The Interactome in the Evolution From Frailty to Sarcopenic Dependence

Biomarkers are essential tools for accurate diagnosis and effective prevention, but their validation is a pending challenge that limits their usefulness, even more so with constructs as complex as frailty. Sarcopenia shares multiple mechanisms with frailty which makes it a strong candidate to provide robust frailty biomarkers. Based on this premise, we studied the temporal evolution of cellular interactome in frailty, from independent patients to dependent ones. Overweight is a recognized cause of frailty in aging, so we studied the altered mechanisms in overweight independent elderly and evaluated their aggravation in dependent elderly. This evidence of the evolution of previously altered mechanisms would significantly support their role as real biomarkers of frailty. The results showed a preponderant role of autophagy in interactome control at both different functional points, modulating other essential mechanisms in the cell, such as mitochondrial capacity or oxidative stress. Thus, the overweight provoked in the muscle of the elderly an overload of autophagy that kept cell survival in apparently healthy individuals. This excessive and permanent autophagic effort did not seem to be able to be maintained over time. Indeed, in dependent elderly, the muscle showed a total autophagic inactivity, with devastating effects on the survival of the cell, which showed clear signs of apoptosis, and reduced functional capacity. The frail elderly are in a situation of weakness that is a precursor of dependence that can still be prevented if detection is early. Hence biomarkers are essential in this context.

Initial Results From SOMMA: Contribution of Mitochondrial Function to Walking and Fitness

We hypothesize that the capacity of mitochondria in quadriceps skeletal muscle to generate ATP energy by respirometry (OXPHOS) in biopsies from the vastus lateralis, and in whole quadriceps muscle by 31PMRS (ATPmax) would contribute to 4 and 400m gait speed and to peak oxygen consumption on treadmill testing (VO2peak). In analyses from the first SOMMA participants recruited (N=122), OXPHOS was similarly associated with 4m (r=0.21) and 400 m (r=0.21) walking speed (P<0.01). However, ATPmax was not associated with either 4m or 400m walking speed (r=-0.02 and -0.07 respectively). In contrast both OXPHOS (r=0.43) and ATP max (r=0.35) were more strongly correlated with fitness (VO2 peak). These findings suggest that in older people, the mitochondrial capacity to generate ATP plays an important role walking speed and may be even more important to fitness.

Dark respiration rates are not determined by differences in mitochondrial capacity, abundance and ultrastructure in C4 leaves

Our understanding of the regulation of respiration in C plants, where mitochondria play different roles in the different types of C photosynthetic pathway, remains limited. We examined how leaf dark respiration rates (R), in the presence and absence of added malate, vary in monocots representing the three classical biochemical types of C photosynthesis (NADP-ME, NAD-ME and PCK) using intact leaves and extracted bundle sheath strands. In particular, we explored to what extent R are associated with mitochondrial number, volume and ultrastructure. We found that the respiratory response of NAD-ME and PCK type bundle sheath strands to added malate was associated with differences in mitochondrial number, volume, and/or ultrastructure, while NADP-ME type bundle sheath strands did not respond to malate addition. In general, mitochondrial traits reflected the contributions mitochondria make to photosynthesis in the three C types. However, despite the obvious differences in mitochondrial traits, no clear correlation was observed between these traits and R. We suggest that R is primarily driven by cellular maintenance demands and not mitochondrial composition per se, in a manner that is somewhat independent of mitochondrial organic acid cycling in the light.

Butyrate Alters Pyruvate Flux and Induces Lipid Accumulation in Cultured Colonocytes

Butyrate is considered the primary energy source of colonocytes and has received wide attention due to its unique health benefits. Insight into the mechanistic effects of butyrate on cellular and metabolic function relies mainly on research in in-vitro-cultured cells. However, cells in culture differ from those in vivo in terms of metabolic phenotype and nutrient availability. For translation, it is therefore important to understand the impact of different nutrients on the effects of butyrate. We investigated the metabolic consequences of butyrate exposure under various culturing conditions, with a focus on the interaction between butyrate and glucose. To investigate whether the effects of butyrate were different between cell­­s with high and low mitochondrial capacity, we cultured HT29 cells under either low- (0.5 mM) or high- (25 mM) glucose conditions. Low-glucose culturing increased the mitochondrial capacity of HT29 cells compared to high-glucose (25 mM) cultured HT29 cells. Long-term exposure to butyrate did not alter mitochondrial bioenergetics, but it decreased glycolytic function, regardless of glucose availability. In addition, both high- and low-glucose-grown HT29 cells showed increased lipid droplet accumulation following long-term butyrate exposure. Acute exposure of cultured cells (HT29 and Caco-2) to butyrate increased their oxygen consumption rate (OCR). A simultaneous decrease in extracellular acidification rate (ECAR) was observed. Furthermore, in the absence of glucose, OCR did not increase in response to butyrate. These results lead us to believe that butyrate itself was not responsible for the observed increase in OCR, but, instead, butyrate stimulated pyruvate flux into mitochondria. Indeed, blocking of the mitochondrial pyruvate carrier prevented a butyrate-induced increase in oxygen consumption. Taken together, our results indicate that butyrate itself is not oxidized in cultured cells but instead alters pyruvate flux and induces lipid accumulation.

