mTOR as Mediator of Insulin Sensitivity Study

Study Overview
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Brief Summary
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The study investigates the role of mTOR in mediating enhancement of muscle insulin sensitivity following a single bout of exercise. This will be investigated in young healthy male subjects by administering the pharmacological mTOR inhibitor Rapamycin in a crossover blinded experimental setup known to enhance muscle insulin sensitivity following one-legged knee-extensor exercise.

Detailed Description
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Trial A: During trial A the study subject will arrive at the research laboratory and eat a morning meal at 6:30 a.m. corresponding to 5% of the daily energy requirements. It is requested that the study subject will arrive in a car or with public transportation. At the same time, the subject will ingest 16 mg of Rapamycin or placebo (800 mg Calcium) in pill form. Following ingestion, the study subject will rest for 2 hour and at that time catheters are inserted in both femoral veins, the femoral artery in one of the legs and in each forearm vein (antecubital veins) for later blood sampling and intravenous infusion of insulin, glucose, stable D2-glucose and stable 13C6-phenylalanine. At 8:30 a.m. the study subjects will perform one-leg knee extension exercise for 1 hour at an intensity of 80% of maximal work capacity. In this work protocol there are inserted 3 intervals of 5 minutes duration where the study subject works at 100% of maximal intensity with the purpose of securing a full activation/recruiting of all muscle fibers. Immediately following conclusion of the exercise bout 13C6-phenylalanine is infused for later determination of muscle protein synthesis. 2 hours into the recovery from exercise D2-glucose is infused for later determination of hepatic glucose production. 4 hours into the recovery from exercise an euglycaemic hyperinsulinemic clamp will be initiated for 2 hours for determination of insulin stimulated muscle glucose uptake. Muscle biopsies from the m. vastus lateralis muscle in one or both legs will be taken before, immediately after, as well as 2, 4 and 6 hours after conclusion of exercise. Pulmonary oxygen uptake is measured with the use of an online system before, during and after exercise and is used for determination of substrate metabolism. Blood samples from the femoral veins in both legs and the femoral artery in one of the legs are drawn before, during exercise, and during the recovery period after exercise. At the same time blood flow is measured in the femoral arteries in both legs with the use of Doppler technique. Determination of arteriovenous difference (AV difference) by simultaneous measurement of blood flow enables us to calculate skeletal muscle glucose uptake and uptake/release of relevant substances (proteins, peptides and metabolites etc.). The experimental part of trial A is completed at 15:30 p.m. The study subject will be given food and drink and will be observed for one hour before they may leave the research laboratory. Trial B is an identical protocol, except that the subject will receive opposite drug from trial A (Rapamycin or placebo). Trial A and B will be separated by a minimum of 14 days.

Official Title
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Mechanistic Target of Rapamycin (mTOR) as Mediator of Exercise-induced Insulin Sensitivity Study

Conditions
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Insulin Sensitivity

Intervention / Treatment
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* Other: Exercise
* Procedure: Insulin infusion
* Drug: Placebo
* Drug: Rapamycin


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy persons (no known disease) without diabetes in the family No use of medications Non-smokers Men Age 22-35 years Physical activity level (VO2peak) between 30-50 mL O2/min/kg BMI between 18.0 and 25.0 Exclusion Criteria: Women Physical activity (e.g., running, cycling, fitness, etc.) above 6 hours a week. Persons who show signs of metabolic diseases, hematologic diseases, reduced liver function, human immunodeficiency virus (HIV), hepatitis, thyroid diseases or other signs of disease that may affect the outcome of the study or the study subject's wellbeing will be excluded. Acute sickness less than 2 weeks prior to study start. If disease appears or is identified and/or a need for medication arises after inclusion but before initiation of the study. Disease during conclusion of the study

Ages Eligible for Study
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Minimum Age: 22 Years
Maximum Age: 35 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: Non-Randomized
Intervention Model: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Post-exercise insulin sensitivity following placebo administration<br>Young healthy males will ingest placebo (blinded) and perform a single bout of knee-extensor exercise and insulin action is investigated 4 hours after cessation of exercise. Insulin action towards muscle glucose uptake and protein synthesis will be investigated during a 120 min euglycemic hyperinsulinemic clamp. | Other: Exercise<br>* 1 hour of one-legged knee-extensor exercise followed by 4 hours of rest<br>Procedure: Insulin infusion<br>* 2 hours of Euglycemic Hyperinsulinemic Clamp<br>Drug: Placebo<br>* Administration of 800 mg of calcium tablets<br>|
| Experimental: Post-exercise insulin sensitivity following Rapamycin administration<br>Young healthy males will ingest Rapamycin (blinded) and perform a single bout of knee-extensor exercise and insulin action is investigated 4 hours after cessation of exercise. Insulin action towards muscle glucose uptake and protein synthesis will be investigated during a 120 min euglycemic hyperinsulinemic clamp. | Other: Exercise<br>* 1 hour of one-legged knee-extensor exercise followed by 4 hours of rest<br>Procedure: Insulin infusion<br>* 2 hours of Euglycemic Hyperinsulinemic Clamp<br>Drug: Rapamycin<br>* Administration of 16 mg of Rapamycin tablets<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Insulin stimulated muscle glucose uptake | Muscle glucose uptake during insulin stimulation in recovery from exercise (insulin sensitivity) will be determined based on Fick's principle. Thus, femoral arterio-venous difference (av difference) in blood glucose concentration will be multiplied by femoral arterial leg blood flow. | Through study completion, approx. 1 year. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Insulin stimulated muscle protein synthesis | Muscle protein synthesis during insulin stimulation in recovery from exercise will be determined based on incorporation of 13C6-phenylalanine in muscle biopsies. Unit of measure is fractional synthesis rate (FSR) in percentage per hour (%/h). | 1-2 year. | 
| Posttranslational modification of proteins in muscle biopsies | The obtained muscle samples will be subjected to mass spectrometry-based phosphoproteomic analysis. This measurement shows posttranslational modification that regulates protein function and will reveal the signalling network underlying the interventions studied. Unit of measure is log2-transformed arbitrary units. | 5 years | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Exercise, Muscle insulin Sensitivity, Muscle Glucose uptake, Muscle protein synthesis, mTOR