LINE-1 and Alu Methylation Levels Among Middle Aged Women With Low Cardiovascular Risk Profile in Respect of Menopausal Hot Flashes

NCT05892211 | Suspended

• 1 other identifier: E-83045809-604.01.02-3916
• Study Type: observational
• Target: 30
• Locations: 1 country
• Sites: 1

Brief Summary

Vasomotor symptoms are the most common symptoms seen during climacterium. The hypoestrogenic state causes dysfunction of hypothalamic preoptic area, a thermoregulatory center. The sympathetic overactivation during the hot flashes is associated with awakening during sleep and have a negative impact on cardiac indexes and vascular reactivity. Therefore, hot flashes are accepted as subclinical cardiovascular risk factor. The association between the severity of the hot flashes and cardiovascular risk may have an epigenetic background. Recently, methylation changes of DNA was found to be associated with clinical and subclinical cardiovascular disease risk (atherosclerosis and hypertension etc.). A transposable element in the DNA, Long interspersed nuclear elements (LINE-1), was found to be hypomethylated in cases with ischemic heart disease and stroke. Therefore, the expression of repeating elements in the DNA (LINE-1 and ALU) may be considered as a mediator in the ischemic heart disease. Until now, menopausal age, vasomotor symptoms and epigenetic and biological aging have been evaluated. However, the epigenetic impact of severe vasomotor symptoms in postmenopausal women with low cardiovascular disease risk profile has not been evaluated. In this study, we aimed to evaluate the epigenetic basis of cardiovascular disease risk for women with vasomotor symptoms which disturb sleep by assessing the methylation levels of ALU and LINE-1.

Conditions

Hot Flashes; Sleep; Menopause; Cardiovascular Diseases; Epigenetics; DNA Methylation

Keywords

DNA methylation; Hot Flashes; Sleep; Vasomotor symptoms; Menopause; Cardiovascular Diseases; epigenetics

Outcome Measures

Primary Outcomes (6)

• Methylation levels of ALU and LINE-1

  Methylation levels of ALU and LINE-1 in women with vasomotor symptoms

  1 day

• Polysomnographic findings

  Total sleep time in women with vasomotor symptoms

  1 night

• Number of objective hot flashes per night

  Number of objective hot flashes per night measured by sternal skin conductance in women with vasomotor symptoms

  1 night

• Number of nighttime awakenings per night

  Number of nighttime awakenings per night associated with hot flash episodes

  1 night

• Polysomnographic findings

  Sleep efficiency in women with vasomotor symptoms

  1 night

• Polysomnographic findings

  Sleep stages in women with vasomotor symptoms

  1 night

Secondary Outcomes (2)

• Subjective sleep findings (Pittsburgh Sleep Quality Index (PSQI))

  1 night

• Menopause-Specific Quality of Life Questionnaire (MENQOL) scores and its association with hot flashes per night

  1 day

Study Arms (2)

Group 1 (vasomotor symptoms present)

Participants who have vasomotor symptoms. Sleep is recorded by polysomnography for a night. Hot flashes during sleep are recorded by two electrodes measuring the skin conductance near sternum. Any hot flashes reported subjectively by the patient during the recording is noted. Their blood samples are obtained.

Diagnostic Test: Polysomnography; Diagnostic Test: Skin conductance; Genetic: ALU and LINE-1 DNA methylation analysis; Diagnostic Test: The Menopause-Specific Quality of Life Questionnaire (MENQOL); Diagnostic Test: Pittsburgh Sleep Quality Index (PSQI)

Group 2 (vasomotor symptoms absent)

Participants who don't have vasomotor symptoms. Sleep is recorded by polysomnography for a night. Hot flashes during sleep are recorded by two electrodes measuring the skin conductance near sternum. Any hot flashes reported subjectively by the patient during the recording is noted. Their blood samples are obtained.

Diagnostic Test: Polysomnography; Diagnostic Test: Skin conductance; Genetic: ALU and LINE-1 DNA methylation analysis; Diagnostic Test: The Menopause-Specific Quality of Life Questionnaire (MENQOL); Diagnostic Test: Pittsburgh Sleep Quality Index (PSQI)

Interventions

Polysomnography DIAGNOSTIC_TEST

Polysomnography is performed by 3-channel electroencephalography ((EEG; F4-M1, C4-M1, O2-M1), 2-channel electrooculography (EOG), chin electromyography (EMG), surface EMG recording from tibialis anterior muscles of the right and left leg, body position, oronasal thermal airflow sensor, nasal pressure sensor, thoracic and abdominal respiratory belts for assesing the respiratory effort, electrocardiography (ECG), pulse, recording of respiratory sounds, oxygen saturation and synchronous video recording. The sleep stages are scored by the current criteria of American Academy of Sleep Medicine.

Group 1 (vasomotor symptoms present); Group 2 (vasomotor symptoms absent)

Skin conductance DIAGNOSTIC_TEST

The sympathetic skin response recordings are performed with a four-channel electromyography apparatus (Nihon-Kohden). They are recorded from both sides of the sternum with the active and reference electrodes placed 2 cm apart from the midline.

