Mfn2 is critical for brown adipose tissue thermogenic function

Mitochondrial fusion and fission events, collectively known as mitochondrial dynamics, act as quality control mechanisms to ensure mitochondrial function and fine-tune cellular bioenergetics. Defective mitofusin 2 (Mfn2) expression and enhanced mitochondrial fission in skeletal muscle are hallmarks of insulin-resistant states. Interestingly, Mfn2 is highly expressed in brown adipose tissue (BAT), yet its role remains unexplored. Using adipose-specific Mfn2 knockout (Mfn2-adKO) mice, we demonstrate that Mfn2, but not Mfn1, deficiency in BAT leads to a profound BAT dysfunction, associated with impaired respiratory capacity and a blunted response to adrenergic stimuli. Importantly, Mfn2 directly interacts with perilipin 1, facilitating the interaction between the mitochondria and the lipid droplet in response to adrenergic stimulation. Surprisingly, Mfn2-adKO mice were protected from high-fat diet-induced insulin resistance and hepatic steatosis. Altogether, these results demonstrate that Mfn2 is a mediator of mitochondria to lipid droplet interactions, influencing lipolytic processes and whole-body energy homeostasis.

Effective treatment of mitochondrial myopathy by nicotinamide riboside, a vitamin B3

Johan Auwerx, Eija Riitta Pirinen

Nutrient availability is the major regulator of life and reproduction, and a complex cellular signaling network has evolved to adapt organisms to fasting. These sensor pathways monitor cellular energy metabolism, especially mitochondrial ATP production and NAD+/NADH ratio, as major signals for nutritional state. We hypothesized that these signals would be modified by mitochondrial respiratory chain disease, because of inefficient NADH utilization and ATP production. Oral administration of nicotinamide riboside (NR), a vitamin B3 and NAD+ precursor, was previously shown to boost NAD+ levels in mice and to induce mitochondrial biogenesis. Here, we treated mitochondrial myopathy mice with NR. This vitamin effectively delayed early- and late-stage disease progression, by robustly inducing mitochondrial biogenesis in skeletal muscle and brown adipose tissue, preventing mitochondrial ultrastructure abnormalities and mtDNA deletion formation. NR further stimulated mitochondrial unfolded protein response, suggesting its protective role in mitochondrial disease. These results indicate that NR and strategies boosting NAD+ levels are a promising treatment strategy for mitochondrial myopathy.

Wiley Open Access2014

Thermal microsensors for in vitro and in vivo monitoring of chemical and biological processes

Rima Padovani

Metabolism is a highly coordinated biochemical cellular activity indispensable for sustaining life, and is often related to mitochondrial functioning. Innovative technological approaches allowing the study of the thermal signature of chemical and metabolic processes are presented in this thesis. In particular, two solutions are proposed: (i) a nanocalorimetric platform enabling measurements of biochemical heat production in vitro, and (ii) a method for localized in vivo temperature measurements in mice using implantable miniaturized sensors. First, the design and development of the nanocalorimetric platform is presented. In particular, this platform is built around a commercial thermopile-based sensor chip, on which an open-well reservoir holding a sample with a volume of a few tens of microliter is directly positioned. Both components are embedded into an isothermal housing providing an excellent temperature stability (± 1 mK), which is a prerequisite for accurate heat power measurements. Furthermore, the platform is characterized by a fast thermalization time to the target temperature (less than 30 minutes) and a fast sensing response time (a few seconds). Numerical simulations, which allowed the optimization of the platform design, are also reported. Electrical calibration of the platform was carried out using resistive heaters positioned either on the sensor chip or in the sample reservoir. A heat power limit of detection of 70 nW was estimated. Then, two reactions for the validation of the nanocalorimetric platform were investigated: (i) the mixing of 1-propanol in water, and (ii) the oxidation of glucose catalyzed by glucose oxidase. The results showed good agreement with literature, confirming that our versatile platform may be applied to many thermochemical studies, including thermodynamic analysis and kinetic reaction analysis. Furthermore, localized in vivo temperature measurements in mice by means of implantable miniaturized sensors are presented. The aim was to monitor mouse metabolism during cold exposure, and to record possible temperature differences between the body temperature measured in the abdomen and the temperature of the brown adipose tissue (BAT) situated in the interscapular area. This approach is of biological interest as it may help unravelling the question whether biochemical activation of BAT is associated with local increase in metabolic heat production. Specifically, the preparation and calibration of miniaturized thermistor sensors are reported and the surgical procedure is also described. Temperature measurements, carried out on mice during cold exposure (6 °C) for a maximum duration of 6 hours, are presented and analyzed. Control measurements with a conventional probe confirmed the good performance of the implanted sensors. Moreover, analysis of the experimental results allowed the identification of two different mouse phenotypes, distinguishable in terms of their metabolic resistance to cold exposure. This difference was investigated from the thermal point of view by computational simulations. A simple physical model of the mouse body allowed to reproduce the global evolution of hypothermia and to explain qualitatively the temperature difference between abdomen and BAT locations. While with this approach, we have demonstrated the importance and feasibility of localized temperature measurements on mice, further optimization of this technique is foreseen to better identify local metabolism variations.

EPFL2016

Growth differentiation factor 15 is a myomitokine governing systemic energy homeostasis

Johan Auwerx, Dongryeol Ryu

Reduced mitochondria! electron transport chain activity promotes longevity and improves energy homeostasis via cell-autonomous and non-autonomous factors in multiple model systems. This mitohormetic effect is thought to involve the mitochondrial unfolded protein response (UPRmt), an adaptive stress-response pathway activated by mitochondria' proteotoxic stress. Using mice with skeletal muscle specific deficiency of Crifl (muscle-specific knockout [MKO]), an integral protein of the large mitoribosomal subunit (39S), we identified growth differentiation factor 15 (GDF15) as a UPRmt-associated cell non-autonomous myomitokine that regulates systemic energy homeostasis. MKO mice were protected against obesity and sensitized to insulin, an effect associated with elevated GDF15 secretion after UPRmt activation. In ob/ob mice, administration of recombinant GDF15 decreased body weight and improved insulin sensitivity, which was attributed to elevated oxidative metabolism and lipid mobilization in the liver, muscle, and adipose tissue. Thus, GDF15 is a potent mitohormetic signal that safeguards against the onset of obesity and insulin resistance.

Rockefeller Univ Press2017

Thermal microsensors for in vitro and in vivo monitoring of chemical and biological processes

Rima Padovani

EPFL2016