Summary of Study ST003044

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	ST003044
Study Title	A High-Fat Eucaloric Diet Induces Reprometabolic Syndrome of Obesity in Normal Weight Women - lipidomics
Study Summary	Lipidomics analysis was performed on plasma from human female subjects before, during, and after a high fat diet.
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	2023-12-15
Raw Data Available	Yes
Raw Data File Type(s)	raw(Thermo)
Analysis Type Detail	LC-MS
Release Date	2024-02-05
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:	SU003159
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
SA330443	DS1-28-49	On-diet
SA330444	DS1-28-02	On-diet
SA330445	DS1-28-43	On-diet
SA330446	DS1-28-41	On-diet
SA330447	DS1-28-52	On-diet
SA330448	DS1-28-56	On-diet
SA330449	DS1-28-54	On-diet
SA330450	DS1-28-38	On-diet
SA330451	DS1-28-46	On-diet
SA330452	DS1-28-11	On-diet
SA330453	DS1-28-35	On-diet
SA330454	DS1-28-05	On-diet
SA330455	DS1-28-14	On-diet
SA330456	DS1-28-08	On-diet
SA330457	DS1-28-17	On-diet
SA330458	DS1-28-26	On-diet
SA330459	DS1-28-29	On-diet
SA330460	DS1-28-20	On-diet
SA330461	DS1-28-36	Post-diet
SA330462	DS1-28-30	Post-diet
SA330463	DS1-28-39	Post-diet
SA330464	DS1-28-50	Post-diet
SA330465	DS1-28-47	Post-diet
SA330466	DS1-28-44	Post-diet
SA330467	DS1-28-27	Post-diet
SA330468	DS1-28-03	Post-diet
SA330469	DS1-28-21	Post-diet
SA330470	DS1-28-06	Post-diet
SA330471	DS1-28-09	Post-diet
SA330472	DS1-28-18	Post-diet
SA330473	DS1-28-15	Post-diet
SA330474	DS1-28-12	Post-diet
SA330475	DS1-28-45	Pre-diet
SA330476	DS1-28-42	Pre-diet
SA330477	DS1-28-48	Pre-diet
SA330478	DS1-28-55	Pre-diet
SA330479	DS1-28-37	Pre-diet
SA330480	DS1-28-53	Pre-diet
SA330481	DS1-28-51	Pre-diet
SA330482	DS1-28-40	Pre-diet
SA330483	DS1-28-19	Pre-diet
SA330484	DS1-28-07	Pre-diet
SA330485	DS1-28-04	Pre-diet
SA330486	DS1-28-01	Pre-diet
SA330487	DS1-28-10	Pre-diet
SA330488	DS1-28-13	Pre-diet
SA330489	DS1-28-28	Pre-diet
SA330490	DS1-28-25	Pre-diet
SA330491	DS1-28-16	Pre-diet
SA330492	DS1-28-34	Pre-diet

Showing results 1 to 50 of 50

Showing results 1 to 50 of 50

Collection:

Collection ID:	CO003152
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:	TR003168
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 0-240 minutes) and concluded with a physiologic bolus of 75 ng/kg GnRH to assess pituitary responsiveness (Time 240-360 minutes). Frequent sampling studies for gonadotropin determination were all performed in the early follicular phase of the menstrual cycle (cycle days 2-5). Participants were also seen weekly during the high-fat diet cycle and had a blood draw to assess compliance. High-fat diet protocol. Participants completed a food preference screening survey and a 3-day diet diary, which estimated their daily caloric intake and approximate proportion of daily calories derived from fat. Participants were provided with a customized eucaloric high-fat diet comprised of approximately 48% calories from fat, 33% calories from carbohydrates and 19% calories from protein. Food was portioned into individual meals with snacks and boxed into 3-4 days’ worth at a time. Participants picked up food directly from the Clinical and Translational Research Center; when this was not possible, study staff delivered food to the participant. Urinary ketones were measured at each frequent blood sampling session (Siemens Healthineers, Malvern, PA). During administration of the high-fat diet, blood samples were drawn weekly for metabolomic analysis to determine compliance with the protocol.

