The Impact of Exercise Intervention on Biological and Functional Aging Through the Lens of Epigenetic Clocks
The Effects of Regular Exercise Training on Biological Aging in Older Adults: Investigation of Epigenetic and Functional Changes
This study investigated the effects of a structured 6-month high-intensity interval training (HIIT) program on blood DNA methylation-based measures of biological ageing, physical fitness, cardiovascular function, body composition, metabolic health, and the gut microbiome in adults aged 50-70 years.
Participants completed supervised exercise training three times per week for 6 months. Exercise intensity was individually prescribed based on estimated maximal heart rate (HRmax) and continuously monitored using Polar H10 heart-rate sensors. Perceived exertion was also assessed using the Borg Rating of Perceived Exertion scale to support safe and appropriate exercise intensity. The training program progressively increased the amount of high-intensity exercise over the intervention period. High-intensity intervals were performed above approximately 80% of estimated HRmax and were separated by 2-minute periods of active recovery.
Before and after the intervention, participants underwent physiological and functional assessments. Cardiovascular function was evaluated using echocardiography. Aerobic capacity was assessed using spiroergometry and the Chester Step Test for estimation of VO₂max. Muscular performance was evaluated using maximal vertical jump and handgrip strength tests. Exercise volume was quantified using oxygen-consumption and heart-rate data collected during exercise and expressed as MET-hours (MET-hrs), allowing the individual exercise dose accumulated during the intervention to be characterized.
Body composition was assessed using dual-energy X-ray absorptiometry (DEXA). Measurements included total and regional fat mass and fat-free mass, allowing changes in overall and regional body composition to be evaluated.
Blood samples were collected before and after the intervention, with participants refraining from exercise for 48 hours before blood collection. Samples were analyzed for selected metabolic and clinical biomarkers, including glucose, triglycerides, total cholesterol, HDL cholesterol, LDL cholesterol, and liver enzymes.
Blood samples were also used to assess blood DNA methylation-based measures of biological ageing. Genome-wide DNA methylation was measured using the Illumina Infinium MethylationEPIC BeadChip. Several DNA methylation-based ageing measures were calculated, including DNAmFitAge, DNAmPhenoAge, DNAmGrimAge, Hannum Age, DunedinPACE, and the Horvath Skin & Blood and pan-tissue clocks. Changes in biological age acceleration were evaluated between baseline and post-intervention.
The gut microbiome was characterized using shotgun metagenomic sequencing to assess changes in microbial composition and functional potential following the exercise intervention.
The primary overall objective of the study was to determine whether the individual volume of exercise performed, quantified as MET-hours, was associated with changes in blood DNA methylation-based biological age acceleration following the 6-month intervention. The study also examined whether exercise volume and training-related changes were associated with changes in physical fitness, cardiovascular function, body composition, metabolic biomarkers, and gut microbiome characteristics. The results may contribute to understanding whether the amount of regular high-intensity exercise is related to biological ageing and other physiological adaptations in 50< years old adults.
Study Overview
Status
Conditions
Intervention / Treatment
Study Type
Enrollment (Actual)
Phase
- Not Applicable
Contacts and Locations
Study Locations
-
-
Pest County
-
Budapest, Pest County, Hungary, 1123
- Research Center for Molecular Exercise Science, Hungarian University of Sports Science, Alkotás st. 42-48
-
-
Participation Criteria
Eligibility Criteria
Ages Eligible for Study
- Adult
- Older Adult
Accepts Healthy Volunteers
Description
Inclusion Criteria:
- Adults aged 50-70 years
- Legal capacity to provide informed consent
- Successful completion of a questionnaire-based and instrumental medical screening, confirming eligibility for physical exercise
- Physical activity level between 1 and 5 METs/day, estimated based on a questionnaire
Exclusion Criteria:
- Cardiovascular diseases that contraindicate physical exercise, including the presence of a pacemaker
- Musculoskeletal conditions that contraindicate physical exercise
- Acute or chronic kidney disease
- Acute or chronic liver disease
- Acute or chronic gastrointestinal disease
- Acute or chronic infectious diseases or febrile conditions
- Metabolic disorders or endocrine diseases
- Bleeding disorders or coagulation abnormalities
- Malignant diseases
- Autoimmune diseases
- Asthma
- Psychiatric or mental disorders
- Limited legal capacity or impaired ability to communicate
- Chronic alcohol consumption
Study Plan
How is the study designed?
Design Details
- Primary Purpose: Basic Science
- Allocation: N/A
- Interventional Model: Single Group Assignment
- Masking: None (Open Label)
Arms and Interventions
Participant Group / Arm |
Intervention / Treatment |
|---|---|
|
Experimental: Relative High Intensity Intervention Exercise Group for 6 month
Individuals over 50 years of age will undergo regular exercise training three times per week for 40-45 minutes using a high-intensity interval training (HIIT) modality for a period of 6 months.
Following health screening and a 2-3-week habituation period, exercise intensity will be monitored and prescribed using a chest-strap heart rate monitoring system.
Before and after the intervention, blood samples will be collected, and physiological and functional assessments will be conducted.
|
Participants completed supervised high-intensity interval training (HIIT) 3 times weekly for 6 months.
Exercise intensity was prescribed relative to estimated maximal heart rate (HRmax; Tanaka formula) and was monitored continuously using Polar H10 heart-rate sensors.
Sessions included a progressive warm-up, repeated 2-minute high-intensity intervals performed above 80% of HRmax, and 2-minute active recovery periods.
Perceived exertion was also monitored using the Borg 0-10 RPE scale, with an upper target of 8.
The protocol progressively increased training load: sessions 1-25 included five high-intensity intervals, sessions 26-45 six intervals, and sessions 46 onward seven intervals.
