Physical InactivityNCT07771075
High-Intensity Interval Training and Monocyte Epigenetic Adaptation After Detraining
A study testing High-Intensity Interval Training and Moderate-Intensity Continuous Training in people with Physical Inactivity, run by Saif Rasheed Ghanim at 1 site in Iraq.
Trial summary
This randomized, work-matched training-detraining-rechallenge study evaluated whether repeated high-intensity interval training produces persistent functional and epigenetic adaptations in human monocytes. Read more about this study
- Who is it for?
- Ages 18 to 30 yearsHealthy volunteers can take part
- Conditions the study is for
- Physical Inactivity
- What is being tested?
- High-Intensity Interval Training and Moderate-Intensity Continuous Training
- Type of treatment
- Lifestyle or therapy programme
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 80 participants
- Where
- 1 siteIraq
- Study dates
- 14 Jan 2025to 2 Jul 2025
- Phase
- Interventional
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 18 to 30 years
- All sexes
- Healthy volunteers accepted
- Adult
Who may be able to join
- Men and women aged 18-30 years. Healthy adults. Physically inactive, defined as participating in less than 150 minutes per week of structured moderate-to-vigorous exercise during the preceding 6 months.
Who cannot join
- Major cardiovascular, metabolic, respiratory, renal, endocrine, inflammatory, autoimmune, or hematological disease.
- Current tobacco use. Current use of relevant immunomodulatory, anti-inflammatory, corticosteroid, or cardiovascular medications.
- Acute infection within 4 weeks before baseline assessment. Musculoskeletal contraindications to maximal cycling exercise. Pregnancy or lactation.
Trial locations and contacts
1 site in 1 country.
Iraq1 site
Exercise Physiology Laboratory, College of Physical Education and Sports Sciences, University of Samarra
Samarra, Muhafazat Salah ad Din
About this study
Official title: Repeated High-Intensity Exercise Induces Persistent H3K18 Lactylation and Trained Immunity in Human Monocytes: A Randomized Work-Matched Training-Detraining-Rechallenge Trial
This randomized, work-matched training-detraining-rechallenge study evaluated whether repeated high-intensity interval training produces persistent functional and epigenetic adaptations in human monocytes. Eighty healthy physically inactive adults aged 18-30 years were randomly assigned in a 2:2:1 ratio to high-intensity interval training (HIIT), moderate-intensity continuous training (MCT), or a non-training control group. HIIT and MCT completed 6 weeks of supervised cycling training matched for external mechanical work, followed by 4 weeks without prescribed structured training. All groups subsequently completed a standardized 30-minute cycling rechallenge. The primary outcome was the between-group difference in change in ex-vivo LPS-stimulated TNF-alpha release from baseline to the post-detraining assessment. Secondary outcomes included IL-6 release, H3K18 lactylation, monocyte subset distribution, blood lactate exposure, chromatin accessibility, H3K18la CUT\&Tag occupancy, and RNA-seq transcriptional profiling.
Read the detailed description
Participants completed baseline cardiopulmonary exercise testing to determine peak oxygen uptake (VO2peak) and maximal aerobic power (MAP). Eligible participants were randomized to HIIT, MCT, or Control. The HIIT intervention consisted of 4 × 4-minute cycling intervals at 85% MAP separated by active recovery at 40% MAP, with standardized warm-up and cool-down periods. The MCT intervention consisted of continuous cycling at 55% MAP for a duration selected to match the external mechanical work of the HIIT sessions. Both exercise groups trained three times per week for 6 weeks.
Blood and monocyte assessments were performed at baseline (T0), 72 hours after the final training session (T1), after 4 weeks without prescribed structured training (T2_PRE), and 120 minutes after a standardized 30-minute cycling rechallenge at 60% of baseline MAP (T3_POST).
Classical monocytes were isolated and stimulated ex vivo with ultrapure Escherichia coli O111:B4 lipopolysaccharide (10 ng/mL) for 24 hours. TNF-alpha was the prespecified primary functional outcome, with IL-6 as a key secondary outcome. Epigenetic and molecular analyses included global H3K18 lactylation, ATAC-seq chromatin accessibility profiling, H3K18la CUT\&Tag, and RNA-seq. Blood lactate kinetics and cumulative lactate exposure across the training intervention were also assessed.
The primary comparison was HIIT versus MCT. The Control group served as a supportive non-training reference. Longitudinal outcomes were analyzed using mixed models for repeated measures under the intention-to-treat principle.