Treatment ID:

TR003168

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 0-240 minutes) and concluded with a physiologic bolus of 75 ng/kg GnRH to assess pituitary responsiveness (Time 240-360 minutes). Frequent sampling studies for gonadotropin determination were all performed in the early follicular phase of the menstrual cycle (cycle days 2-5). Participants were also seen weekly during the high-fat diet cycle and had a blood draw to assess compliance. High-fat diet protocol. Participants completed a food preference screening survey and a 3-day diet diary, which estimated their daily caloric intake and approximate proportion of daily calories derived from fat. Participants were provided with a customized eucaloric high-fat diet comprised of approximately 48% calories from fat, 33% calories from carbohydrates and 19% calories from protein. Food was portioned into individual meals with snacks and boxed into 3-4 days’ worth at a time. Participants picked up food directly from the Clinical and Translational Research Center; when this was not possible, study staff delivered food to the participant. Urinary ketones were measured at each frequent blood sampling session (Siemens Healthineers, Malvern, PA). During administration of the high-fat diet, blood samples were drawn weekly for metabolomic analysis to determine compliance with the protocol.

Sample Preparation:

Sampleprep ID:	SP003165
Sampleprep Summary:	Lipids from 10 uL of plasma were extracted with 90 uL of cold methanol. Samples were briefly vortexed at room temp then incubated at -20 C for 30 min. Supernatants were clarified by centrifugation (10 min, 10,000 g, 4 degrees C) and transferred to autosampler vials.
Processing Storage Conditions:	4℃
Extract Storage:	-80℃

Combined analysis:

Analysis ID	AN004993	AN004994
Analysis type	MS	MS
Chromatography type	Reversed phase	Reversed phase
Chromatography system	Thermo Vanquish	Thermo Vanquish
Column	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)	Phenomenex Kinetex C18 (30 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:	CH003772
Chromatography Summary:	Negative C18
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)
Column Temperature:	45
Flow Gradient:	0-3 min 10-95% B at 0.3 mL/min, 3-4.2 min hold at 95% B at 0.3 mL/min, 4.2-4.3 min drop to 10% B at 0.45 mL/min, 4.3-4.9 min hold at 10%B while lowering flow rate to 0.4 mL/min, 4.9-5 min hold at 10%B while lowering flow rate to 0.3 mL/min.
Flow Rate:	0.3-0.4 ml/min
Sample Injection:	10 uL
Solvent A:	75% water/25% acetonitrile; 5 mM ammonium acetate
Solvent B:	90% isopropanol/10% acetonitrile; 5 mM ammonium acetate
Chromatography Type:	Reversed phase

Chromatography ID:	CH003773
Chromatography Summary:	Positive C18
Instrument Name:	Thermo Vanquish
Column Name:	Phenomenex Kinetex C18 (30 x 2.1mm,1.7um)
Column Temperature:	45
Flow Gradient:	0-3 min 30-100% B at 0.3 mL/min, 3-4.2 min hold at 100% B at 0.3 mL/min, 4.2-4.3 min 100-30% B at 0.4 mL/min, 4.3-4.9 min hold at 30%B and 0.4 mL/min, 4.9-5 min hold at 30%B while lowering flow rate to 0.3 mL/min
Flow Rate:	0.3-0.4 ml/min
Sample Injection:	10 uL
Solvent A:	75% water/25% acetonitrile; 5 mM ammonium acetate
Solvent B:	90% isopropanol/10% acetonitrile; 5 mM ammonium acetate
Chromatography Type:	Reversed phase

MS:

MS ID:	MS004733
Analysis ID:	AN004993
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control.
Ion Mode:	NEGATIVE

MS ID:	MS004734
Analysis ID:	AN004994
Instrument Name:	Thermo Q Exactive Orbitrap
Instrument Type:	Orbitrap
MS Type:	ESI
MS Comments:	The Q Exactive was run independently in positive and negative ion mode, scanning using full MS from 125-1500 m/z at 70,000 resolution and top 10 data-dependent MS2 at 17,500 resolution. Electrospray ionization was achieved with 45 Arb sheath gas, 25 Arb auxiliary gas, and 4 kV spray voltage. Calibration was performed prior to the run using the PierceTM Positive and Negative Ion Calibration Solutions (Thermo Fisher Scientific). Run order of samples was randomized and technical replicates were injected after every 4 samples to assess quality control.
Ion Mode:	POSITIVE