MedPath

Skeletal Muscle Mitochondria in Ageing

Recruiting
Conditions
Aging
Interventions
Other: Maximal exercise test
Other: 3D Ultrasound
Other: Dynamometry
Other: Exercise test and occlusions
Other: Exercise test in MRI
Procedure: Muscle biopsy
Registration Number
NCT06539078
Lead Sponsor
VU University of Amsterdam
Brief Summary

Healthy ageing is associated with the loss of muscle mass and physical function. As a result, older people are limited in their independence. The aging of muscles typically begins around the age of 30. From this age onward, muscle strength, muscle mass, and the maximum oxygen uptake of muscles decrease. The reasons for this are not entirely clear, but it seems to be partly related to how oxygen moves from our blood vessels to the muscles and how muscles burn energy. The precise role of age and physical fitness, as well as whether exercise can counteract the effects of ageing, is still unknown. Therefore, in this study, we aim to investigate the muscle function of both physically active and inactive young and middle-aged individuals. We hypothesise that endurance training can mitigate some of the effects of ageing.

Detailed Description

Healthy ageing is associated with a loss of muscle mass and physical function. This loss of physical function is underpinned by reductions in characteristics such as muscle strength, power, and maximal oxygen uptake (V̇O2max; reflecting exercise capacity). However, the causal contributors to these age-associated impairments, and the role of exercise training status in mitigating them, remain poorly defined. Skeletal muscle mitochondrial function has been proposed to be a key contributor to age-associated effects on physical function, however many conflicting results are present in the extant human literature. Moreover, diffusion of oxygen from capillaries to mitochondria is a key determinant of V̇O2max, however, whether the skeletal muscle diffusive capacity for oxygen (DmO2) declines with age is unknown. A new technique utilizing near-infrared spectroscopy (NIRS) will enable the non-invasive assessment of skeletal muscle diffusive capacity in young and elderly subjects for the first time to resolve this issue. The primary aims of this study are therefore to 1) compare DmO2 derived via NIRS between young sedentary, young endurance-trained, older sedentary, and older endurance-trained subjects; 2) to compare non-invasive (i.e. with NIRS and 31phosphorous magnetic resonance spectroscopy \[31P-MRS\]) and invasive (i.e. measures of mitochondrial morphology and respiration obtained by skeletal muscle biopsy) markers of mitochondrial function between the same groups, and 3) to assess the relationships between DmO2, mitochondrial measures and assessments of capillarization with functional measurements of muscle strength, power, and V̇O2max.

Recruitment & Eligibility

Status
RECRUITING
Sex
All
Target Recruitment
60
Inclusion Criteria

In order to be eligible to participate in this study, young sedentary participants must meet all of the following criteria:

  • Aged between 18-30 years
  • Male or female
  • Not currently engaging in any formal exercise training or competitive sports
  • No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise

In order to be eligible to participate in this study, young trained participants must meet all of the following criteria:

  • Aged between 18-30 years
  • Male or female
  • Currently engaging in formal training (at least 3 times per week) in competitive endurance sports
  • No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise

In order to be eligible to participate in this study, older sedentary participants must meet all of the following criteria:

  • Aged between 50-65 years
  • Male or female
  • Not currently engaging in any formal exercise training or competitive sports
  • No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise

In order to be eligible to participate in this study, older trained participants must meet all of the following criteria:

  • Aged between 50-65 years
  • Male or female
  • Currently engaging in formal training (at least 3 times per week) in competitive endurance sports
  • No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Exclusion Criteria
  • Age that falls outside of 18-30 years (young groups) or 50-65 years (middle-aged groups)
  • Inability to provide informed consent
  • History of claustrophobia
  • Ineligibility to perform the exercise test described in this study protocol or follow instructions
  • Taking any medications known to interfere with the physiological responses to exercise, e.g. e.g. systemic corticosteroids, statins, SGLT2 inhibitors, GLP1 receptor agonists
  • Contraindication for MRI (e.g. pacemaker, claustrophobia)
  • Being under investigation for non-diagnosed disease at the time of investigation
  • Body Mass Index (BMI) >30 due to adiposity, since this is known to cause difficulties in obtaining muscle biopsies and NIRS measurements
  • Pregnancy
  • Are current smokers or have been a regular smoker within the last 12 months

