Body Fat as Determinant of Female Gonadal Dysfunction

Amount, Distribution and Dysfunction of Body Fat as Determinants of Female Gonadal Dysfunction: From Functional Hypothalamic Amenorrhea to the Polycystic Ovary Syndrome

Reproduction requires from women enough energy depots to warrant an adequate nutritional supply to the fetus. Hence, adipose tissue is able to communicate with female hypothalamic-pituitary-ovary axis. The hypothesis of the project is that abnormalities in the quantity (absolute and relative to lean body mass), distribution and/or function of adipose tissue are associated with functional forms of female gonadal dysfunction in predisposed women, in a spectrum of anomalies that go from hypothalamic amenorrhea to the polycystic ovary syndrome (PCOS). To challenge this hypothesis, the investigators will study 5 groups of 10 women each: women with exercise-associated hypothalamic amenorrhea, women without ovulatory dysfunction that exercise equally, non-hyperandrogenic patients with PCOS, hyperandrogenic patients with PCOS, and healthy control women comparable to those with PCOS. The aims of the study will be:

Primary objective: To identify novel signalling factors originating from adipose tissue and muscle using targeted and nontargeted evaluation of the proteome and of gene expression of superficial subcutaneous fat, deep subcutaneous fat (which mimics visceral adipose tissue) and skeletal muscle.

Secondary objectives:

  1. To study the serum adipokine profile - including those identified by the primary objective - and circulating gut hormones during fasting and after a glucose load in the 5 groups of women, and their associations with sexual hormones and body fat distribution.
  2. To study body composition and body fat distribution in these women and their relationships with:

2.1, Sex steroid profiles.

2.2. Classic cardiovascular risk factors: carbohydrate metabolism, lipid profiles and blood pressure.

2.3 Markers of low-grade chronic inflammation.

2.4. Oxidative stress markers.

2.5. Cardiovascular autonomic function.

2.6. Surrogate markers of subclinical atherosclerosis.

2.7. Circulating concentrations of endocrine disruptors.

2.8. Oral and gut microbiome.

The results will provide a better understanding of the mechanisms linking body energy depots with the female reproductive axis and, hopefully, the identification of potential biomarkers for the diagnosis and treatment of the disorders studied here.

Study Overview

Study Type

Observational

Enrollment (Anticipated)

50

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Locations

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

18 years to 40 years (Adult)

Accepts Healthy Volunteers

Yes

Genders Eligible for Study

Female

Sampling Method

Non-Probability Sample

Study Population

Patients with exercise-associated functional amenorrhea and polycystic ovary syndrome will be consecutively recruited from our Reproductive Endocrinolgy clinic. Those premenopausal adult women are referred to clinic because of symptoms of functional androgen excess such as hirsutism or menstrual dysfunction.

The control group (Groups IV and V) will include healthy female volunteers recruited - contemporarily with the recruitment of patients - from the hospital's staff and overweight or obese women seeking medical attention in our department.

Description

Inclusion Criteria

Group I

  • Body mass index between 18.5 and 25.0 kg/m2.
  • Group 1 ovulatory dysfunction [World Health Organization (WHO) classification].
  • Normal/low gonadotrophin levels [follicle-stimulating hormone (FSH) and luteinizing (LH) < 10 IU/l] and low estradiol (< 50 pg/ml).
  • Moderate-vigorous intensity physical activity (> 5 hours per week) plus low energy availability (< 30 kcal/per kg of lean mass).
  • Exclusion of secondary etiologies
  • Informed consent signed.

Group II:

  • Polycystic ovary syndrome phenotype I, II and III [National Institute of Health (NIH)-2012] with hyperandrogenemia (http://prevention.nih.gov/workshops/2012/resources.aspx).
  • Body mass index between 18.5 and 40.0 kg/m2.
  • Informed consent signed.

Group III:

  • Polycystic ovary syndrome phenotype IV (NIH-2012) (http://prevention.nih.gov/workshops/2012/resources.aspx).
  • Body mass index between 18.5 and 40.0 kg/m2.
  • Informed consent signed.

Group IV:

  • Body mass index between 18.5 and 25.0 kg/m2.
  • Regular menses.
  • Normal gonadotropins and estradiol levels at follicular phase.
  • Moderate-vigorous intensity physical activity (> 5 hours per week) with normal energy availability (> 30 kcal/per kg of lean mass).
  • Informed consent signed.

Group V:

  • No signs or symptoms of hyperandrogenism.
  • No exercise or mild intensity physical activity.
  • Regular menses.
  • Body mass index between 18.5 and 40.0 kg/m2.
  • Informed consent signed.

