The Impact of Exercise Intervention on Biological and Functional Aging Through the Lens of Epigenetic Clocks

NCT07799103 | Completed

• 2 other identifiers: NNGYK/14120-7/2025_FEP152986
• Study Type: interventional
• Target: 74
• Locations: 1 country
• Sites: 1

Brief Summary

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.

Conditions

Age Over 50 Years

Keywords

epigentic; biological aging; exersice

Outcome Measures

Primary Outcomes (6)

• Epigenetic Age Accelration

  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

  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

  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

  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

  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

  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 Outcomes (4)

• Blood chemistry: LDL

  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

  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

  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

  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.

Study Arms (1)

Relative High Intensity Intervention Exercise Group for 6 month

EXPERIMENTAL

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.

Other: Exercise - Relative High-Intensity Interval Training (HIIT)

Interventions

Exercise - Relative High-Intensity Interval Training (HIIT) OTHER

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.

Relative High Intensity Intervention Exercise Group for 6 month

Eligibility Criteria

• Age: 50 Years - 70 Years
• Healthy Volunteers: Yes
• Age Groups: Adult (18-64), Older Adult (65+)

You may qualify if:

• 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

You may not qualify if:

• 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

Sponsors & Collaborators

• Hungarian University of Sports Science: lead
• Semmelweis University: collaborator

Study Sites (1)

Research Center for Molecular Exercise Science, Hungarian University of Sports Science, Alkotás st. 42-48

Budapest, Pest County, 1123, Hungary

Location

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: NA
• Masking: NONE
• Purpose: BASIC SCIENCE
• Intervention Model: SINGLE GROUP
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Senior Researcher

Study Record Dates

• First Submitted: August 26, 2026
• First Posted: September 2, 2026
• Study Start: May 1, 2022
• Primary Completion: May 1, 2024
• Study Completion: June 30, 2024
• Last Updated: September 2, 2026

Record last verified: 2026-08

Locations

HU (1)