- Conditions
- Physical Inactivity
- Being tested
- High-Intensity Interval Training, Moderate-Intensity Continuous Training
- Keywords
- High-Intensity Interval Training, Histone Lactylation, H3K18la, Monocytes, Trained Immunity, Innate Immunity, Blood Lactate, Epigenetics
- Medical subject headings
- Sedentary Behavior
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
High-Intensity Interval TrainingBehavioural
Supervised cycling HIIT performed three sessions per week for 6 weeks. Each session included a 5-minute warm-up at 30% MAP, four 4-minute intervals at 85% MAP separated by three 3-minute active recovery periods at 40% MAP, and a 5-minute cool-down at 30% MAP.
Moderate-Intensity Continuous TrainingBehavioural
Supervised moderate-intensity continuous cycling performed three sessions per week for 6 weeks. Each session included a 5-minute warm-up at 30% MAP, 31 minutes and 16 seconds of continuous cycling at 55% MAP, and a 5-minute cool-down at 30% MAP. The protocol was designed to match the external mechanical work performed by the HIIT group.
| Study group | Role | What they receive |
|---|---|---|
| High-Intensity Interval Training | Experimental | Participants completed 6 weeks of supervised cycling high-intensity interval training, three sessions per week. Each session consisted of a 5-minute warm-up at 30% of maximal aerobic power (MAP), four 4-minute intervals at 85% MAP separated by three 3-minute active recovery periods at 40% MAP, followed by a 5-minute cool-down at 30% MAP. Receives: Behavioral: High-Intensity Interval Training |
| Moderate-Intensity Continuous Training | Active comparator | Participants completed 6 weeks of supervised moderate-intensity continuous cycling training, three sessions per week. Each session consisted of a 5-minute warm-up at 30% of maximal aerobic power (MAP), 31 minutes and 16 seconds of continuous cycling at 55% MAP, followed by a 5-minute cool-down at 30% MAP. The protocol was designed to match the external mechanical work performed by the HIIT group. Receives: Behavioral: Moderate-Intensity Continuous Training |
| Control | No intervention | Participants received no prescribed structured exercise training during the 6-week intervention period and were instructed to maintain their habitual physical activity and lifestyle. They underwent the same study assessments as the exercise groups, including the 4-week detraining period and the standardized 30-minute cycling rechallenge. |
What is being measured
The primary outcome is the main question the study is designed to answer.
Change in LPS-Stimulated TNF-Alpha Release From Classical Monocytes
TNF-alpha concentration (pg/mL) was measured in culture supernatants after 24-hour ex-vivo stimulation of purified classical CD14++CD16- monocytes with 10 ng/mL ultrapure Escherichia coli O111:B4 lipopolysaccharide. The primary outcome was the change from baseline (T0) to the post-detraining assessment (T2_PRE), with the primary comparison between the HIIT and MCT groups.
Measured: Baseline (T0) and 4 weeks after completion of the 6-week training intervention (T2_PRE).
Secondary outcomes (7)
Change in LPS-Stimulated IL-6 Release From Classical Monocytes
IL-6 concentration (pg/mL) was measured in culture supernatants after 24-hour ex-vivo stimulation of purified classical CD14++CD16- monocytes with 10 ng/mL ultrapure Escherichia coli O111:B4 lipopolysaccharide. The outcome was the change from baseline (T0) to the post-detraining assessment (T2_PRE), with comparison between the HIIT and MCT groups.
Measured: Baseline (T0) and 4 weeks after completion of the 6-week training intervention (T2_PRE).
Exercise-Rechallenge Change in LPS-Stimulated TNF-Alpha Release From Classical Monocytes
TNF-alpha concentration (pg/mL) was measured after 24-hour ex-vivo stimulation of purified classical CD14++CD16- monocytes with 10 ng/mL ultrapure Escherichia coli O111:B4 lipopolysaccharide. The outcome was the within-participant change from the post-detraining pre-rechallenge assessment (T2_PRE) to 120 minutes after the standardized exercise rechallenge (T3_POST), with the principal comparison between the HIIT and MCT groups.
Measured: Immediately before the standardized exercise rechallenge (T2_PRE) and 120 minutes after the rechallenge (T3_POST).