Impact of aging and exercise on skeletal muscle mitochondrial capacity, energy metabolism, and physical function

The relationship between the age-associated decline in mitochondrial function and its effect on skeletal muscle physiology and function remain unclear. In the current study, we examined to what extent physical activity contributes to the decline in mitochondrial function and muscle health during aging and compared mitochondrial function in young and older adults, with similar habitual physical activity levels. We also studied exercise-trained older adults and physically impaired older adults. Aging was associated with a decline in mitochondrial capacity, exercise capacity and efficiency, gait stability, muscle function, and insulin sensitivity, even when maintaining an adequate daily physical activity level. Our data also suggest that a further increase in physical activity level, achieved through regular exercise training, can largely negate the effects of aging. Finally, mitochondrial capacity correlated with exercise efficiency and insulin sensitivity. Together, our data support a link between mitochondrial function and age-associated deterioration of skeletal muscle.

In vivo mitochondrial ATP production is improved in older adult skeletal muscle after a single dose of elamipretide in a randomized trial

Background Loss of mitochondrial function contributes to fatigue, exercise intolerance and muscle weakness, and is a key factor in the disability that develops with age and a wide variety of chronic disorders. Here, we describe the impact of a first-in-class cardiolipin-binding compound that is targeted to mitochondria and improves oxidative phosphorylation capacity (Elamipretide, ELAM) in a randomized, double-blind, placebo-controlled clinical trial. Methods Non-invasive magnetic resonance and optical spectroscopy provided measures of mitochondrial capacity (ATPmax) with exercise and mitochondrial coupling (ATP supply per O2 uptake; P/O) at rest. The first dorsal interosseous (FDI) muscle was studied in 39 healthy older adult subjects (60 to 85 yrs of age; 46% female) who were enrolled based on the presence of poorly functioning mitochondria. We measured volitional fatigue resistance by force-time integral over repetitive muscle contractions. Results A single ELAM dose elevated mitochondrial energetic capacity in vivo relative to placebo (ΔATPmax; P = 0.055, %ΔATPmax; P = 0.045) immediately after a 2-hour infusion. No difference was found on day 7 after treatment, which is consistent with the half-life of ELAM in human blood. No significant changes were found in resting muscle mitochondrial coupling. Despite the increase in ATPmax there was no significant effect of treatment on fatigue resistance in the FDI. Conclusions These results highlight that ELAM rapidly and reversibly elevates mitochondrial capacity after a single dose. This response represents the first demonstration of a pharmacological intervention that can reverse mitochondrial dysfunction in vivo immediately after treatment in aging human muscle.

Dissection of two routes to naïve pluripotency using different kinase inhibitors

Embryonic stem cells (ESCs) can be maintained in the naïve state through inhibition of Mek1/2 and Gsk3 (2i). A relevant effect of 2i is the inhibition of Cdk8/19, which are negative regulators of the Mediator complex, responsible for the activity of enhancers. Inhibition of Cdk8/19 (Cdk8/19i) stimulates enhancers and, similar to 2i, stabilizes ESCs in the naïve state. Here, we use mass spectrometry to describe the molecular events (phosphoproteome, proteome, and metabolome) triggered by 2i and Cdk8/19i on ESCs. Our data reveal widespread commonalities between these two treatments, suggesting overlapping processes. We find that post-transcriptional de-repression by both 2i and Cdk8/19i might support the mitochondrial capacity of naive cells. However, proteome reprogramming in each treatment is achieved by different mechanisms. Cdk8/19i acts directly on the transcriptional machinery, activating key identity genes to promote the naïve program. In contrast, 2i stabilizes the naïve circuitry through, in part, de-phosphorylation of downstream transcriptional effectors.

Mitochondrial Capacity and Muscle Endurance in Individuals with Parkinson’s Disease

Introduction: Parkinson's disease (PD) is associated with loss of motor control and difficulty exercising. This study measured skeletal muscle mitochondrial capacity and endurance in individuals with and without PD using novel non-invasive methods. We hypothesized that individuals with PD will have decreased mitochondrial capacity, reduced oxygen recovery, and decreased endurance compared to controls. Methods: Eight participants with PD and nine healthy controls were tested. Mitochondrial capacity was measured as the rate of recovery of muscle metabolism after electrical stimulation using near-infrared spectroscopy (NIRS) and repeated short arterial occlusions. Oxygen recovery was measured as the half time of recovery of oxygen levels after 5 minutes of ischemia. Muscle endurance was determined from changes in twitch contraction acceleration during electrical stimulation at 2, 4, and 6 Hz. Results: Mitochondrial capacity was lower in individuals with PD compared to controls (1.5±0.1min-1 vs. 1.7±0.1min-1, p=0.02). Individuals with PD had slower oxygen recovery after ischemia compared to controls (8.9±2.3s vs. 5.4±0.8s, p=0.01). Endurance was not different between groups at 6 Hz (PD vs controls: 58±23% vs. 69±16%, p=0.34). The effect sizes for mitochondrial capacity and oxygen recovery were large (Cohen's d >0.8). The Cohen's d for endurance was 1.11. Conclusion: Individuals with PD had slight impairments in mitochondrial capacity and blood flow but did not have reduced muscle endurance. While our study suggests that muscle metabolic dysfunction may play a minor role in exercise intolerance in people with PD, it demonstrates the use of noninvasive technologies to evaluate muscle function in people with neurological disorders.