Group 1 (vasomotor symptoms present); Group 2 (vasomotor symptoms absent)

ALU and LINE-1 DNA methylation analysis GENETIC

2 cc peripheric venous blood is drawn from the participants to the test tubes with EDTA. DNA isolation is done by Nucleospin Blood Kit (REF:740951.250, Macherey-Nagel). Epitect Fast DNA Bisulfite Kit (REF:59824, Qiagen) is used to do DNA bisulfite modification. ALU and LINE-1 site specific methylation primer is designed and the methylation patterns are compared between the groups by Epitect Methylight PCR Kit (Cat. No:59496, Qiagen).

Group 1 (vasomotor symptoms present); Group 2 (vasomotor symptoms absent)

The Menopause-Specific Quality of Life Questionnaire (MENQOL) DIAGNOSTIC_TEST

The Menopause-Specific Quality of Life Questionnaire (MENQOL) is filled by each participant to assess the health related quality of life before the polysomnography test.

Group 1 (vasomotor symptoms present); Group 2 (vasomotor symptoms absent)

Pittsburgh Sleep Quality Index (PSQI) DIAGNOSTIC_TEST

Pittsburgh Sleep Quality Index (PSQI) is done by each participant to assess the sleep quality and disturbances before thepolysomnography test.

Group 1 (vasomotor symptoms present); Group 2 (vasomotor symptoms absent)

Eligibility Criteria

• Age: 45 Years - 55 Years
• Sex: female
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64)
• Sampling Method: Non-Probability Sample

Study Population

The study population consists of women aged between 45-55 with low cardiovascular disease risk profile assessed by Framingham score system. The study population doesn't include any women who were already diagnosed with hypertension, coronary heart disease, diabetes, obstructive sleep apnea and stroke. It is a very homogenous population in terms of baseline cardiovascular disease risk apart from the presence or absence of hot flashes.

You may qualify if:

• years of age
• Women with serum FSH levels \>35 IU/L, serum estradiol levels \<20 pg/mL
• Women with low cardiovascular disease risk profile (Framingham score for coronary heart disease \<10%),
• Low high sensitive CRP levels (\<5 mg/L),
• Fasting glucose \<90 mg/dL,
• Fasting levels of insulin \<37.06 µIU/mL,
• Blood pressure \<140/90 mmHg (measured 2 times with 10 minutes interval),
• TSH \<4.2 mIU/L and fT4 \<1.7 ng/dL,
• Hemoglobin levels 12-16 g/dL, leucocyte count \<10.3\*10\^3/µL and neutrophil count \<4.9\*10\^3/µL; eosinophil count \<0.5\*10\^3/µL, basophil count \<0.2\*10\^3/µL

You may not qualify if:

• Women with high cardiovascular disease risk profile (Framingham score for coronary heart disease \>10%),
• Women who were diagnosed with a cardiovascular disease (coronary heart disease, stroke),
• Women with hypertension,
• Smokers,
• BMI \>30 kg/m2,
• Women with diabetes
• Women who were surgically postmenopausal,
• Women who used hormone therapy in the last three months,
• Women with obstructive sleep apnea
• Women who are using medicine which may cause sleep disturbances.

Sponsors & Collaborators

• Istanbul University - Cerrahpasa: lead
• Scientific Research Project Coordination Unit of Istanbul University-Cerrahpasa: collaborator

Study Sites (1)

Istanbul University-Cerrahpasa

Istanbul, 34098, Turkey (Türkiye)

Location

Biospecimen

Retention: SAMPLES WITH DNA

2 cc peripherical venous blood specimens were obtained from the participants into EDTA blood collection tubes. DNA was isolated by Nucleospin Blood Kit.

MeSH Terms

Conditions

Hot Flashes; Cardiovascular Diseases

Interventions

Polysomnography; Galvanic Skin Response

Condition Hierarchy (Ancestors)

Signs and Symptoms; Pathological Conditions, Signs and Symptoms

Intervention Hierarchy (Ancestors)

Monitoring, Physiologic; Diagnostic Techniques and Procedures; Diagnosis; Psychological Techniques; Investigative Techniques; Behavioral Disciplines and Activities; Electrophysiological Phenomena; Physiological Phenomena; Skin Physiological Phenomena; Integumentary System Physiological Phenomena

Study Officials

• Ipek Betul Ozcivit Erkan, MD

  Istanbul University - Cerrahpasa

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: observational
• Observational Model: COHORT
• Time Perspective: CROSS SECTIONAL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Resident Doctor

Study Record Dates

• First Submitted: May 9, 2023
• First Posted: June 7, 2023
• Study Start: December 1, 2020
• Primary Completion: December 1, 2022
• Study Completion (Estimated): March 8, 2032
• Last Updated: May 2, 2025

Record last verified: 2025-04

Locations

TU (1)