multiple Institutional Review Board (COMIRB). All participants provided informed consent. Women aged 18-38 with menstrual cycles 25-35 days in length and no use of reproductive hormones within 3 months of enrollment were eligible for the study. Additional eligibility criteria included: a) no history of chronic disease affecting hormone production, metabolism, or clearance and no use of medications known to interact with reproductive hormones or insulin metabolism; b) normal screening prolactin and TSH; c) normal hemoglobin A1c; d) hemoglobin > 11 g/dl at screening; e) use of reliable non-hormonal contraception throughout the study period. Because the goal was to investigate the effects of a high-fat diet, women with a baseline dietary assessment indicative of >40% calories from fat were excluded, as were women who were unable to comply with the protocol, which involved eating only foods provided by the Colorado Clinical and Translational Sciences Institute’s (CCTSI) Nutrition Services. Because of the restrictions on their ability to consume animal or dairy fats, vegans and lactose intolerant individuals were excluded. Cycle 1 was a baseline cycle; Cycle 2 entailed consumption of the high-fat diet which was continued until frequent blood sampling was completed in the early follicular phase of Cycle 3. Cycle 3 was the immediate post-high-fat-diet cycle and Cycle 4 was a recovery cycle. Participants underwent baseline and end-of-diet assessments of gonadotropin dynamics. A 6-hour frequent blood sampling study was performed to determine endogenous LH and FSH secretion (Time 0- 341 240 minutes) and concluded with a physiologic bolus of 75 ng/kg GnRH to assess pituitary 342 responsiveness (Time 240-360 minutes). Frequent sampling studies for gonadotropin determination were 343 all performed in the early follicular phase of the menstrual cycle (cycle days 2-5). Participants were also 344 seen weekly during the high-fat diet cycle and had a blood draw to assess compliance. 345 346 High-fat diet protocol. Participants completed a food preference screening survey and a 3-day diet diary, 347 which estimated their daily caloric intake and approximate proportion of daily calories derived from fat. 348 Participants were provided with a customized eucaloric high-fat diet comprised of approximately 48% 349 calories from fat, 33% calories from carbohydrates and 19% calories from protein. Food was portioned 350 into individual meals with snacks and boxed into 3-4 days’ worth at a time. Participants picked up food 351 directly from the Clinical and Translational Research Center; when this was not possible, study staff 352 delivered food to the participant. Urinary ketones were measured at each frequent blood sampling 353 session (Siemens Healthineers, Malvern, PA). During administration of the high-fat diet, blood samples were drawn weekly for 364 metabolomic analysis to determine compliance with the protocol."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Plasma for metabolomics was obtained after an overnight fast and 20 μl was extracted in 980 μl of cold methanol:acetonitrile:water (5:3:2, v/v/v). After vortexing at 4°C for 30 min, extracts were separated from the protein pellet by centrifugation for 10 min at 18,000 g at 4°C and stored at −80°C until analysis.","PROCESSING_STORAGE_CONDITIONS":"4℃","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Positive C18","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Vanquish","COLUMN_NAME":"Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)","SOLVENT_A":"100% water; 0.1% formic acid","SOLVENT_B":"100%acetonitrile; 0.1% formic acid","FLOW_GRADIENT":"0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min 95-5% B, 3-5 min hold at 5% B","FLOW_RATE":"450 uL/min","COLUMN_TEMPERATURE":"45","SAMPLE_INJECTION":"6 uL"},

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

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"Resolution 70,000, scan range 65-900 m/z, maximum injection time 200 ms, microscans 2, automatic gain control (AGC) 3 x 10^6 ions, source voltage 4.0 kV, capillary temperature 320 C, and sheath gas 45, auxiliary gas 15, and sweep gas 0 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database."},

"MS_METABOLITE_DATA":{
"Units":"peak area",

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}

}