Exercise-Rechallenge Change in LPS-Stimulated IL-6 Release From Classical Monocytes
IL-6 concentration (pg/mL) was measured after 24-hour ex-vivo stimulation of purified classical CD14++CD16- monocytes with 10 ng/mL ultrapure Escherichia coli O111:B4 lipopolysaccharide. The outcome was the within-participant change from the post-detraining pre-rechallenge assessment (T2_PRE) to 120 minutes after the standardized exercise rechallenge (T3_POST), with the principal comparison between the HIIT and MCT groups.
Measured: Immediately before the standardized exercise rechallenge (T2_PRE) and 120 minutes after the rechallenge (T3_POST).
Change in Monocyte H3K18 Lactylation Across Study Time Points
Histone H3 lysine 18 lactylation (H3K18la) was quantified in purified classical CD14++CD16- monocytes and normalized to total histone H3 abundance. H3K18la was assessed to determine training-induced elevation, persistence after 4 weeks without prescribed structured training, and re-amplification following the standardized exercise rechallenge.
Measured: Baseline (T0), 72 hours after the final training session (T1), 4 weeks after training cessation before rechallenge (T2_PRE), and 120 minutes after the standardized exercise rechallenge (T3_POST).
Genome-Wide Chromatin Accessibility in Classical Monocytes
Genome-wide chromatin accessibility was assessed by ATAC-seq in purified classical CD14++CD16- monocytes. Differentially accessible regions between the HIIT and MCT groups were identified using a false discovery rate (FDR) <0.05 and an absolute log2 fold-change >1.0.
Measured: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Genome-Wide H3K18la Occupancy in Classical Monocytes
Genome-wide histone H3 lysine 18 lactylation (H3K18la) occupancy was profiled by CUT\&Tag in purified classical CD14++CD16- monocytes. H3K18la-enriched genomic regions were identified and their overlap with differentially accessible ATAC-seq regions was evaluated between the HIIT and MCT groups.
Measured: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Genome-Wide Differential Gene Expression in Classical Monocytes
Genome-wide transcript abundance was assessed by RNA sequencing (RNA-seq) in purified classical CD14++CD16- monocytes. Differential gene expression between the HIIT and MCT groups was evaluated using DESeq2, with differentially expressed genes defined using a false discovery rate (FDR) <0.05 and an absolute log2 fold-change >1.0.
Measured: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted18 Aug 2026
- Study start14 Jan 2025
- Primary completion21 Jun 2025
- Study completion2 Jul 2025
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Other trials for Physical InactivityStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Basic science
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Model description
- Participants were randomized in a 2:2:1 ratio to HIIT, MCT, or a non-training Control group and were followed in parallel through the training, detraining, and standardized exercise-rechallenge phases.
- Masking
- Single blind
- Who is masked
- Outcomes assessor
- Enrolment
- 80 participants (actual)
Sponsor and investigators
- Lead sponsor
- Saif Rasheed Ghanim
- Sponsor type
- University, hospital or other
- Responsible party
- Sponsor-investigator, Saif Rasheed Ghanim, Samarra University
- Investigators
- Saif Rasheed Ghanim, PhD, Principal investigator, University of Samarra, College of Physical Education and Sports Sciences
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT07771075
- Sponsor's protocol ID
- SRG-HIIT-H3K18LA-2026-01
- Status last verified
- Aug 2026
- First submitted
- 13 Aug 2026
- Last updated
- 22 Sep 2026
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- Yes: De-identified individual participant data underlying the results reported in the primary publication will be available for sharing. Shared data may include participant characteristics, intervention allocation, longitudinal cytokine outcomes, H3K18 lactylation measures, monocyte phenotyping, and processed omics-derived variables, after removal of information that could permit participant identification.