Exclusion Criteria (Groups I-V)

  • Oral drugs interfering with ovulation (glucocorticoids, antipsychotics, antidepressants, contraceptives, sex steroids and/or opioids) for the previous 6 months to study inclusion.
  • Current pregnancy or lactation, or during the previous 6 months to study inclusion.
  • Asherman's syndrome or outflow tract disorders.
  • Current smoking or alcohol intake > 40 g per day.
  • Previous diagnosis of glucose intolerance, hypertension, dyslipidemia, known heart or lung diseases, kidney disease, liver disease, celiac disease or any other malabsorptive condition, chronic inflammatory disease or malignancy.

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Observational Models: Case-Control
  • Time Perspectives: Cross-Sectional

Cohorts and Interventions

Group / Cohort
Intervention / Treatment
I- Hypothalamic amenorrhea
10 women with exercise-associated hypothalamic amenorrhea
  • Weight and height.
  • Waist-to-hip ratio.
  • Body composition: Bioelectrical impedance and [Dual energy X-ray absorptiometry (DEXA)].
Energy availability assessment.
  • Lipid profile.
  • Oral glucose tolerance test: plasma glucose and insulin, insulin sensitivity indices, gastrointestinal hormones, adipokines, oxidative stress markers.
  • Sex steroid profile.
  • Hypothalamic-pituitary-adrenal axis study.
  • Ferrokinetic study.
  • Subclinical chronic inflammatory markers.
  • Polycystic ovarian morphology.
  • Carotid intima-media thickness.
  • Eco-FAT: Ultrasound measurements of adipose tissue depots including sc, preperitoneal, intraperitoneal (ip), mesenteric, and perirenal fat thickness.
A&D TM2430EX oscillometric devices (A&D Company Limited, Tokyo, Japan).
Subcutaneous fat tissue and muscle tissue for proteomics an gene expression studies.
Parasympathetic and sympathetic responses to deep breathing, Valsalva's maneuver and orthostatism.
Microbiome studies.
II- Hyperandrogenic polycystic ovary syndrome
5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
  • Weight and height.
  • Waist-to-hip ratio.
  • Body composition: Bioelectrical impedance and [Dual energy X-ray absorptiometry (DEXA)].
Energy availability assessment.
  • Lipid profile.
  • Oral glucose tolerance test: plasma glucose and insulin, insulin sensitivity indices, gastrointestinal hormones, adipokines, oxidative stress markers.
  • Sex steroid profile.
  • Hypothalamic-pituitary-adrenal axis study.
  • Ferrokinetic study.
  • Subclinical chronic inflammatory markers.
  • Polycystic ovarian morphology.
  • Carotid intima-media thickness.
  • Eco-FAT: Ultrasound measurements of adipose tissue depots including sc, preperitoneal, intraperitoneal (ip), mesenteric, and perirenal fat thickness.
A&D TM2430EX oscillometric devices (A&D Company Limited, Tokyo, Japan).
Subcutaneous fat tissue and muscle tissue for proteomics an gene expression studies.
Parasympathetic and sympathetic responses to deep breathing, Valsalva's maneuver and orthostatism.
Microbiome studies.
III- Non-hyperandrogenic polycystic ovary syndrome
5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
  • Weight and height.
  • Waist-to-hip ratio.
  • Body composition: Bioelectrical impedance and [Dual energy X-ray absorptiometry (DEXA)].
Energy availability assessment.
  • Lipid profile.
  • Oral glucose tolerance test: plasma glucose and insulin, insulin sensitivity indices, gastrointestinal hormones, adipokines, oxidative stress markers.
  • Sex steroid profile.
  • Hypothalamic-pituitary-adrenal axis study.
  • Ferrokinetic study.
  • Subclinical chronic inflammatory markers.
  • Polycystic ovarian morphology.
  • Carotid intima-media thickness.
  • Eco-FAT: Ultrasound measurements of adipose tissue depots including sc, preperitoneal, intraperitoneal (ip), mesenteric, and perirenal fat thickness.
A&D TM2430EX oscillometric devices (A&D Company Limited, Tokyo, Japan).
Subcutaneous fat tissue and muscle tissue for proteomics an gene expression studies.
Parasympathetic and sympathetic responses to deep breathing, Valsalva's maneuver and orthostatism.
Microbiome studies.
IV- Trained women without ovulatory dysfunction
10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
  • Weight and height.
  • Waist-to-hip ratio.
  • Body composition: Bioelectrical impedance and [Dual energy X-ray absorptiometry (DEXA)].
Energy availability assessment.
  • Lipid profile.
  • Oral glucose tolerance test: plasma glucose and insulin, insulin sensitivity indices, gastrointestinal hormones, adipokines, oxidative stress markers.
  • Sex steroid profile.
  • Hypothalamic-pituitary-adrenal axis study.
  • Ferrokinetic study.
  • Subclinical chronic inflammatory markers.
  • Polycystic ovarian morphology.
  • Carotid intima-media thickness.
  • Eco-FAT: Ultrasound measurements of adipose tissue depots including sc, preperitoneal, intraperitoneal (ip), mesenteric, and perirenal fat thickness.
A&D TM2430EX oscillometric devices (A&D Company Limited, Tokyo, Japan).
Subcutaneous fat tissue and muscle tissue for proteomics an gene expression studies.
Parasympathetic and sympathetic responses to deep breathing, Valsalva's maneuver and orthostatism.
Microbiome studies.
V- Non-hyperandrogenic healthy women
10 women matched by age and body mass index with women with polycystic ovary syndrome who do not perform physical activity on a regular basis
  • Weight and height.
  • Waist-to-hip ratio.
  • Body composition: Bioelectrical impedance and [Dual energy X-ray absorptiometry (DEXA)].
Energy availability assessment.
  • Lipid profile.
  • Oral glucose tolerance test: plasma glucose and insulin, insulin sensitivity indices, gastrointestinal hormones, adipokines, oxidative stress markers.
  • Sex steroid profile.
  • Hypothalamic-pituitary-adrenal axis study.
  • Ferrokinetic study.
  • Subclinical chronic inflammatory markers.
  • Polycystic ovarian morphology.
  • Carotid intima-media thickness.
  • Eco-FAT: Ultrasound measurements of adipose tissue depots including sc, preperitoneal, intraperitoneal (ip), mesenteric, and perirenal fat thickness.
A&D TM2430EX oscillometric devices (A&D Company Limited, Tokyo, Japan).
Subcutaneous fat tissue and muscle tissue for proteomics an gene expression studies.
Parasympathetic and sympathetic responses to deep breathing, Valsalva's maneuver and orthostatism.
Microbiome studies.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Time Frame
Adipokine and myokine signaling identification
Time Frame: Up to 5 years
Up to 5 years