Study & Design

Study Type
OBSERVATIONAL
Study Design
Not specified
Arm && Interventions
GroupInterventionDescription
Young, sedentary participants3D Ultrasound* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participantsMaximal exercise test* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participantsMuscle biopsy* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participantsExercise test in MRI* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participantsMuscle biopsy* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participants3D Ultrasound* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young, sedentary participantsMaximal exercise test* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participantsMaximal exercise test* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participantsDynamometry* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participantsMaximal exercise test* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participantsExercise test in MRI* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young, sedentary participantsExercise test and occlusions* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participantsExercise test and occlusions* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participantsExercise test and occlusions* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young, sedentary participantsDynamometry* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participants3D Ultrasound* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participantsExercise test in MRI* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young, sedentary participantsExercise test in MRI* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young, sedentary participantsMuscle biopsy* Aged between 18-30 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participantsDynamometry* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Young endurance-trained participantsExercise test and occlusions* Aged between 18-30 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged, sedentary participants3D Ultrasound* Aged between 50-65 years * Male or female * Not currently engaging in any formal exercise training or competitive sports * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participantsDynamometry* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Middle-aged endurance-trained participantsMuscle biopsy* Aged between 50-65 years * Male or female * Currently engaging in formal training (at least 3 times per week) in competitive endurance sports (i.e. running, cycling, swimming, triathlon, etc.) * No chronic health conditions likely to affect exercise tolerance or the physiological responses to exercise
Primary Outcome Measures
NameTimeMethod
Muscle strengthBaseline (visit 1)

Newton-metre (Nm)

Muscle powerBaseline (visit 1)

Watt (W)

Muscle mitochondrial fragmentation index (A.U.)Visit 6 muscle biopsy (+/- after 4 weeks).

Degree of fragmentation of the mitochondrial pool.

Maximal oxygen uptake (V̇O2max)Baseline (visit 1)

ml/min/kg

Muscle volumeBaseline (visit 1)

cm\^3

Muscle diffusing capacity for oxygen (DmO2)Baseline (visit 1) and visit 2-4. In total 4 weeks.

Differences in recovery constant k (min-1) obtained under conditions of high, medium or low O2 availability

Secondary Outcome Measures
NameTimeMethod
(Peak) power outputBaseline (visit 1)

Watt (W)

Capillary lactate concentrationBaseline (visit 1)

mmol/L

NIRS derived muscle oxy- and deoxygenated [haemoglobin + myoglobin] (HbO2, Hbb) and tissue saturation index (TSI) versus relative and absolute work rate.Baseline (visit 1)

HbO2 and Hbb: % maximal value, TSI (%) = HbO2/(HbO2+Hbb) versus relative (%max) and absolute power output (W) Relative and absolute work rates comparisons will be reported for all variables: resting concentration, baseline cycling concentration, (sub)maximal exercise concentration.

Initial and secondary slope of increase during incremental exercise will be reported for NIRS derived muscle oxy- and deoxygenated [haemoglobin + myoglobin] (HbO2, Hbb) and tissue saturation index (TSI) versus relative work rate.Baseline (visit 1)

concentration\[Hbb/HbO2/TSI\]%/delta%peak power(W)

Maximal ventilatory equivalentsBaseline (visit 1)

VE/VCO2 and VE/VO2

Pulmonary oxygen uptake - baseline and steady state V̇O2Baseline (visit 1) and visit 2-4 (max 4 weeks in total)

L/min

Pulmonary oxygen uptake kinetics - Phase II V̇O2 time constant and time delayVisit 2-4 (max 4 weeks in total)

sec

Muscle (de)oxygenation breakpoint during incremental exerciseBaseline (visit 1)

Power output (W) and maximal oxygen uptake (L/min)

Muscle morphology - pennation angleBaseline (visit 1)

degrees

Mean and fiber type specific succinate dehydrogenase activityVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

A660/um/s

Mitochondrial surface area-to-volume ratioVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

um/um\^2

Mean response time of the V̇O2 slope during ramp exerciseBaseline (visit 1)

sec

V̇O2/HR slope during ramp exerciseBaseline (visit 1)

beats/L/min

Maximal respiratory exchange ratio (RER)Baseline (visit 1)

RER = VCO2/VO2

Maximal end-tidal pressures for oxygen (O2) and carbon dioxide (CO2)Baseline (visit 1)

mmHg

Rate constant of mV̇O2 recovery kinetics under conditions of high, medium and low O2 availabilityBaseline (visit 1)

sec

Respiratory control ratiosVisit 6 muscle biopsy (+/- after 4 weeks).

OXPHOS/ETS, LEAK/ETS, LEAK/OXPHOS, LEAK/NADH-linked, ROT+S/ETS, (OXPHOS-LEAK)/ETS, (OXPHOS-LEAK)/OXPHOS, (ETS-LEAK)/ETS, (ETS-OXPHOS)/ETS

Intrinsic mitochondrial respiration (each respiratory state outlined below will be normalised to mitochondrial volume density)Visit 6 muscle biopsy (+/- after 4 weeks).