Study at a glance
From the registry record, last updated by the sponsor on 22 Sep 2026. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Repeated High-Intensity Exercise Induces Persistent H3K18 Lactylation and Trained Immunity in Human Monocytes: A Randomized Work-Matched Training-Detraining-Rechallenge Trial
- ClinicalTrials.gov ID
- NCT07771075
- Acronym / sponsor ID
- SRG-HIIT-H3K18LA-2026-01
- Conditions
- Physical Inactivity
- Keywords
- High-Intensity Interval Training, Histone Lactylation, H3K18la, Monocytes, Trained Immunity, Innate Immunity, Blood Lactate, Epigenetics
- Study type
- Interventional
- Recruitment status
- Completed
- Enrolment
- 80 participants (actual)
- Lead sponsor
- Saif Rasheed GhanimUniversity, hospital or other
- Study start
- 14 Jan 2025 (actual)
- Primary completion
- 21 Jun 2025 (actual)
- Study completion
- 2 Jul 2025 (actual)
- First posted
- 18 Aug 2026 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
This randomized, work-matched training-detraining-rechallenge study evaluated whether repeated high-intensity interval training produces persistent functional and epigenetic adaptations in human monocytes. Eighty healthy physically inactive adults aged 18-30 years were randomly assigned in a 2:2:1 ratio to high-intensity interval training (HIIT), moderate-intensity continuous training (MCT), or a non-training control group. HIIT and MCT completed 6 weeks of supervised cycling training matched for external mechanical work, followed by 4 weeks without prescribed structured training. All groups subsequently completed a standardized 30-minute cycling rechallenge. The primary outcome was the between-group difference in change in ex-vivo LPS-stimulated TNF-alpha release from baseline to the post-detraining assessment. Secondary outcomes included IL-6 release, H3K18 lactylation, monocyte subset distribution, blood lactate exposure, chromatin accessibility, H3K18la CUT\&Tag occupancy, and RNA-seq transcriptional profiling.
Detailed description
Participants completed baseline cardiopulmonary exercise testing to determine peak oxygen uptake (VO2peak) and maximal aerobic power (MAP). Eligible participants were randomized to HIIT, MCT, or Control. The HIIT intervention consisted of 4 × 4-minute cycling intervals at 85% MAP separated by active recovery at 40% MAP, with standardized warm-up and cool-down periods. The MCT intervention consisted of continuous cycling at 55% MAP for a duration selected to match the external mechanical work of the HIIT sessions. Both exercise groups trained three times per week for 6 weeks.
Blood and monocyte assessments were performed at baseline (T0), 72 hours after the final training session (T1), after 4 weeks without prescribed structured training (T2_PRE), and 120 minutes after a standardized 30-minute cycling rechallenge at 60% of baseline MAP (T3_POST).
Classical monocytes were isolated and stimulated ex vivo with ultrapure Escherichia coli O111:B4 lipopolysaccharide (10 ng/mL) for 24 hours. TNF-alpha was the prespecified primary functional outcome, with IL-6 as a key secondary outcome. Epigenetic and molecular analyses included global H3K18 lactylation, ATAC-seq chromatin accessibility profiling, H3K18la CUT\&Tag, and RNA-seq. Blood lactate kinetics and cumulative lactate exposure across the training intervention were also assessed.
The primary comparison was HIIT versus MCT. The Control group served as a supportive non-training reference. Longitudinal outcomes were analyzed using mixed models for repeated measures under the intention-to-treat principle.
Study design
- Primary purpose
- Basic science
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Masking
- Single blind (outcomes assessor)
- Number of arms
- 3
- Model description
- Participants were randomized in a 2:2:1 ratio to HIIT, MCT, or a non-training Control group and were followed in parallel through the training, detraining, and standardized exercise-rechallenge phases.
| Arm | Type | Receives |
|---|---|---|
| High-Intensity Interval Training | Experimental | Behavioral: High-Intensity Interval TrainingParticipants completed 6 weeks of supervised cycling high-intensity interval training, three sessions per week. Each session consisted of a 5-minute warm-up at 30% of maximal aerobic power (MAP), four 4-minute intervals at 85% MAP separated by three 3-minute active recovery periods at 40% MAP, followed by a 5-minute cool-down at 30% MAP. |
| Moderate-Intensity Continuous Training | Active comparator | Behavioral: Moderate-Intensity Continuous TrainingParticipants completed 6 weeks of supervised moderate-intensity continuous cycling training, three sessions per week. Each session consisted of a 5-minute warm-up at 30% of maximal aerobic power (MAP), 31 minutes and 16 seconds of continuous cycling at 55% MAP, followed by a 5-minute cool-down at 30% MAP. The protocol was designed to match the external mechanical work performed by the HIIT group. |
| Control | No intervention | –Participants received no prescribed structured exercise training during the 6-week intervention period and were instructed to maintain their habitual physical activity and lifestyle. They underwent the same study assessments as the exercise groups, including the 4-week detraining period and the standardized 30-minute cycling rechallenge. |
Who may qualify
- Ages
- 18 to 30 years
- Sex
- All
- Healthy volunteers
- Accepted
- Age groups
- Adult
Key inclusion criteria (1 listed)
- Men and women aged 18-30 years. Healthy adults. Physically inactive, defined as participating in less than 150 minutes per week of structured moderate-to-vigorous exercise during the preceding 6 months.