Session duration increased from 34 to 38 minutes in the final phase.
|
What is the study measuring?
Primary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
|
Epigenetic Age Accelration
Time Frame: Whole blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
Whole blood samples were collected and analyzed using the Illumina EPIC BeadChip assay to quantify DNA methylation at specific CpG sites.
DNA methylation-based biological age estimators, including DNAmFitAge, DNAmPhenoAge, and DNAmGrimAge, were calculated from the methylation data.
Age acceleration values were expressed in years, and changes in these measures were correlated with changes in other primary and secondary outcome measures.
|
Whole blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
|
Relative aerobic capaity spiroergometry: VO2 max measurement
Time Frame: Spiriergomerty was conducted within 2 weeks before the first exercise session as the baseline measurement and after at least 48 hours, but no more than 14 days, after the final exercise session.
|
Relative aerobic capacity was evaluated using cardiopulmonary exercise testing (spiroergometry) and expressed as ml/kg/min
|
Spiriergomerty was conducted within 2 weeks before the first exercise session as the baseline measurement and after at least 48 hours, but no more than 14 days, after the final exercise session.
|
|
Relative aerobic capacity esimation: Chester step test
Time Frame: The Chester Step Test was performed at the first exercise session, which served as the baseline assessment, and was repeated monthly thereafter to monitor changes in aerobic capacity throughout the intervention period.
|
The Chester Step Test was conducted to estimate changes in VO₂max during the 6-month intervention period.
Heart rate responses recorded during each assessment were used to estimate relative aerobic capacity, expressed as mL/kg/min.
|
The Chester Step Test was performed at the first exercise session, which served as the baseline assessment, and was repeated monthly thereafter to monitor changes in aerobic capacity throughout the intervention period.
|
|
Exercise Volume: MET-hours
Time Frame: Heart rate values and exercise duration were recorded during each exercise session throughout the 6-month intervention period.
|
Exercise volume was calculated based on estimated VO₂max, average and resting heart rate during exercise, and exercise duration.
Exercise volume was expressed as MET-hours.
|
Heart rate values and exercise duration were recorded during each exercise session throughout the 6-month intervention period.
|
|
Total body muscular power
Time Frame: Grip strength was measured within 2 weeks before the first exercise session as the baseline measurement. The second measurement was collected at least 48 hours, but no more than 14 days, after the final exercise session.
|
Total body muscular strength was measured using a handheld dynamometer, and grip force was expressed in kilograms (kg).
|
Grip strength was measured within 2 weeks before the first exercise session as the baseline measurement. The second measurement was collected at least 48 hours, but no more than 14 days, after the final exercise session.
|
|
Echocardiographic measurements
Time Frame: Echocardiographic data was collected within 2 weeks before the first exercise session as the baseline measurement. The second second measurement was conducted at least 48 hours, but no more than 5 days, after the final exercise session.
|
Echocardiographic measurements included assessments of cardiac morphology and structure, as well as parameters of cardiac function.
These measurements were used to characterize structural and functional cardiac adaptations and to evaluate changes in cardiovascular function over the course of the intervention.
|
Echocardiographic data was collected within 2 weeks before the first exercise session as the baseline measurement. The second second measurement was conducted at least 48 hours, but no more than 5 days, after the final exercise session.
|
Secondary Outcome Measures
Outcome Measure |
Measure Description |
Time Frame |
|---|---|---|
|
Blood chemistry: LDL
Time Frame: Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
Low-density lipoprotein cholesterol (LDL-C) was measured from venous blood reum samples as a marker of lipid metabolism and cardiovascular risk.
LDL-C concentration was expressed in mmol/L.
|
Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
|
Blood chemistry: HDL
Time Frame: Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
High-density lipoprotein cholesterol (HDL) was measured from venous blood samples as a marker of lipid metabolism and cardiovascular health.
HDL-C concentration was expressed in mmol/L.
|
Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
|
Blood chemistry: Fastung glucose
Time Frame: Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
Fasting blood glucose was measured from venous blood samples following an overnight fast as a marker of glucose metabolism.
Glucose concentration was expressed in mmol/L.
|
Blood was collected within 2 weeks before the first exercise session as the baseline measurement. The second blood sample was collected at least 48 hours, but no more than 5 days, after the final exercise session.
|
|
Fecal microbial status
Time Frame: Fecal samples were collected within 2 weeks before the first exercise session as the baseline measurement. The second fecal sample was collected at least 48 hours, but no more than 2 weeks, after the final exercise session.
|
Gut microbiome composition and functional characteristics were assessed from fecal samples using shotgun metagenomic sequencing before and after the intervention.
Microbiome composition was characterized at the species level, with individual taxa expressed as relative abundance.
|
Fecal samples were collected within 2 weeks before the first exercise session as the baseline measurement. The second fecal sample was collected at least 48 hours, but no more than 2 weeks, after the final exercise session.
|
Collaborators and Investigators
Collaborators
Study record dates
Study Major Dates
Study Start (Actual)
Primary Completion (Actual)
Study Completion (Actual)
Study Registration Dates
First Submitted
First Submitted That Met QC Criteria
First Posted (Actual)
Study Record Updates
Last Update Posted (Actual)
Last Update Submitted That Met QC Criteria
Last Verified
More Information
Terms related to this study
Keywords
Other Study ID Numbers
- NNGYK/14120-7/2025_FEP
- 152986 (Other Grant/Funding Number: NKFIH under the National Research Excellence Programme - STARTING_25)
Plan for Individual participant data (IPD)
Plan to Share Individual Participant Data (IPD)?
Drug and device information, study documents
Studies a U.S. FDA-regulated drug product
Studies a U.S. FDA-regulated device product
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.