Does Human Skeletal Muscle Possess an Epigenetic Memory of Testosterone?

Study Overview
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Brief Summary
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This project's primary aim of this double-blinded, randomised, placebo-controlled trial is to investigate whether short-term testosterone administration +/- resistance exercise training induces a muscle memory response that can lead to longer-lasting benefits in aged human skeletal muscle. The investigators will provide older men with the anabolic hormone, testosterone or placebo, with or without resistance training, followed by a period of testosterone abstinence and detraining, followed by a subsequent repeated period of resistance training (retraining). This will help determine if earlier encounters with short-term testosterone administration can be remembered and if adaptation to later retraining can be enhanced as a consequence of encountering testosterone earlier.

Official Title
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Does Human Skeletal Muscle Possess an Epigenetic Memory of Testosterone?

Conditions
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Healthy Aging, Age-Related Sarcopenia, Testosterone Deficiency

Intervention / Treatment
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* Drug: Saline
* Drug: Testosterone Undecanoate
* Drug: Saline + Resistance exercise training
* Drug: Testosterone Undecanoate + Resistance exercise training


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Sedentary males 55-70 years old Serum testosterone levels >8 nmol/L measured in the morning Without any known illness, disease or other conditions Undergone screening through medical questionnaire, physical examination, routine blood tests and urine sample Written informed consent received Exclusion Criteria: Current or previous participation in a formal exercise regime A BMI < 18 or > 30 kg·m2 Hypersensitivity to the study drug or to any of its constituents Active cardiovascular disease: uncontrolled hypertension (BP > 160/100 mmHg), angina, heart failure (class III/IV), arrhythmia, right to left cardiac shunt, recent cardiac event Family history of early (<55y) death from cardiovascular disease Haematocrit >50% Malignancy Prostate-specific antigen (PSA) >4 ng/mL Lower urinary tract symptoms Taking beta-adrenergic blocking agents, statins, non-steroidal anti-inflammatory drugs Cerebrovascular disease: previous stroke, aneurysm (large vessel or intracranial), epilepsy Respiratory diseases including: pulmonary hypertension, chronic obstructive pulmanary disease (COPD), asthma, sleep apnoea Metabolic disease: hyper and hypo parathyroidism, untreated hyper and hypothyroidism, Cushing's disease, type 1 or 2 diabetes Active inflammatory bowel or renal disease Current or previous steroid treatment or hormone replacement therapy Clotting dysfunction Musculoskeletal or neurological disorders Alcohol or drug abuse Receiving oral anticoagulants Serum testosterone levels above the reference range for 50 year olds (>32 nmol/L) (Bjerner et al., 2009) measured in the morning 1