(pmol/s/mg)/mitochondrial area density (%) For: background, LEAK, N-linked respiration, OXPHOS, ETS, succinate + rotenone-linked uncoupled respiration

31P-MRS-derived mitochondrial bioenergetic function - resting and steady-state exercising concentrations and amplitude of exercise-induced changes of skeletal muscle Phosphocreatine [PCr], [inorganic phosphate] and pHVisit 5 MRI

mM pH = unitless

31P-MRS-derived mitochondrial bioenergetic function - maximal rate of oxidative ATP synthesisVisit 5 MRI

mM/s

(mean) fiber cross-sectional area, also fiber type specificVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

um

Mean and fiber type specific myoglobin concentrationsVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

mM

Muscle capillarization - capillary densityVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

number of capillaries/mm2

Muscle capillarization - capillary-to-fiber ratioVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

capillary-to-fiber ratio

Muscle capillarization - length of capillaries relative to fiber perimeter (LC/PF)Visit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

percentage (%)

Mitochondrial area densityVisit 6 muscle biopsy (+/- after 4 weeks) - electron microscopy (EM) within time window of 2 years.

Percentage (%)

Mitochondrial numberVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

number/um2

Gas exchange and ventilatory variables (gas exchange threshold, respiratory compensation point, maximal ventilation)Baseline (visit 1)

L/min

(Maximum) heart rate (HR)Baseline (visit 1) and during visit 2-4 (max 4 weeks in total)

beats per minute (bpm)

Pulmonary oxygen uptake kinetics - Phase II V̇O2 amplitudeVisit 2-4 (max 4 weeks in total)

L/min

Concentrations of NIRS derived muscle oxy- and deoxygenated [haemoglobin + myoglobin] (HbO2, Hbb) and tissue saturation index (TSI).Baseline (visit 1) and visit 2-4 (max 4 weeks in total)

HbO2 and Hbb: % maximal value, TSI (%) = HbO2/(HbO2+Hbb) For all variables resting concentration, baseline cycling concentration, (sub)maximal exercise concentration will be reported.

Initial and secondary slope during incremental exercise will be reported for NIRS derived muscle oxy- and deoxygenated [haemoglobin + myoglobin] (HbO2, Hbb) and tissue saturation index (TSI) versus absolute work rate.Baseline (visit 1)

concentration\[Hbb/HbO2/TSI\]%/deltaW

Muscle morphology - (effective) physiological cross-sectional area (PCSA)Baseline (visit 1)

cm2

Adipose tissue thickness at the site of NIRS measurementBaseline (visit 1)

mm

Maximal O2 pulseBaseline (visit 1)

ml/beat

Slope of ventilation (VE) versus carbon dioxide (VCO2) output during ramp exercise (i.e. ventilatory efficiency)Baseline (visit 1)

V̇E/V̇CO2 slope

Maximal respiratory frequencyBaseline (visit 1)

breaths/min

Mitochondrial respiratory function (background, LEAK, N-linked respiration, OXPHOS, ETS, succinate (S) + rotenone (ROT)-linked uncoupled respiration, )Visit 6 muscle biopsy (+/- after 4 weeks).

pmol/s/mg

31P-MRS-derived mitochondrial bioenergetic function rate constant of PCrVisit 5 MRI

Rate constant of Phosphocreatine \[PCr\] on- and off-kinetics (sec)

Weighted fiber cross-sectional areaVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

um2

Muscle capillarization - mean number of capillaries surrounding a fiber (CAF)Visit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

number of capillaries

Muscle capillarization - mean number of capillaries surrounding a fiber in relation to fiber area (CAFA)Visit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

number of capillaries/um2

Muscle capillarization - capillary-to fiber-perimeter exchange index (CFPE)Visit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

CFPE (unitless)

Muscle capillarization - sarcomere lengthVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

um

Mitochondrial circularityVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

Circularity (AU)

Mitochondrial cristae area densityVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

Percentage (%)

Muscle morphology - vastus lateralis specific forceBaseline (visit 1)

N/cm2

Muscle morphology - estimated muscle fiber numberBaseline (visit 1)

PCSA/muscle fiber cross-sectional area

Muscle fiber type distributionVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

Type I, IIa, IIx and hybrid fiber-type proportions (%)

Integrated fiber succinate dehydrogenase activityVisit 6 muscle biopsy (+/- after 4 weeks) - staining within time window of 2 years.

A660.um/s

Mitochondrial dynamics proteins (Mfn1, Mfn2, OPA1, Drp1, Parkin, PINK1, Fis1, MTFP1, NRF1&2, PGC1a, TFAM, OXPHOS protein content (complexes I-V and total protein content)Visit 6 muscle biopsy (+/- after 4 weeks) - western blot within time window of 2 years.

ug/mg loaded sample

Physical activity status - pedometerBaseline 7 days

Number of steps per day

Mitochondrial areaVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

um2

Mitochondrial height, width, perimeter and maximal+minimal Feret's diameterVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

um

Mitochondrial volume densityVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

um3.um3.10\^2

Mitochondrial roundnessVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

Roundness (AU)

Mitochondrial aspect ratioVisit 6 muscle biopsy (+/- after 4 weeks) - EM within time window of 2 years.

Aspect ratio (AU)

Muscle morphology - Fascicle lengthBaseline (visit 1)

cm

Trial Locations

Locations (1)

Vrije Universiteit Amsterdam

🇳🇱

Amsterdam, Noord-Holland, Netherlands

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