Vincent Voisin

Changes of phospholipid composition within the dystrophic muscle by matrix-assisted laser desorption/ionization mass spectrometry and mass spectrometry imaging.

Duchenne muscular dystrophy (DMD) is a neuromuscular disease linked to the lack of dystrophin, a submembrane protein, leading to muscle weakness and associated with a defect of the lipid metabolism. A study of the fatty acid composition of glycerophos-phatidylcholines by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) and tandem mass spectrometry (MS/MS) enabled us to characterize a change in the lipid composition of dystrophic cells at the time of the differentiation. This modification has been used as a marker to identify with profiling and imaging MALDI-ToF MS regenerating areas in sections of an mdx mouse leg muscle. It is the first time that such a slight change in fatty acid composition has been observed directly on tissue slices using mass spectrometry. This approach will be useful in monitoring the treatment of muscular regeneration.

l-arginine improves dystrophic phenotype in mdx mice

A possible treatment for Duchenne muscular dystrophies would be to compensate for dystrophin loss by increasing the expression of utrophin, another cytoskeletal protein of the muscle membrane. We previously found that L-arginine, the substrate for nitric oxide synthase, significantly increased utrophin level in muscle and targeted it to the sarcolemma. Here, we have addressed the expected benefit in the mdx mice. Magnetic resonance imaging of lower limbs revealed a 35% reduction of the necrotic zones, confirmed by histological staining of muscles. This regression of the necrosis was also supported by the drastic reduction of Evans blue incorporation, a cell impermeable dye. The creatine kinase level in the serum decreased by 57%. Utrophin level increased 2- to 3-fold in muscles. Beta-dystroglycan was relocalised with utrophin to the membrane. In the diaphragm, the most affected muscle in mdx mice, the isometric tension increased by 30%, with regression of collagen and of cytoplasmic lipid overloading. Finally, molsidomine, a therapeutic agent that is converted to a NO donor, also attenuated the dystrophic phenotype. Our results suggest that pharmacological activators of the NO pathway may constitute a realistic treatment for Duchenne and Becker muscular dystrophies.

Muscular nitric oxide synthase (muNOS) and utrophin

Duchenne muscular dystrophy (DMD), the severe X-linked recessive disorder which results in progressive muscle degeneration, is due to a lack of dystrophin, a membrane cytoskeletal protein. Three types of treatment are envisaged: pharmacological (glucocorticoid), myoblast transplantation, and gene therapy. An alternative to the pharmacological approach is to compensate for dystrophin loss by the upregulation of another cytoskeletal protein, utrophin. Utrophin and dystrophin are part of a complex of proteins and glycoproteins, which links the basal lamina to the cytoskeleton, thus ensuring the stability of the muscle membrane. One protein of the complex, syntrophin, is associated with a muscular isoform of the neuronal nitric oxide synthase (nNOS). We have demonstrated an overexpression of utrophin, visualised by immunofluorescence and quantified by Western blotting, in normal myotubes and in mdx (the animal model of DMD) myotubes, as in normal (C57) and mdx mice, both treated with nitric oxide (NO) donor or L-arginine, the NOS substrate. There is evidence that utrophin may be capable of performing the same cellular functions as dystrophin and may functionally compensate for its lack. Thus, we propose to use NO donors, as palliative treatment of Duchenne and Becker muscular dystrophies, pending, or in combination with, gene and/or cellular therapy. Discussion has focussed on the various isoforms of NOS that could be implicated in the regeneration process. Dystrophic and healthy muscles respond to treatment, suggesting that although NOS is delocalised in the cytoplasm in the case of DMD, it conserves substantial activity. eNOS present in mitochondria and iNOS present in cytoplasm and the neuromuscular junction could also be activated. Lastly, production of NO by endothelial NOS of the capillaries would also be beneficial through increased supply of metabolites and oxygen to the muscles.

Therapeutic Strategies for Duchenne and Becker Dystrophies

Duchenne muscular dystrophy (DMD), a severe X-linked genetic disease affecting one in 3500 boys, is the most common myopathy in children. DMD is due to a lack of dystrophin, a submembrane protein of the cytoskeleton, which leads to the progressive degeneration of skeletal, cardiac, and smooth muscle tissue. A milder form of the disease, Becker muscular dystrophy (BMD), is characterized by the presence of a semifunctional truncated dystrophin, or reduced levels of full-length dystrophin. DMD is the focus of three different supportive or therapeutic approaches: gene therapy, cell therapy, and drug therapy. Here we consider these approaches in terms of three potential goals: improvement of dystrophic phenotype, expression of dystrophin, and overexpression of utrophin. Utrophin exhibits 80% homology with dystrophin and is able to perform similar functions. Pharmacological strategies designed to overexpress utrophin appear promising and may circumvent many obstacles to gene and cell-based therapies.

Arginine butyrate: a therapeutic candidate for Duchenne muscular dystrophy

As a strategy to treat Duchenne muscular dystrophy, we used arginine butyrate, which combines two pharmacological activities: nitric oxide pathway activation, and histone deacetylase inhibition. Continuous intraperitoneal administration to dystrophin‐deficient mdx mice resulted in a near 2‐fold increase in utrophin (protein homologous to dystrophin) in skeletal muscle, heart, and brain, accompanied by an improvement of the dystrophic phenotype in both adult and newborn mice (45 and 70% decrease in creatine kinase level, respectively; 14% increase in tidal volume, 30% decrease in necrotic area in limb and 23% increase in isometric force). Intermittent administration, as performed in clinical trials, was then used to reduce the frequency of injections and to improve safety. This also enhanced utrophin level around 2‐fold (EC50=284 mg/ml) and alleviated the dystrophic phenotype (inverted grid and grip test performance near to wild‐type values, creatine kinase level decreased by 50%). Skin biopsies were used to monitor treatment efficacy, instead of invasive muscle biopsies, and this could be done a few days after the start of treatment. A 2‐fold increase in utrophin expression was also shown in cultured human myotubes. In vivo and in vitro experiments demonstrated that the drug combination acts synergistically. Together, these data constitute a proof of principle of the beneficial effects of arginine butyrate on muscular dystrophy.—Vianello, S., Yu, H., Voisin, V., Haddad, H., He, X., Foutz, A. S., Sebrié, C., Gillet, B., Roulot, M., Fougerousse, F., Perronnet, C., Vaillend, C., Matecki, S., Escolar, D., Bossi, L., Israël, M., de la Porte, S. Arginine butyrate: a therapeutic candidate for Duchenne muscular dystrophy. FASEB J. 27, 2256–2269 (2013). www.fasebj.org

Heterogeneous computing system platform for high-performance pattern recognition applications

we present a system architecture made of a motherboard with a Xilinx Zynq System on Chip (SoC) and a mezzanine board equipped with an Associative Memory chip (AM). The proposed architecture is designed to serve as an accelerator of general purpose algorithms based on pipeline processing and pattern recognition. We present the open source software and firmware developed to fully exploit the available communication channels between the ARM CPU and the FPGA using Direct Memory Access (DMA) technique and the AM using Multi-Gigabit Transceivers (MGT). We report the measured performances and discuss potential applications and future developments. The proposed architecture is compact, portable and provide a large communication bandwidth between components.