Summary of Study ST003012

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001875. The data can be accessed directly via it's Project DOI: 10.21228/M8BX4T This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST003012
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
Submit Date	2023-12-14
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2024-01-02
Release Version	1

Select appropriate tab below to view additional metadata details:

Project:

Project ID:	PR001875
Project DOI:	doi: 10.21228/M8BX4T
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

Subject:

Subject ID:	SU003126
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Gender:	Female

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id	local_sample_id	factor
SA327071	DS1-27-49	On-diet
SA327072	DS1-27-02	On-diet
SA327073	DS1-27-43	On-diet
SA327074	DS1-27-41	On-diet
SA327075	DS1-27-52	On-diet
SA327076	DS1-27-56	On-diet
SA327077	DS1-27-54	On-diet
SA327078	DS1-27-38	On-diet
SA327079	DS1-27-46	On-diet
SA327080	DS1-27-11	On-diet
SA327081	DS1-27-35	On-diet
SA327082	DS1-27-05	On-diet
SA327083	DS1-27-14	On-diet
SA327084	DS1-27-08	On-diet
SA327085	DS1-27-17	On-diet
SA327086	DS1-27-26	On-diet
SA327087	DS1-27-29	On-diet
SA327088	DS1-27-20	On-diet
SA327089	DS1-27-36	Post-diet
SA327090	DS1-27-30	Post-diet
SA327091	DS1-27-39	Post-diet
SA327092	DS1-27-50	Post-diet
SA327093	DS1-27-47	Post-diet
SA327094	DS1-27-44	Post-diet
SA327095	DS1-27-27	Post-diet
SA327096	DS1-27-03	Post-diet
SA327097	DS1-27-21	Post-diet
SA327098	DS1-27-06	Post-diet
SA327099	DS1-27-09	Post-diet
SA327100	DS1-27-18	Post-diet
SA327101	DS1-27-15	Post-diet
SA327102	DS1-27-12	Post-diet
SA327103	DS1-27-45	Pre-diet
SA327104	DS1-27-42	Pre-diet
SA327105	DS1-27-48	Pre-diet
SA327106	DS1-27-55	Pre-diet
SA327107	DS1-27-37	Pre-diet
SA327108	DS1-27-53	Pre-diet
SA327109	DS1-27-51	Pre-diet
SA327110	DS1-27-40	Pre-diet
SA327111	DS1-27-19	Pre-diet
SA327112	DS1-27-07	Pre-diet
SA327113	DS1-27-04	Pre-diet
SA327114	DS1-27-01	Pre-diet
SA327115	DS1-27-10	Pre-diet
SA327116	DS1-27-13	Pre-diet
SA327117	DS1-27-28	Pre-diet
SA327118	DS1-27-25	Pre-diet
SA327119	DS1-27-16	Pre-diet
SA327120	DS1-27-34	Pre-diet

Showing results 1 to 50 of 50

Showing results 1 to 50 of 50

Collection:

Collection ID:	CO003119
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 ID:	TR003135
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.

Treatment ID:

TR003135

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.

Sample Preparation:

Sampleprep ID:	SP003132
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℃

Combined analysis:

Analysis ID	AN004944	AN004945
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Thermo Vanquish	Thermo Vanquish
Column	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
MS Type	ESI	ESI
MS instrument type	Orbitrap	Orbitrap
MS instrument name	Thermo Q Exactive Orbitrap	Thermo Q Exactive Orbitrap
Ion Mode	NEGATIVE	POSITIVE
Units	peak area	peak area

Chromatography:

Chromatography ID:	CH003732
Chromatography Summary:	Negative C18
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
Column Temperature:	45
Flow Gradient:	0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min 100-0% B, 3-5 min hold at 0% B
Flow Rate:	450 uL/min
Sample Injection:	6 uL
Solvent A:	95% water 5% acetonitrile 1 mM ammonium acetate
Solvent B:	95% acetonitrile 5% water 1 mM ammonium acetate
Chromatography Type:	Reversed phase

Chromatography ID:	CH003733
Chromatography Summary:	Positive C18
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
Column Temperature:	45
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
Sample Injection:	6 uL
Solvent A:	100% water; 0.1% formic acid
Solvent B:	100%acetonitrile; 0.1% formic acid
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004687
Analysis ID:	AN004944
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
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.
Ion Mode:	NEGATIVE

MS ID:	MS004688
Analysis ID:	AN004945
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
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.
Ion Mode:	POSITIVE