Secondary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Circulating adipokine profile
Time Frame: Up to 5 years
At fasting and after an oral glucose challenge: Circulating concentrations of Leptin, Adiponectin, Chemerin, Lipocalin-2, Adipsin, Plasminogen Activator Inhibitor (PAI)-1, Monocyte Chemoattractant Protein (MCP)-1, and Soluble Leptin Receptor by multianalyte profiling on the Luminex Magpix system (Luminex Technologies, Austin, USA.).
Up to 5 years
Appetite regulation hormonal profile
Time Frame: Up to 5 years
At fasting and after an oral glucose challenge: Circulating concentrations of Amylin, C-Peptide, Ghrelin, Gastric Inhibitory Peptide (GIP), Glucagon-Like Peptide (GLP)-1, Glucagon, IL-6, Insulin, Pancreatic Polypeptide (PP), Peptide YY, Tumor Necrosis Factor (TNF)-α by multianalyte profiling on the Luminex Magpix system (Luminex Technologies, Austin, USA.).
Up to 5 years
Association between body mass index and sex steroids
Time Frame: Up to 5 years
Body mass index in in kg/m^2. Sex steroids (including circulating total testosterone, estradiol, androstenedione, dehydroepiandrosterone-sulphate and estrone) measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). Free testosterone will be calculated from total testosterone and sex hormone binding globulin levels.
Up to 5 years
Association between percentage of fat mass with respect to total body weight and sex steroids
Time Frame: Up to 5 years
Fat mass% by bioelectric impedanciometry and DEXA. Sex steroids (including circulating total testosterone, estradiol, androstenedione, dehydroepiandrosterone-sulphate and estrone) measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). Free testosterone will be calculated from total testosterone and sex hormone binding globulin levels.
Up to 5 years
Association between percentage of lean mass with respect to total body weight and sex steroids
Time Frame: Up to 5 years
Lean mass% by bioelectric impedanciometry and DEXA. Sex steroids (including circulating total testosterone, estradiol, androstenedione, dehydroepiandrosterone-sulphate and estrone) measured by liquid chromatography tandem mass spectrometry (LC-MS/MS). Free testosterone will be calculated from total testosterone and sex hormone binding globulin levels.
Up to 5 years
Association between body fat depots and sex steroids
Time Frame: Up to 5 years
Adipose tissue depots will be estimated using a Toshiba Nemio XG SSA-580A Diagnostic Ultrasound System. Minimum sc and preperitoneal fat thicknesses will be measured at the level of the xyphoid process. Maximum sc fat thickness will be measured at the level of the umbilicus. Intraperitoneal fat thickness will be measured placing a probe transversally in the midline of abdomen, 2 cm above the umbilicus. Three measures of ip fat thickness will be obtained: the distance from the fascia of rectus abdominis muscle to the vertebral column, the distance from the peritoneum to the vertebral column, and the distance from the linea alba to the vertebral column. Perirenal fat thickness will be estimated as the distance from the perirenal fascia to the renal surface. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and insulin resistance.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Fasting glucose and insulin levels will be used for calculating the homeostasis model assessment of insulin resistance (HOMA-IR), and the composite insulin sensitivity index will be estimated from the glucose and insulin concentrations measured during the oral glucose tolerance test. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and lipids.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Circulating HDL-cholesterol and phospholipid levels will be measured by enzymatic methods after precipitation of plasma with phosphotungstic acid and Mg2+. Total cholesterol and triglyceride levels will be determined by enzymatic methods. LDL-cholesterol concentrations will be estimated by Friedewald's equation. Circulating apolipoprotein (Apo) AI, Apo B100, and lipoprotein (a) levels will be determined by kinetic immunonephelometry. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and office blood pressure.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Office blood pressure will be determined as the mean of three manual sphygmomanometer readings in the sitting position. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and ambulatory blood pressure monitoring parameters.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Twenty-four-hour ambulatory blood pressure monitoring will be performed using an A&D TM2430EX oscillometric device (A&D Co., Ltd., Tokyo, Japan). The cuff (12 × 22 cm for lean participants, 14 × 30 cm for overweight or obese participants) will placed on the nondominant arm in every woman. The period from 0700 to 2300 h will be considered daytime, and from 2300 until 0700 h the next day will be considered nighttime, reflecting the usual sleeping habits of Spaniards. Systolic, diastolic, and mean blood pressure as well as heart rate will be measured every 20 min during daytime and every 30 min during nighttime. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and cardiovascular autonomic function tests.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Cardiovascular autonomic function will be assessed by the blood pressure and heart rate responses to active standing, and Ewing and Clarke's tests. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and carotid intima-media thickness.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Imaging will be conducted using a high-resolution 7.5-MHz phased-array transducer by the same trained operator in all the participants. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and oxidative stress.
Time Frame: Up to 5 years
Fat mass, lean mass and body fat depots will be measured as previously described. Oxidative stress profile will be measured by enzymatic assays: Plasma thiobarbituric acid reactive substances, total antioxidant capacity, nitrotyrosine, protein carbonyl groups and erythrocyte glutathione peroxidase levels. Sex steroids will be measured as previously described.
Up to 5 years
Association between body composition, sex steroids, and microbiome
Time Frame: Up to 5 years
Participants will be instructed to collect fecal and salivary samples. DNA samples wil be used for massive sequencing of 16S ribosomal DNA (rDNA) amplicons in a MiSeq platform (Illumina). The bacterial diversity will be estimated by using Shannon, Chao 1, Jaccard, and Sorensen indexes with their SDs. Taxonomic affiliations will be assigned by using the RDP_classifier from the Ribosomal Database Project (RDP), and readings with RDP score value <0.8 will be assigned to the upper taxonomic rank, leaving the last rank as unidentified. Sex steroids will be measured as previously described.
Up to 5 years

