Development of Biomarkers for Monitoring Menopause in Women

This cross-sectional study aims to develop and validate mitochondrial biomarkers for monitoring menopause in women. The study will involve 100 participants divided into three groups: reproductive period, perimenopausal transition period, and postmenopausal period. Peripheral blood samples will be collected, and mitochondrial quality and quantity will be assessed using both qualitative and quantitative analyses. The study will identify potential biomarkers through metabolomics and validate them for monitoring the perimenopausal transition, providing a novel approach for women's health management during menopause.

Study Overview

• Status: Recruiting
• Conditions: Perimenopausal Biomarkers; Mitochondrial Biomarkers
• Intervention / Treatment: Other: Blood Sample Collection

Detailed Description

Background： Gender equality and equitable health have emerged as international trends in recent years. Despite existing solutions for women's health issues, many needs remain unmet. According to projections, the global menopausal population is expected to reach 1.2 billion by 2030, surpassing the prevalence of the most common chronic diseases. Presently, solutions exist for common menopausal symptoms such as hot flashes, night sweats, fatigue, irritability, and urinary incontinence. However, unresolved challenges encompass psychological changes and monitoring systems. Therefore, this project aims to develop biomarkers for menopause in women as a future monitoring system.

Most women enter menopause between the ages of 48 and 52. During this period, ovarian follicles cease activity, leading to the one-year period following the appearance of significant menstrual irregularities, known as the menopausal transition or perimenopause (1). As ovarian function gradually declines and estrogen secretion diminishes, various systemic changes occur, resulting in several menopause-related conditions such as osteoporosis, cardiovascular diseases, and urinary system issues. The decline in estrogen triggers a negative feedback loop, causing an increase in follicle-stimulating hormone (FSH) secretion and a reduction in E2 (17β-estradiol) production (2). Currently, menopausal hormone therapy (MHT) using estrogen can alleviate menopausal symptoms and restore overall E2 levels but doesn't lower FSH to premenopausal levels (3-4). However, a precise biomarker for monitoring menopause has not yet been established. Under estrogen deficiency, mitochondrial morphology and function may be compromised, while estrogen presence relates to reduced oxidative reactions, enhanced respiratory function, and stable membrane properties (5). This project aims to provide a comprehensive view through metabolomics to identify biomarkers within mitochondria that can be used for menopause monitoring.

Study Design： This study is a cross-sectional study. A total of 100 women will be included and grouped into three categories: 25 cases of reproductive period, 50 cases of perimenopausal transition period, and 25 cases of postmenopausal period. The peripheral blood will be collected. Following mitochondrial extraction, both qualitative and quantitative analyses are conducted. High-confidence protein markers are identified as candidate molecules using combined data from RNA-seq and LC/MS/MS. These candidate markers will undergo further validation testing.

Methods： After Ficoll separation, peripheral blood will be divided into mononuclear cells, plasma, and red blood cells. Further extraction of mitochondria will be carried out from both mononuclear cells and plasma. Quantitative experiments involve hippocampal measurements of metabolic energy and JC-1 assessment of mitochondrial health, while mitochondrial DNA copy number will be evaluated using digital PCR. Qualitative experimental analysis employs Western blotting to assess markers such as cytochrome c and HSP60 for mitochondria and PCNA for nuclear proteins. Candidate biomarker validation experiments use combined data from RNA-seq and LC/MS/MS to identify high-confidence protein markers as candidates, which are further validated for feasibility using the ELISA method.

Effect：

1. Understand the changes in mitochondrial quality and quantity during the perimenopausal transition in women.
2. Develop monitoring indicators for the perimenopausal transition in women.

• Study Type: Observational
• Enrollment (Estimated): 100

Contacts and Locations

• Study Contact: Name: Shu-Yu Liu; Phone Number: +886910893512; Email: syliu0123@outlook.com

Study Locations

• Taiwan
  • New Taipei City, Taiwan, 24352
    • Recruiting
    • Fu Jen Catholic University Hospital, Fu Jen Catholic University
    • Contact: Shu-Yu Liu; Phone Number: +886910893512; Email: syliu0123@outlook.com

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: Yes

• Sampling Method: Non-Probability Sample

Study Population

Study Population Description: The study population consists of adult women aged over 20 years old, categorized into three groups:

reproductive period (ages 20-45, regular menstrual cycles) perimenopausal transition period (ages 45-50, experiencing menstrual irregularities) postmenopausal period (ages over 50, no menstrual period for at least one year).

All participants must be in general good health, without major chronic diseases, infection, or pregnancy.