Key exclusion criteria (3 listed)
- Major cardiovascular, metabolic, respiratory, renal, endocrine, inflammatory, autoimmune, or hematological disease.
- Current tobacco use. Current use of relevant immunomodulatory, anti-inflammatory, corticosteroid, or cardiovascular medications.
- Acute infection within 4 weeks before baseline assessment. Musculoskeletal contraindications to maximal cycling exercise. Pregnancy or lactation.
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| High-Intensity Interval TrainingSupervised cycling HIIT performed three sessions per week for 6 weeks. Each session included a 5-minute warm-up at 30% MAP, four 4-minute intervals at 85% MAP separated by three 3-minute active recovery periods at 40% MAP, and a 5-minute cool-down at 30% MAP. | Behavioural | High-Intensity Interval Training |
| Moderate-Intensity Continuous TrainingSupervised moderate-intensity continuous cycling performed three sessions per week for 6 weeks. Each session included a 5-minute warm-up at 30% MAP, 31 minutes and 16 seconds of continuous cycling at 55% MAP, and a 5-minute cool-down at 30% MAP. The protocol was designed to match the external mechanical work performed by the HIIT group. | Behavioural | Moderate-Intensity Continuous Training |
Primary outcome
Change in LPS-Stimulated TNF-Alpha Release From Classical Monocytes
Time frame: Baseline (T0) and 4 weeks after completion of the 6-week training intervention (T2_PRE).
TNF-alpha concentration (pg/mL) was measured in culture supernatants after 24-hour ex-vivo stimulation of purified classical CD14++CD16- monocytes with 10 ng/mL ultrapure Escherichia coli O111:B4 lipopolysaccharide. The primary outcome was the change from baseline (T0) to the post-detraining assessment (T2_PRE), with the primary comparison between the HIIT and MCT groups.
7 secondary outcomes
Change in LPS-Stimulated IL-6 Release From Classical Monocytes
Time frame: Baseline (T0) and 4 weeks after completion of the 6-week training intervention (T2_PRE).
Exercise-Rechallenge Change in LPS-Stimulated TNF-Alpha Release From Classical Monocytes
Time frame: Immediately before the standardized exercise rechallenge (T2_PRE) and 120 minutes after the rechallenge (T3_POST).
Exercise-Rechallenge Change in LPS-Stimulated IL-6 Release From Classical Monocytes
Time frame: Immediately before the standardized exercise rechallenge (T2_PRE) and 120 minutes after the rechallenge (T3_POST).
Change in Monocyte H3K18 Lactylation Across Study Time Points
Time frame: Baseline (T0), 72 hours after the final training session (T1), 4 weeks after training cessation before rechallenge (T2_PRE), and 120 minutes after the standardized exercise rechallenge (T3_POST).
Genome-Wide Chromatin Accessibility in Classical Monocytes
Time frame: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Genome-Wide H3K18la Occupancy in Classical Monocytes
Time frame: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Genome-Wide Differential Gene Expression in Classical Monocytes
Time frame: Four weeks after completion of the 6-week training intervention, before the standardized exercise rechallenge (T2_PRE).
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 22 Sep 2026.
- Individual participant dataSharing planned: Yes. De-identified individual participant data underlying the results reported in the primary publication will be available for sharing. Shared data may include participant characteristics, intervention allocation, longitudinal cytokine outcomes, H3K18 lactylation measures, monocyte phenotyping, and processed omics-derived variables, after removal of information that could permit participant identification.
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
Saif Rasheed Ghanim, PhD principal investigator
University of Samarra, College of Physical Education and Sports Sciences
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing High-Intensity Interval Training and Moderate-Intensity Continuous Training in people with Physical Inactivity, run by Saif Rasheed Ghanim at 1 site in Iraq.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 1 inclusion and 3 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 22 Sep 2026. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Iraq. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Change in LPS-Stimulated TNF-Alpha Release From Classical Monocytes (time frame: Baseline (T0) and 4 weeks after completion of the 6-week training intervention (T2_PRE).). It is what the study was designed to measure, not a finding.
Where can I read the original study information?
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 22 Sep 2026.