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Investigators

  • Principal Investigator: Héctor F Escobar-Morreale, PhD, MD, Diabetes, Obesity and Human Reproduction Research Group (CIBERDEM), Department of Endocrinology and Nutrition, Hospital Universitario Ramón y Cajal, Universidad de Alcalá, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain
  • Principal Investigator: Manuel Luque-Ramírez, PhD, MD, Diabetes, Obesity and Human Reproduction Research Group (CIBERDEM), Department of Endocrinology and Nutrition, Hospital Universitario Ramón y Cajal, Universidad de Alcalá, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain

Publications and helpful links

The person responsible for entering information about the study voluntarily provides these publications. These may be about anything related to the study.

General Publications

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

January 31, 2020

Primary Completion (Anticipated)

December 31, 2023

Study Completion (Anticipated)

December 31, 2023

Study Registration Dates

First Submitted

February 8, 2019

First Submitted That Met QC Criteria

February 12, 2019

First Posted (Actual)

February 15, 2019

Study Record Updates

Last Update Posted (Actual)

March 1, 2023

Last Update Submitted That Met QC Criteria

February 28, 2023

Last Verified

February 1, 2023

More Information

Terms related to this study

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

YES

IPD Plan Description

The investigators will deposit any datasets and supplemental materials to a generalist repository. The investigators will refer to the deposited material as an item in the bibliography of any manuscript and number the reference appropriately within the text.

IPD Sharing Time Frame

Data will become available at the time of the first manuscript submission.

IPD Sharing Access Criteria

Public access

IPD Sharing Supporting Information Type

  • STUDY_PROTOCOL
  • SAP
  • ICF
  • ANALYTIC_CODE
  • CSR

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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