Body Fat As Determinant of Female Gonadal Dysfunction

Recruiting

• Conditions: Polycystic Ovary Syndrome; Hypothalamic Amenorrhea
• Interventions: Diagnostic Test: Anthropometric and physical examination; Diagnostic Test: Indirect calorimetry, accelerometer and seven-day dietary recall; Diagnostic Test: Biochemical, hormonal and metabolic phenotyping; Diagnostic Test: Sonographic studies; Diagnostic Test: 24-hour Ambulatory blood pressure monitoring; Procedure: Percutaneous biopsy; Diagnostic Test: Cardiovascular autonomic function studies; Diagnostic Test: Oral smear and feces specimen

• Registration Number: NCT03841981
• Lead Sponsor: Fundacion para la Investigacion Biomedica del Hospital Universitario Ramon y Cajal

Brief Summary

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.

Detailed Description

Not available

Study Timeline

• Study First Submitted: 2019-02-08
• Study First Submitted QC: 2019-02-12
• Study First Posted: 2019-02-15
• Study Start: 2020-01-31
• Status Verified: 2025-02
• Last Update Submitted: 2025-02-27
• Last Update Posted: 2025-03-03
• Primary Completion: 2025-06-30
• Study Completion: 2025-12-31

Recruitment & Eligibility

• Status: RECRUITING
• Sex: Female
• Target Recruitment: 50

Inclusion Criteria

Not provided

Exclusion Criteria

Not provided

Study & Design

• Study Type: OBSERVATIONAL
• Study Design: Not specified

Arm && Interventions

Group	Intervention	Description
V- Non-hyperandrogenic healthy women	Percutaneous biopsy	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
I- Hypothalamic amenorrhea	Anthropometric and physical examination	10 women with exercise-associated hypothalamic amenorrhea
III- Non-hyperandrogenic polycystic ovary syndrome	Indirect calorimetry, accelerometer and seven-day dietary recall	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
III- Non-hyperandrogenic polycystic ovary syndrome	Biochemical, hormonal and metabolic phenotyping	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
IV- Trained women without ovulatory dysfunction	Indirect calorimetry, accelerometer and seven-day dietary recall	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
IV- Trained women without ovulatory dysfunction	Cardiovascular autonomic function studies	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
I- Hypothalamic amenorrhea	24-hour Ambulatory blood pressure monitoring	10 women with exercise-associated hypothalamic amenorrhea
I- Hypothalamic amenorrhea	Cardiovascular autonomic function studies	10 women with exercise-associated hypothalamic amenorrhea
II- Hyperandrogenic polycystic ovary syndrome	Biochemical, hormonal and metabolic phenotyping	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
II- Hyperandrogenic polycystic ovary syndrome	Percutaneous biopsy	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
III- Non-hyperandrogenic polycystic ovary syndrome	Anthropometric and physical examination	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
III- Non-hyperandrogenic polycystic ovary syndrome	24-hour Ambulatory blood pressure monitoring	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
III- Non-hyperandrogenic polycystic ovary syndrome	Oral smear and feces specimen	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
IV- Trained women without ovulatory dysfunction	Biochemical, hormonal and metabolic phenotyping	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
IV- Trained women without ovulatory dysfunction	Sonographic studies	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
IV- Trained women without ovulatory dysfunction	Oral smear and feces specimen	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
II- Hyperandrogenic polycystic ovary syndrome	Anthropometric and physical examination	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
III- Non-hyperandrogenic polycystic ovary syndrome	Sonographic studies	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
III- Non-hyperandrogenic polycystic ovary syndrome	Percutaneous biopsy	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
IV- Trained women without ovulatory dysfunction	Anthropometric and physical examination	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
IV- Trained women without ovulatory dysfunction	24-hour Ambulatory blood pressure monitoring	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
V- Non-hyperandrogenic healthy women	Indirect calorimetry, accelerometer and seven-day dietary recall	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
V- Non-hyperandrogenic healthy women	Cardiovascular autonomic function studies	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
I- Hypothalamic amenorrhea	Indirect calorimetry, accelerometer and seven-day dietary recall	10 women with exercise-associated hypothalamic amenorrhea
I- Hypothalamic amenorrhea	Oral smear and feces specimen	10 women with exercise-associated hypothalamic amenorrhea
II- Hyperandrogenic polycystic ovary syndrome	24-hour Ambulatory blood pressure monitoring	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
II- Hyperandrogenic polycystic ovary syndrome	Cardiovascular autonomic function studies	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
III- Non-hyperandrogenic polycystic ovary syndrome	Cardiovascular autonomic function studies	5 lean women with non-hyperandrogenic polycystic ovary syndrome 5 women with weight excess and non-hyperandrogenic polycystic ovary syndrome
V- Non-hyperandrogenic healthy women	Biochemical, hormonal and metabolic phenotyping	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
V- Non-hyperandrogenic healthy women	24-hour Ambulatory blood pressure monitoring	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
I- Hypothalamic amenorrhea	Biochemical, hormonal and metabolic phenotyping	10 women with exercise-associated hypothalamic amenorrhea
I- Hypothalamic amenorrhea	Sonographic studies	10 women with exercise-associated hypothalamic amenorrhea
I- Hypothalamic amenorrhea	Percutaneous biopsy	10 women with exercise-associated hypothalamic amenorrhea
II- Hyperandrogenic polycystic ovary syndrome	Indirect calorimetry, accelerometer and seven-day dietary recall	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
II- Hyperandrogenic polycystic ovary syndrome	Sonographic studies	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
II- Hyperandrogenic polycystic ovary syndrome	Oral smear and feces specimen	5 lean women with hyperandrogenic polycystic ovary syndrome. 5 women with weight excess and hyperandrogenic polycystic ovary syndrome.
IV- Trained women without ovulatory dysfunction	Percutaneous biopsy	10 women who exercise as intensively as women with exercise-associated hypothalamic amenorrhea but with normal ovulatory cycles.
V- Non-hyperandrogenic healthy women	Anthropometric and physical examination	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
V- Non-hyperandrogenic healthy women	Sonographic studies	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
V- Non-hyperandrogenic healthy women	Oral smear and feces specimen	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

Primary Outcome Measures

Name	Time	Method
Adipokine and myokine signaling identification	Up to 5 years

Secondary Outcome Measures

Name	Time	Method
Circulating adipokine profile	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.).
Association between body mass index and sex steroids	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.
Association between body fat depots and sex steroids	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.
Appetite regulation hormonal profile	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.).
Association between percentage of fat mass with respect to total body weight and sex steroids	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.
Association between percentage of lean mass with respect to total body weight and sex steroids	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.
Association between body composition, sex steroids, and insulin resistance.	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.
Association between body composition, sex steroids, and lipids.	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.
Association between body composition, sex steroids, and office blood pressure.	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.
Association between body composition, sex steroids, and ambulatory blood pressure monitoring parameters.	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.
Association between body composition, sex steroids, and cardiovascular autonomic function tests.	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.
Association between body composition, sex steroids, and carotid intima-media thickness.	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.
Association between body composition, sex steroids, and oxidative stress.	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.
Association between body composition, sex steroids, and microbiome	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.

Trial Locations

Locations (1)

Endocrinology and Nutrition; 🇪🇸 Madrid, Spain