Ages Eligible for Study
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Minimum Age: 55 Years
Maximum Age: 70 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Placebo<br>The placebo group will complete 10-week treatment period where they continue with their regular habitual daily physical activity and receive two placebo (saline) injections (at baseline and week 3). They will then undergo a 12-week period with no treatment and no training, where they just do their regular habitual daily physical activity. Before they undertake a period of structured, progressive resistance training for 10-weeks. Questionnaires, physiological and psychological measures, skeletal muscle biopsies and blood samples will be taken at time points of: Baseline (week 0) Treatment period (week 10) Detraining and placebo abstinence (week 22) Retraining (week 32) | Drug: Saline<br>* Two placebo injections one at baseline and one week 3.<br>* Other names: Placebo;|
| Experimental: Testosterone Undecanoate<br>The testosterone group will complete 10-week treatment period where they continue with their regular habitual daily physical activity and receive two testosterone undecanoate (Nebido) injections (1000 mg/4 ml at baseline and 500 mg/2 ml week 3). They will then undergo a 12-week period with no treatment and no training, where they just do their regular habitual daily physical activity. Before they undertake a period of structured, progressive resistance training for 10-weeks. Questionnaires, physiological and psychological measures, skeletal muscle biopsies and blood samples will be taken at time points of: Baseline (week 0) Treatment period (week 10) Detraining and testosterone abstinence (week 22) Retraining (week 32) | Drug: Testosterone Undecanoate<br>* Two testosterone undecanoate injections, 1000 mg/4 ml at baseline, 500 mg/2 ml at week 3.<br>* Other names: Nebido, Grünenthal (Grünenthal Norway AS);|
| Placebo Comparator: Resistance exercise training + Placebo<br>The resistance exercise training + placebo group will complete 10-week treatment period where they undergo a period of structured, progressive resistance training and receive two placebo (saline) injections (at baseline and week 3). They will then undergo a 12-week period with no treatment and no training, where they return to their regular habitual daily physical activity. Before they undertake a second period of structured, progressive resistance training for 10-weeks. Questionnaires, physiological and psychological measures, skeletal muscle biopsies and blood samples will be taken at time points of: Baseline (week 0) Treatment period (week 10) Detraining and placebo abstinence (week 22) Retraining (week 32) | Drug: Saline + Resistance exercise training<br>* Two placebo injections one at baseline and one week 3 and10 weeks of supervised, structured, progressive resistance training.<br>* Other names: Placebo + Resistance exercise training;|
| Experimental: Resistance exercise training + Testosterone Undecanoate<br>The resistance exercise training + testosterone group will complete 10-week treatment period where they undergo a period of structured, progressive resistance training and receive two testosterone undecanoate (Nebido) injections (1000 mg/4 ml at baseline and 500 mg/2 ml week 3). They will then undergo a 12-week period with no treatment and no training, where they return to their regular habitual daily physical activity. Before they undertake a second period of structured, progressive resistance training for 10-weeks. Questionnaires, physiological and psychological measures, skeletal muscle biopsies and blood samples will be taken at time points of: Baseline (week 0) Treatment period (week 10) Detraining and testosterone abstinence (week 22) Retraining (week 32) | Drug: Testosterone Undecanoate + Resistance exercise training<br>* Two testosterone undecanoate injections, 1000 mg/4 ml at baseline, 500 mg/2 ml at week 3 and 10 weeks of supervised, structured, progressive resistance training.<br>* Other names: Nebido, Grünenthal (Grünenthal Norway AS) + Resistance exercise training;|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Fat-free mass | Change and differences in fat-free mass (g) measured by dual x-ray absorptiometry (DEXA). | Baseline and weeks 10, 22, 32 | 
| Skeletal muscle size and cross-sectional area (CSA) | Change and differences in skeletal muscle size and CSA measured by ultrasound | Baseline and week 5,10, 16, 22, 27, 32 | 
| Skeletal muscle fibre CSA | Change and differences in skeletal muscle fibre CSA measure determined by immunohistochemistry | Baseline and weeks 10, 22, 32 | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| DNA methylation in skeletal muscle and blood | Methylation measured in difference/fold change values relative to appropriate controls. | Baseline and weeks 10, 22, 32 | 
| Gene expression in skeletal muscle and blood | Gene expression measured in difference/fold change values relative to appropriate controls. | Baseline and weeks 10, 22, 32 | 
| Myonuclei | Change and differences in number of myonuclei determined by immunohistochemistry | Baseline and weeks 10, 22, 32 | 
| Satellite cells | Change and differences in number of satellite cells determined by immunohistochemistry | Baseline and weeks 10, 22, 32 | 
| Isometric muscle strength | Change and differences in peak muscle strength (N) using isokinetic dynamometry | Baseline and weeks 5, 10, 16, 22, 27, 32 | 
| Dynamic muscle strength | Change and differences in 1-repetition maximum | Baseline and weeks 5, 10, 16, 22, 27, 32 | 
| Muscle force-velocity profiling | Change and differences in force (N) and velocity (m/s) derived from a 10-repetition FV-test | Baseline and weeks 5, 10, 16, 22, 27, 32 | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Healthy aging, Preventing Age-related Sarcopenia, Overcoming Anabolic Resistance, Anti-Doping