Description

Inclusion Criteria:

• Biological female, age order than 20 years old.

Exclusion Criteria:

• Person with cancer.
• Person with any known acute or chronic infection.
• Person with known chronic illness under follow up or treatment.
• Pregnancy, one year withing delivery, under breast feeding, or three months within breast feeding.
• Under any female horemone therapy.

Study Plan

How is the study designed?

Number of groups / cohorts

3

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Reproductive Period; Women aged 20-40 years with regular menstrual cycles, representing the reproductive stage.	Other: Blood Sample Collection; Collection of peripheral blood samples for mitochondrial analysis, including the extraction of mononuclear cells and plasma for further assessment of mitochondrial quality and quantity.
Perimenopausal Transition Period; Women aged 40-55 years have menstrual cycle within one year but experiencing menstrual irregularities, representing the transition into menopause.	Other: Blood Sample Collection; Collection of peripheral blood samples for mitochondrial analysis, including the extraction of mononuclear cells and plasma for further assessment of mitochondrial quality and quantity.
Postmenopausal Period; Women aged 50 years and older who have not had a menstrual period for at least one year, representing the postmenopausal stage.	Other: Blood Sample Collection; Collection of peripheral blood samples for mitochondrial analysis, including the extraction of mononuclear cells and plasma for further assessment of mitochondrial quality and quantity.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Mitochondrial Quantitative Analysis	This outcome measure involves evaluating mitochondrial energy metabolism, including ATP content, oxygen consumption rate (OCR), and extracellular acidification rate (ECAR) using a hippocampal metabolic analyzer. Additionally, mitochondrial membrane potential will be assessed using flow cytometry to analyze the distribution of JC-1, indicating mitochondrial health. Digital PCR will be employed to measure mitochondrial DNA copy numbers, determining mitochondrial mass and comparing differences between groups.	12 months from study start
Mitochondrial Qualitative Analysis	This measure assesses mitochondrial protein markers using Western blot analysis. PCNA will be used as a nuclear protein marker, while cytochrome c and HSP60 will be used as mitochondrial protein markers to evaluate mitochondrial quality	12 months from study start
Validation of Candidate Biomarkers	This outcome measure focuses on the validation of high-confidence protein markers identified through RNA-Seq and LC-MS/MS data. The validation will be conducted using customized protein databases and further experiments to confirm the feasibility of these candidate biomarkers for monitoring perimenopausal transition.	12 months from study start

Collaborators and Investigators

• Sponsor: Fu Jen Catholic University Hospital

Publications and helpful links

General Publications

• Davis SR, Lambrinoudaki I, Lumsden M, Mishra GD, Pal L, Rees M, Santoro N, Simoncini T. Menopause. Nat Rev Dis Primers. 2015 Apr 23;1:15004. doi: 10.1038/nrdp.2015.4.
• Pellegrino A, Tiidus PM, Vandenboom R. Mechanisms of Estrogen Influence on Skeletal Muscle: Mass, Regeneration, and Mitochondrial Function. Sports Med. 2022 Dec;52(12):2853-2869. doi: 10.1007/s40279-022-01733-9. Epub 2022 Jul 30.
• Karppinen JE, Tormakangas T, Kujala UM, Sipila S, Laukkanen J, Aukee P, Kovanen V, Laakkonen EK. Menopause modulates the circulating metabolome: evidence from a prospective cohort study. Eur J Prev Cardiol. 2022 Aug 5;29(10):1448-1459. doi: 10.1093/eurjpc/zwac060.
• Powers MS, Schenkel L, Darley PE, Good WR, Balestra JC, Place VA. Pharmacokinetics and pharmacodynamics of transdermal dosage forms of 17 beta-estradiol: comparison with conventional oral estrogens used for hormone replacement. Am J Obstet Gynecol. 1985 Aug 15;152(8):1099-106. doi: 10.1016/0002-9378(85)90569-1.

Study record dates

Study Major Dates

• Study Start (Actual): July 30, 2024
• Primary Completion (Estimated): February 1, 2026
• Study Completion (Estimated): August 1, 2026

Study Registration Dates

• First Submitted: February 10, 2025
• First Submitted That Met QC Criteria: February 24, 2025
• First Posted (Actual): February 28, 2025

Study Record Updates

• Last Update Posted (Actual): May 25, 2025
• Last Update Submitted That Met QC Criteria: May 20, 2025
• Last Verified: February 1, 2025

More Information

Terms related to this study

• Keywords: Biomarker; Women's Health; Mitochondria; Perimenopausal Transition
• Other Study ID Numbers: FJUH112331

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No